Erkki Savilahti

The increase in allergic diseases is attributed to a relative lack of microbial stimulation of the infantile gut immune system. Probiotics, live health-promoting microbes, might offer such stimulation.We studied the effect of a mixture of 4 probiotic bacterial strains along with prebiotic galacto-oligosaccharides in preventing allergic diseases.We randomized 1223 pregnant women carrying high-risk children to use a probiotic preparation or a placebo for 2 to 4 weeks before delivery. Their infants received the same probiotics plus galacto-oligosaccharides (n = 461) or a placebo (n = 464) for 6 months. At 2 years, we evaluated the cumulative incidence of allergic diseases (food allergy, eczema, asthma, and allergic rhinitis) and IgE sensitization (positive skin prick test response or serum antigen-specific IgE level >0.7 kU/L). Fecal bacteria were analyzed during treatment and at age 2 years.Probiotic treatment compared with placebo showed no effect on the cumulative incidence of allergic diseases but tended to reduce IgE-associated (atopic) diseases (odds ratio [OR], 0.71; 95% CI, 0.50-1.00; P = .052). Probiotic treatment reduced eczema (OR, 0.74; 95% CI, 0.55-0.98; P = .035) and atopic eczema (OR, 0.66; 95% CI, 0.46-0.95; P = .025). Lactobacilli and bifidobacteria more frequently (P < .001) colonized the guts of supplemented infants.Probiotic treatment showed no effect on the incidence of all allergic diseases by age 2 years but significantly prevented eczema and especially atopic eczema. The results suggest an inverse association between atopic diseases and colonization of the gut by probiotics.The prevention of atopic eczema in high-risk infants is possible by modulating the infant's gut microbiota with probiotics and prebiotics.

Tissue transglutaminase has been reported to be the target for endomysial antibodies in celiac disease. We sought to establish whether immunoglobulin (Ig) A class tissue transglutaminase autoantibodies can be considered specific for celiac disease.Serum samples from 136 patients with untreated celiac disease (diagnosed according to the criteria of the European Society for Pediatric Gastroenterology, Hepatology and Nutrition) and 207 disease controls were studied. Enzyme-linked immunosorbent assay (ELISA) and Western blots were performed using calcium-treated and untreated tissue transglutaminase as antigen. Reticulin, endomysial, and mouse monoclonal tissue transglutaminase antibodies were studied by an indirect immunofluorescence method and gliadin antibodies with ELISA.The calcium-activated tissue transglutaminase autoantibody ELISA was highly sensitive (129 of 136) and specific (194 of 207) in detecting celiac disease. The new autoantibody ELISA test correlated well with the endomysial antibody test. Tissue transglutaminase autoantibody ELISA showed a clearly better predictive potential than the IgA class gliadin antibody ELISA. Immunoblots and ELISA blocking studies showed that calcium is needed for the specific antigen-antibody reaction to occur. Double immunofluorescence staining in human umbilical cord with sera from patients with celiac disease and with monoclonal tissue transglutaminase antibodies showed complete overlap.Calcium-activated tissue transglutaminase autoantibody ELISA is highly accurate in detecting untreated celiac disease. Tissue transglutaminase seems to be the target self-antigen for endomysial antibodies.

Lactobacillus rhamnosus GG, ATCC (LGG), has shown antagonism to many bacteria including mutans streptococci. This randomized, double-blind, placebo-controlled intervention study was designed to examine whether milk containing LGG has an effect on caries and the risk of caries in children when compared with normal milk. 594 children, 1-6 years old, from 18 municipal day-care centres were included. The children received the milk with meals from coded containers 5 days a week in the day-care centres for 7 months. The children's oral health was recorded at baseline and at the end, using WHO criteria. The caries risk was calculated based on clinical and microbiological data, comprising mutans streptococcus levels from dental plaque and saliva. The risk was classified as high if the child had a dmft/DMFT or initial caries score >0, and a mutans streptococcus count > or = 10(5) CFU/ml. The results showed less dental caries in the LGG group and lower mutans streptococcus counts at the end of the study. LGG was found to reduce the risk of caries significantly (OR = 0.56, p = 0.01; controlled for age and gender, OR = 0.51, p = 0.004). The effect was particularly clear in the 3- to 4-year-olds. Thus, milk containing the probiotic LGG bacteria may have beneficial effects on children's dental health.

Probiotic bacteria are suggested to reduce symptoms of the atopic eczema/dermatitis syndrome (AEDS) in food-allergic infants. We aimed to investigate whether probiotic bacteria have any beneficial effect on AEDS.Follow-up of severity of AEDS by the Severity Scoring of Atopic Dermatitis (SCORAD) index in 230 infants with suspected cow's milk allergy (CMA) receiving, in a randomized double-blinded manner, concomitant with elimination diet and skin treatment, Lactobacillus GG (LGG), a mixture of four probiotic strains, or placebo for 4 weeks. Four weeks after the treatment, CMA was diagnosed with a double-blind placebo-controlled (DBPC) milk challenge in 120 infants.In the whole group, mean SCORAD (at baseline 32.5) decreased by 65%, but with no differences between treatment groups immediately or 4 weeks after the treatment. No treatment differences were observed in infants with CMA either. In IgE-sensitized infants, however, the LGG group showed a greater reduction in SCORAD than did the placebo group, -26.1 vs-19.8 (P=0.036), from baseline to 4 weeks after the treatment. Exclusion of infants who had received antibiotics during the study reinforced the findings in the IgE-sensitized subgroup.Treatment with LGG may alleviate AEDS symptoms in IgE-sensitized infants but not in non-IgE-sensitized infants.

Correlation of endoscopic Crohn's disease activity with fecal calprotectin and lactoferrin is insufficiently studied. We evaluated the clinical significance of these neutrofil-derived proteins in assessment of Crohn's disease activity by comparing them with endoscopic disease activity and with Crohn's disease activity index (CDAI) and serum CRP.A total of 77 CD patients underwent one or more ileocolonoscopies (n = 106) with scoring of Crohn's disease index of severity (CDEIS). Patients provided stool samples for calprotectin and lactoferrin measurements and blood samples for CRP. Clinical activity was based on the CDAI.Both fecal calprotectin and lactoferrin correlated significantly with CDEIS (Spearman's r 0.729 and 0.773, P < 0.001). With a cutoff level of 200 microg/g for a raised fecal calprotectin concentration, sensitivity was 70%, specificity 92%, positive predictive value (PPV) 94%, and negative predictive value (NPV) 61% in predicting endoscopically active disease (CDEIS >/= 3). A fecal lactoferrin concentration of 10 microg/g as the cutoff value gave a sensitivity, specificity, PPV, and NPV of 66%, 92%, 94%, and 59%. Sensitivity of CDAI >/= 150 to detect endoscopically active disease was only 27%, specificity 94%, PPV 91%, and NPV 40%. A raised serum CRP (> 5 mg/l) gave a sensitivity, specificity, PPV, and NPV of 48%, 91%, 91%, and 48%.For evaluation of Crohn's disease activity, based on endoscopic findings, more sensitive surrogate markers than is CDAI or CRP are fecal calprotectin and lactoferrin. These prove to be useful tools for estimation of disease activity in Crohn's disease.

<h3>Background</h3> Less microbial exposure in early childhood is associated with more allergic disease later. Allergic children have a different fecal microflora, with less lactobacilli and bifidobacteria. Beneficial effects regarding the development of allergy have been suggested to come through probiotic supplementation. <h3>Objective</h3> We sought to study the effect of probiotic and prebiotic supplementation in preventing allergies. <h3>Methods</h3> In a double-blinded, placebo-controlled study we randomized 1223 mothers with infants at high risk for allergy to receive a probiotic mixture (2 lactobacilli, bifidobacteria, and propionibacteria) or placebo during the last month of pregnancy and their infants to receive it from birth until age 6 months. Infants also received a prebiotic galacto-oligosaccharide or placebo. At 5 years, we evaluated the cumulative incidence of allergic diseases (eczema, food allergy, allergic rhinitis, and asthma) and IgE sensitization. <h3>Results</h3> Of the 1018 intent-to-treat infants, 891 (88%) attended the 5-year visit. Frequencies of allergic and IgE-associated allergic disease and sensitization in the probiotic and placebo groups were similar: 52.6% versus 54.9% and 29.5% versus 26.6%, respectively, and 41.3% in both. No significant difference appeared in frequencies of eczema (39.3% vs 43.3%), atopic eczema (24.0% vs 25.1%), allergic rhinitis (20.7% vs 19.1%), or asthma (13.0% vs 14.1%) between groups. However, less IgE-associated allergic disease occurred in cesarean-delivered children receiving probiotics (24.3% vs 40.5%; odds ratio, 0.47; 95% CI, 0.23% to 0.96%; <i>P</i> = .035). <h3>Conclusions</h3> No allergy-preventive effect that extended to age 5 years was achieved with perinatal supplementation of probiotic bacteria to high-risk mothers and children. It conferred protection only to cesarean-delivered children.

Probiotic bacteria are potentially beneficial to maturation of the infant's immune system.To examine the role of probiotic bacteria in treatment of cow's milk allergy (CMA) and IgE-associated dermatitis, we investigated the immunologic effects of Lactobacillus rhamnosus GG (LGG) and a mixture of 4 bacterial species (MIX).In a randomized, double-blind study design, concomitantly with elimination diet and skin treatment, LGG, MIX, or placebo was given for 4 weeks to infants with suspected CMA. After anti-CD3 (OKT3) and anti-CD28 stimulation of PBMCs, IFN-gamma, IL-4, IL-5, and IL-12 levels were measured in culture supernatants by ELISA. Intracellular IFN-gamma, IL-4, and IL-5 production on CD4 lymphocytes was analyzed with fluorescence-activated cell sorting.Secretion of IFN-gamma by PBMCs before the treatment was significantly lower in infants with CMA (P=.016) and in infants with IgE-associated CMA (P=.003) than in non-CMA infants. Among the infants who received LGG, the level of secreted IFN-gamma increased in those with CMA (P=.006) and in those with IgE-associated dermatitis (P=.017) when compared with the placebo group. Secretion of IL-4 increased significantly in infants with CMA in the MIX (P=.034) but not in the LGG group.Deficiency in IFN-gamma response appears to be related to CMA. LGG raises IFN-gamma production of PBMC in infants with CMA and in infants with IgE-associated dermatitis and may thus provide beneficial TH1 immunomodulatory signals. MIX, although containing LGG, appears to modulate the immune responses differently.

Faecal calprotectin and lactoferrin increasingly serve as surrogate markers of disease activity in IBD. Data on the correlation of these markers with simple endoscopic score for Crohn's disease (SES-CD) and with histological findings are as yet limited. Aim To study the correlation of faecal calprotectin and lactoferrin with SES-CD and histology.During 87 consecutive ileocolonoscopies, SES-CD was calculated and biopsy specimens were obtained from the ileum, colon and rectum. Faecal calprotectin and lactoferrin were measured.In ileocolonic or colonic disease, both faecal calprotectin and lactoferrin correlated significantly with colon SES-CD (P < 0.001) and colon histology (P < 0.001). In patients with normal calprotectin or lactoferrin levels, endoscopic and histology scores were significantly lower than in those with elevated concentrations (P < 0.001). In ileal CD, ileal SES-CD correlated with histology (P < 0.001), but not with faecal calprotectin (P = 0.161) or lactoferrin (P = 0.448).In ileocolonic and colonic disease, endoscopic score SES-CD and histological findings correlated significantly with faecal calprotectin and lactoferrin. A normal faecal-marker concentration was a reliable surrogate marker for endoscopically and histologically inactive CD. Ileal endoscopic score and histological findings failed, however, to correlate with faecal markers.

Fifty-four infants with the malabsorption syndrome and cow's milk intolerence seen during 1962-1971 were investigated. All had diarrhoea and failed to thrive. Most had vomiting and about 20% had atopic eczema and recurrent respiratory infections. Laboratory investigations revealed malabsorption, raised serum IgA, and precipitins to cow's milk. Biopsies showed that the jejunal mucosa was damaged, and in about half the cases was flat. The patient did well on human milk but reacted clinically to cow's milk challenge, either in a few hours or gradually during 3-4 weeks. Some patients showed first a quick, but later a slow, reaction. Clinical symptoms of cow's milk intolerance disappeared at the age of about one year. At that time 81% had normal faecal fat, but only 29% had a normal proximal jejunal mucosa. Many of the patients developed intolerances to other food proteins, such as soya and wheat, if these were given during the sensitive period. Forty-two patients have been followed up for 2 years on a normal gluten-containing diet. Of these, 37 have a normal or nearly normal jejunal mucosa and 5 (12%) have subtotal villous atrophy indicative of coeliac disease. It is concluded that the malabsorption syndrome with cow's milk intolerance is a clear-cut clinical entity. However, the symptomatology, results of laboratory tests, and jejunal biopsy findings closely resemble those of other entities where damage to the intestinal mucosa causes a malabsorption snydrome. Follow-up studies showed that the disease is transient, but about 10% of the patients have coeliac disease, regarded in such cases as the primary disorder.

Early exposure to complex dietary proteins may increase the risk of beta-cell autoimmunity and type 1 diabetes in children with genetic susceptibility. We tested the hypothesis that supplementing breast milk with highly hydrolyzed milk formula would decrease the cumulative incidence of diabetes-associated autoantibodies in such children.In this double-blind, randomized trial, we assigned 230 infants with HLA-conferred susceptibility to type 1 diabetes and at least one family member with type 1 diabetes to receive either a casein hydrolysate formula or a conventional, cow's-milk-based formula (control) whenever breast milk was not available during the first 6 to 8 months of life. Autoantibodies to insulin, glutamic acid decarboxylase (GAD), the insulinoma-associated 2 molecule (IA-2), and zinc transporter 8 were analyzed with the use of radiobinding assays, and islet-cell antibodies were analyzed with the use of immunofluorescence, during a median observation period of 10 years (mean, 7.5). The children were monitored for incident type 1 diabetes until they were 10 years of age.The unadjusted hazard ratio for positivity for one or more autoantibodies in the casein hydrolysate group, as compared with the control group, was 0.54 (95% confidence interval [CI], 0.29 to 0.95), and the hazard ratio adjusted for an observed difference in the duration of exposure to the study formula was 0.51 (95% CI, 0.28 to 0.91). The unadjusted hazard ratio for positivity for two or more autoantibodies was 0.52 (95% CI, 0.21 to 1.17), and the adjusted hazard ratio was 0.47 (95% CI, 0.19 to 1.07). The rate of reported adverse events was similar in the two groups.Dietary intervention during infancy appears to have a long-lasting effect on markers of beta-cell autoimmunity--markers that may reflect an autoimmune process leading to type 1 diabetes. (ClinicalTrials.gov number, NCT00570102.).

Infants born by caesarean section or receiving antibiotics are at increased risk of developing metabolic, inflammatory and immunological diseases, potentially due to disruption of normal gut microbiota at a critical developmental time window. We investigated whether probiotic supplementation could ameliorate the effects of antibiotic use or caesarean birth on infant microbiota in a double blind, placebo-controlled randomized clinical trial. Mothers were given a multispecies probiotic, consisting of Bifidobacterium breve Bb99 (Bp99 2 × 108 cfu) Propionibacterium freundenreichii subsp. shermanii JS (2 × 109cfu), Lactobacillus rhamnosus Lc705 (5 × 109 cfu) and Lactobacillus rhamnosus GG (5 × 109 cfu) (N = 168 breastfed and 31 formula-fed), or placebo supplement (N = 201 breastfed and 22 formula-fed) during pregnancy, and the infants were given the same supplement. Faecal samples of the infants were collected at 3 months and analyzed using taxonomic, metagenomic and metaproteomic approaches. The probiotic supplement had a strong overall impact on the microbiota composition, but the effect depended on the infant’s diet. Only breastfed infants showed the expected increase in bifidobacteria and reduction in Proteobacteria and Clostridia. In the placebo group, both birth mode and antibiotic use were significantly associated with altered microbiota composition and function, particularly reduced Bifidobacterium abundance. In the probiotic group, the effects of antibiotics and birth mode were either completely eliminated or reduced. The results indicate that it is possible to correct undesired changes in microbiota composition and function caused by antibiotic treatments or caesarean birth by supplementing infants with a probiotic mixture together with at least partial breastfeeding. clinicaltrials.gov NCT00298337 . Registered March 2, 2006.

Cow's milk has been implicated as a possible trigger of the autoimmune response that destroys pancreatic beta cells in genetically susceptible hosts, thus causing diabetes mellitus. Studies in animals have suggested that bovine serum albumin (BSA) is the milk protein responsible, and an albumin peptide containing 17 amino acids (ABBOS) may be the reactive epitope. Antibodies to this peptide react with p69, a beta-cell surface protein that may represent the target antigen for milk-induced beta-cell—specific immunity.

BackgroundLarge, prospective population-based studies on clinical course, development of tolerance, and risk for other atopy in children with cow's milk allergy (CMA) are lacking.ObjectiveWe investigated the development of tolerance and the risk for asthma, rhinoconjunctivitis, atopic dermatitis, and sensitization in children with CMA followed to school age.MethodsWe followed 118 children with CMA until recovery and repeatedly measured their sensitization to cow's milk (CM). At age 8.6 years, 94 allergic subjects and 80 control subjects from the same cohort were studied for atopic diseases and sensitization. In addition, the parents of 12 allergic subjects and 26 control children returned a questionnaire on atopy, respectively.ResultsIgE-mediated CMA was detected in 86 (73%) children; at age 8.6 years, 13 (15%) had persistent CMA. All children with IgE-negative CMA were tolerant by age 5.0 years (P < .0001). Risk factors for persistent CMA at age 2.0 years were sensitization to CM at age 1.6 years (odds ratio, 6.3; 95% CI, 2.6-15.2), urticaria at diagnostic challenge (odds ratio, 3.3; 95% CI, 1.4-7.8), CM exposure at the maternity hospital (odds ratio, 3.2; 95% CI, 1.4-7.8), and early sensitization to egg (odds ratio, 2.8; 95% CI, 1.2-6.6). By age 8.6 years, children with IgE-positive CMA more frequently had asthma (31% vs 13%, P ≤ .01), rhinoconjunctivitis (66% vs 21%, P ≤ .001), atopic eczema (81% vs 26%, P ≤ .001), and sensitization to any allergen (88% vs 39%, P ≤ .001) than control subjects. CMA and family history of atopy were independent risk factors for atopic diseases, and CMA was also a risk factor for sensitization to inhalant allergens.ConclusionIgE-mediated CMA often persists to school age and is a risk factor for other atopy; non–IgE-mediated CMA, by contrast, is a benign infantile condition. Large, prospective population-based studies on clinical course, development of tolerance, and risk for other atopy in children with cow's milk allergy (CMA) are lacking. We investigated the development of tolerance and the risk for asthma, rhinoconjunctivitis, atopic dermatitis, and sensitization in children with CMA followed to school age. We followed 118 children with CMA until recovery and repeatedly measured their sensitization to cow's milk (CM). At age 8.6 years, 94 allergic subjects and 80 control subjects from the same cohort were studied for atopic diseases and sensitization. In addition, the parents of 12 allergic subjects and 26 control children returned a questionnaire on atopy, respectively. IgE-mediated CMA was detected in 86 (73%) children; at age 8.6 years, 13 (15%) had persistent CMA. All children with IgE-negative CMA were tolerant by age 5.0 years (P < .0001). Risk factors for persistent CMA at age 2.0 years were sensitization to CM at age 1.6 years (odds ratio, 6.3; 95% CI, 2.6-15.2), urticaria at diagnostic challenge (odds ratio, 3.3; 95% CI, 1.4-7.8), CM exposure at the maternity hospital (odds ratio, 3.2; 95% CI, 1.4-7.8), and early sensitization to egg (odds ratio, 2.8; 95% CI, 1.2-6.6). By age 8.6 years, children with IgE-positive CMA more frequently had asthma (31% vs 13%, P ≤ .01), rhinoconjunctivitis (66% vs 21%, P ≤ .001), atopic eczema (81% vs 26%, P ≤ .001), and sensitization to any allergen (88% vs 39%, P ≤ .001) than control subjects. CMA and family history of atopy were independent risk factors for atopic diseases, and CMA was also a risk factor for sensitization to inhalant allergens. IgE-mediated CMA often persists to school age and is a risk factor for other atopy; non–IgE-mediated CMA, by contrast, is a benign infantile condition.

Early feeding with cow's milk (CM) may increase the risk of cow's milk allergy (CMA).We sought to examine prospectively whether supplementary feeding of CM at the maternity hospital would increase the risk when compared with feeding with pasteurized human milk or hydrolyzed formula.We studied 6209 unselected healthy, full-term infants, of whom 5385 (87%) required supplementary milk while in the hospital. The infants were randomly assigned to receive CM formula (1789 infants), pasteurized human milk (1859 infants), or whey hydrolysate formula (1737 infants). The comparison group (824 infants) was composed of infants who were exclusively breast-fed. The infants were followed for 18 to 34 months for symptoms suggestive of CMA. The primary endpoint was a challenge-proven adverse reaction to CM after a successful CM elimination diet.The cumulative incidence of CMA in the infants fed CM was 2.4% compared with 1.7% in the pasteurized human milk group (odds ratio [OR], 0.70; 95% confidence interval [CI], 0. 44-1.12) and 1.5% in the whey hydrolysate group (OR, 0.61; 95% CI, 0. 38-1.00). In the comparison group, CMA developed in 2.1% of the infants. Among the infants who required supplementary feeding at hospital, both exposure to CM while in the hospital (OR, 1.54; 95% CI, 1.04-2.30; P =.03) and obvious parental atopy (OR, 2.32; 95% CI, 1.53-3.52; P <.001) increased the risk of CMA.Our data indicate that feeding of CM at maternity hospitals increases the risk of CMA when compared with feeding of other supplements, but exclusive breast-feeding does not eliminate the risk.

Fecal calprotectin and lactoferrin are promising noninvasive biomarkers for intestinal inflammation. In Crohn's disease (CD), during anti-TNF-alpha (TNF-alpha) treatment, the clinical significance of these markers has, however, been insufficiently explored.Among CD patients receiving anti-TNF-alpha therapy we assessed the role of fecal calprotectin and lactoferrin as surrogate markers for mucosal healing. Before and 3 months after the beginning of anti-TNF-alpha induction, 15 patients underwent ileocolonoscopy with scoring of the Crohn's Disease Index of Severity (CDEIS). Fecal samples for calprotectin and for lactoferrin measurements were collected and the Crohn's Disease Activity Index (CDAI) was calculated at the time of the endoscopies and 2 and 8 weeks after the first treatment.The median CDEIS fell from 13.0 to 4.8 (P = 0.002) and CDAI from 158 to 68 (P = 0.005). Accordingly, the median fecal calprotectin concentration fell from 1173 microg/g to 130 microg/g (P = 0.001) and fecal lactoferrin from 105.0 microg/g to 2.7 microg/g (P = 0.001). Of the 15 patients, 11 (73%) showed an endoscopic response to treatment and 5 of these achieved endoscopic remission (CDEIS < 3). In those 5 patients the fecal calprotectin concentration declined from 1891 mug/g (range 813-2434) to 27 microg/g (13-130) and lactoferrin from 92.4 microg/g (35.5-235.6) to 1.9 microg/g (0.0-2.1).Compared to pretreatment values, concentrations of fecal calprotectin and lactoferrin after the anti-TNF-alpha treatment were significantly lower. During anti-TNF-alpha therapy these fecal neutrophil-derived proteins may thus be useful surrogate markers for mucosal healing.

An unselected population of 8,954 children, age 8 to 15 years, was screened for hematuria. Four urine specimens from each were examined; microscopic hematuria was found in one or more specimens in 4.1%, and in two or more specimens in 1.1% of the children. The prevalence was not age or sex dependent. Those with two or more positive samples were re-examined twice during a half-year period: 33 had hematuria of 6 or more RBC/0.9 mm³, or more than 100,000 RBC/hour, on both occasions; renal biopsy performed on 28 of them revealed two cases of IgA-IgG nephropathy, one of focal segmental sclerosis, one of extracapillary glomerulonephritis, and one of possible hereditary nephritis. In 12 patients the biopsy was entirely normal; the rest showed equivocal changes. Co-existing proteinuria and the degree of hematuria correlated well with the severity of the morphologic alterations. Pathologic findings in microscopic hematuria seem to be less frequent than in hematuria in general; in most such patients, renal biopsy is probably not indicated. In some children the low-grade hematuria may merely represent the upper end of the physiologic variation.

Antigliadin antibodies in serum samples of 31 children with coeliac disease were measured by an enzyme-linked immunosorbent technique. In young patients (<2 years) tested before gluten withdrawal IgA antigliadin antibody levels were invariably above the levels of 36 controls. The titres fell rapidly when gluten was eliminated from the diet and rose on its reintroduction. The titres were not always greater than the control level in older untreated patients. IgA antigliadin antibodies seem to be a good marker of the immune reaction in the jejunum triggered by gluten. In 2 IgA-deficient patients gluten challenge caused an increase in IgM antigliadin antibodies, and at the same time the number of IgM-containing cells increased in the jejunal mucosa. Rising IgG antigliadin antibody levels after gluten elimination were seen in 6 patients, 5 of whom had very low complement C3 levels before gluten elimination.

Live probiotic bacteria and dietary prebiotic oligosaccharides (together termed synbiotics) increasingly are being used in infancy, but evidence of long-term safety is lacking. In a randomized, placebo-controlled, double-blind trial, we studied the safety and long-term effects of feeding synbiotics to newborn infants.Between November 2000 and March 2003, pregnant mothers carrying infants at high risk for allergy were randomly assigned to receive a mixture of 4 probiotic species (Lactobacillus rhamnosus GG and LC705, Bifidobacterium breve Bb99, and Propionibacterium freudenreichii ssp shermanii) or a placebo for 4 weeks before delivery. Their infants received the same probiotics with 0.8 g of galactooligosaccharides, or a placebo, daily for 6 months after birth. Safety data were obtained from clinical examinations and interviews at follow-up visits at ages 3, 6, and 24 months and from questionnaires at ages 3, 6, 12, and 24 months. Growth data were collected at each time point.Of the 1018 eligible infants, 925 completed the 2-year follow-up assessment. Infants in both groups grew normally. We observed no difference in neonatal morbidity, feeding-related behaviors (such as infantile colic), or serious adverse events between the study groups. During the 6-month intervention, antibiotics were prescribed less often in the synbiotic group than in the placebo group (23% vs 28%). Throughout the follow-up period, respiratory infections occurred less frequently in the synbiotic group (geometric mean: 3.7 vs 4.2 infections).Feeding synbiotics to newborn infants was safe and seemed to increase resistance to respiratory infections during the first 2 years of life.

We studied the balance between ileal T-effector cells versus T-regulatory cells in active and inactive Crohn's disease (CD).We compared effector and regulatory T-cell-related markers such as interleukin (IL)-17, interferon (IFN)-gamma, IL-4, and Foxp3 transforming growth factor (TGF)-beta CTLA-4 and markers for innate immune activation such as IL-6, IL-10, IL-18, IL-23, tumor necrosis factor (TNF)-alpha, and IL-12p70, studied with immunohistochemistry and RT-PCR in ileal biopsies from patients with active or inactive CD and from control subjects. IL-17 in fecal samples was detected by ELISA. The effect of IL-17 on IL-8 and TNF-alpha mRNA expression in epithelial cell line Caco-2 was studied.The numbers of IL-4-, IL-17-, and IL-23(p19)-positive cells in the lamina propria were higher in patients with CD, both active and inactive, than in the controls. mRNA expression of IL-17A, IL-6, and Foxp3 was increased in the biopsies both from patients with active disease and those in remission, whereas mRNA expression of IL-23 was increased only in active disease. Fecal IL-17 concentration was increased in patients with active disease. IL-17 enhanced the IL-8 and TNF-alpha response of the epithelial cell line to lipopolysaccharide (LPS) in vitro.Our findings suggest that activation of the IL-23/IL-17 axis is fundamentally connected to the etiology of CD and may represent the basis for the relapsing nature of the disease by increasing the sensitivity of epithelium to microbial LPS.

Coeliac disease may remain undiagnosed because of the non-specific nature of the presenting symptoms. Several antibody tests are claimed as markers for this condition but a direct comparison of the available tests has not been reported. The probands and healthy first-degree relatives of 42 families with coeliac disease were studied. Histological examination of biopsy specimens revealed jejunal mucosal villous atrophy compatible with coeliac disease in 13 of 122 relatives. Reticulin-antibody-positive relatives with or without jejunal mucosal atrophy were genetically similar to the probands of the families (DR3 gene frequencies 55.3%-60.0%). Gliadin-antibody-positive relatives with normal mucosa were genetically different from the probands (DR3 gene frequency 16.7% versus 55.3%). IgA reticulin and endomysium antibodies detected 92.3% of subjects with silent coeliac disease. The only case that was missed had selective IgA deficiency and was positive for IgG-class reticulin antibodies. By contrast, gliadin antibodies detected only half of the cases. Follow-up of the 7 reticulin-antibody-positive relatives with normal mucosa revealed 2 further cases of coeliac disease and 1 of dermatitis herpetiformis during the next three years. Our family study shows that healthy reticulin-antibody-positive first-degree relatives of coeliac disease patients, irrespective of the state of the jejunal mucosa, are genetically similar to known coeliac disease patients. Reticulin-antibody positivity is an indicator of both silent and latent coeliac disease.

The mechanism of the developmental downregulation of the lactase-phlorizin hydrolase (LPH) gene underlying adult-type hypolactasia is unknown. We have determined the functional significance of the recently identified two single nucleotide polymorphisms (SNPs), C/T(-13910) and G/A(-22018), associated with adult-type hypolactasia by studying LPH mRNA levels in intestinal biopsy samples with different genotypes.Intestinal biopsy samples were taken from 52 patients with abdominal complaints. Hypolactasia was diagnosed by determining lactase and sucrase activities and calculating their ratio (L/S ratio). The functional effect of the C/T(-13910) and G/A(-22018) genotype on expression of LPH mRNA was demonstrated in patients heterozygous for the C/T(-13910) and G/A(-22018) polymorphism and an informative expressed SNP located in the coding region of the LPH mRNA. Reverse transcription-polymerase chain reaction followed by solid phase minisequencing was used for accessing the relative expression levels of the LPH alleles using informative SNPs located in exons 1, 2, 6, 10, 13, or 17 as markers.Statistically significant differences between the three different genotypes CC(-13910) GG(-22018), CT(-13910) GA(-22018), and TT(-13910) AA(-22018) and their respective L/S ratios were observed. Relative quantitation of the expressed LPH alleles showed that the persistent allele represented 92 (6)% (mean (SEM), range 78-99%; n=14) of the expressed LPH mRNA. The patient with the homozygous persistent TT(-13910) AA(-22018), as well as hypolactasic patients with CC(-13910) GG(-22018), showed equal expression of both alleles (47 (1)%; n=7).Expression of LPH mRNA in the intestinal mucosa in individuals with T(-13910) A(-22018) alleles is several times higher than that found in individuals with C(-13910), G(-22018) alleles. These findings suggest that the two SNPs, C/T(-13910) and G/A(-22018), associated with adult-type hypolactasia, are associated with the transcriptional regulation of the LPH gene. The presence of the T(-13910) A(-22018) allele also shows significant elevation of the L/S ratio.

A jejunal biopsy specimen from an asympto- matic 35 year old man was studied because of a low serum titre of reticulin antibody and the finding of coeliac disease in his son.In this specimen villous structure was quite normal as was the total number of intraepithelial lymphocytes, but the number of y/b T cell receptor bearing lymphocytes was 10 times higher than the mean in control subjects.Two years later a further biopsy specimen was obtained because of clinical symptoms and an increased titre of reticulin antibody.This specimen showed villous atrophy with crypt hyperplasia and increased infiltration of intraepithelial lymphocytes compatible with coeliac disease.A con- trol biopsy specimen taken during gluten free diet showed normalisation of the villous archi- tecture.Latent coeliac disease may be charac- terised by an increase in y/6 positive cells similar to that seen in established coeliac disease.

Probiotics are widely studied both in the treatment and prevention of allergic diseases, but their mode of action is poorly known.Our aim was to examine the effect of probiotic bacteria on in vivo cytokine, antibody, and inflammatory responses in allergy-prone infants.In a randomized double-blind study, probiotic bacteria or placebo were given for 1 month before delivery to mothers and for 6 months to infants with a family history of allergy. Plasma samples were analysed for C-reactive protein (CRP), total IgA and IgE, food-specific IgA, IgG, and IgE, IL-2, IL-4, IL-6, IL-10, TNF-alpha, and IFN-gamma. We analysed the associations of immunological and inflammatory parameters at age 6 months with probiotic treatment and allergic phenotype at 2 years.Infants receiving probiotic bacteria had higher plasma levels of CRP (P=0.008), total IgA (P=0.016), total IgE (P=0.047), and IL-10 (P=0.002) than infants in the placebo group. Increased plasma CRP level at age 6 months was associated with a decreased risk of eczema [odds ratio (OR) 0.41 [95% confidence interval (CI) 0.17-0.99], P=0.046], and with a decreased risk of allergic disease [OR 0.38 (95% CI 0.16-0.87), P=0.023] at age 2 years, when adjusted with probiotic use.The association of CRP with a decreased risk of eczema at 2 years of age in allergy-prone children supports the view that chronic, low-grade inflammation protects from eczema. Probiotic-induced low-grade inflammation was characterized by elevation of IgE, IgA, and IL-10, the changes typically observed in helminth infection-associated induction of regulatory mechanisms. The findings emphasize the role of chronic microbial exposure as an immune modulator protecting from allergy.

Involvement of gut immune system has been implicated in the pathogenesis of type 1 diabetes. However, few studies have been performed on the gut mucosa from patients with type 1 diabetes. Thus, we characterized the stage of immune activation in jejunal biopsy samples from 31 children with type 1 diabetes by immunohistochemistry, in situ hybridization, and RT-PCR. We found enhanced expressions of HLA-DR, HLA-DP, and intercellular adhesion molecule-1 by immunohistochemistry even on structurally normal intestine of patients with type 1 diabetes and no signs of celiac disease. In addition, the densities of IL-1 alpha- and IL-4-positive cells detected by immunohistochemistry and IL-4 mRNA-expressing cells evaluated by in situ hybridization were increased in the lamina propria in patients with type 1 diabetes and normal mucosa. Instead, the densities of IL-2, gamma-interferon (IFN-gamma), and tumor necrosis factor alpha-positive cells, the density of IFN-gamma mRNA positive cells, and the amounts of IFN-gamma mRNA detected by RT-PCR correlated with the degree of celiac disease in patients with type 1 diabetes. Our study supports the hypothesis that a link exists between the gut immune system and type 1 diabetes.

Probiotic bacteria are proposed to alleviate intestinal inflammation in infants with atopic eczema/dermatitis syndrome (AEDS) and food allergy. In such infants we investigated effects of probiotic bacteria on faecal IgA, and on the intestinal inflammation markers tumour necrosis factor-alpha (TNF-alpha), alpha1-antitrypsin (AT), and eosinophil cationic protein (ECP). A total of 230 infants with AEDS and suspected cow's milk allergy (CMA) received in a randomized double-blinded manner, concomitant with elimination diet, Lactobacillus GG (LGG), a mixture of four probiotic strains (MIX), or placebo for 4 wk. Four weeks after treatment, CMA was diagnosed with a double-blind placebo-controlled milk challenge. Faecal samples of 102 infants, randomly chosen for analysis, were collected before treatment, after 4-wk treatment, and on the first day of milk challenge. After treatment, IgA levels tended to be higher in probiotic groups than in the placebo group (LGG vs. placebo, p=0.064; MIX vs. placebo, p=0.064), and AT decreased in the LGG group, but not in other treatment groups. After challenge in IgE-associated CMA infants, faecal IgA was higher for LGG than for placebo (p=0.014), and TNF-alpha was lower for LGG than for placebo, but non-significantly (p=0.111). In conclusion, 4-wk treatment with LGG may alleviate intestinal inflammation in infants with AEDS and CMA.

The immunomodulating mechanisms of Lactobacillus GG (LGG) and other probiotics are poorly understood.We studied in vivo the immunologic effects of probiotics in infants with atopic eczema-dermatitis syndrome (AEDS) and cow's milk allergy (CMA).Two hundred thirty infants with AEDS and suspected CMA received, concomitant with elimination diet, either LGG, a mixture of 4 probiotic strains (MIX), or placebo for 4 weeks. All available paired pretreatment and posttreatment plasma samples (n = 132) were analyzed for concentrations of IL-2, IL-4, IL-6, IL-10, TNF-alpha, IFN-gamma, soluble intercellular adhesion molecule 1, soluble E-selectin, TGF-beta1, TGF-beta2, and C-reactive protein.In infants with IgE-associated AEDS, treatment with LGG induced higher C-reactive protein levels than in the placebo group (geometric mean, 0.83 microg/mL [95% CI, 0.56-0.81] vs 0.42 microg/mL [95% CI, 0.27-0.65]; P = .021). Concomitantly, IL-6 levels increased after treatment with LGG ( P = .023) but not with MIX or placebo. Soluble E-selectin levels were higher after probiotic than after placebo treatment in infants with IgE-mediated CMA (LGG geometric mean, 86.7 ng/mL [95% CI, 75.2-100]; MIX geometric mean, 91.6 ng/mL [95% CI, 74.8-111.9]; and placebo geometric mean, 64.9 ng/mL [95% CI, 53-79.3]; analysis of covariance, P = .035; LGG vs placebo, P = .023; MIX vs placebo, P = .020). Use of MIX induced an increase in plasma IL-10 levels ( P = .016).Probiotics induced systemically detectable low-grade inflammation, which might explain the clinical effects of probiotics in AEDS and CMA.

A single-nucleotide variant, C/T-13910, located 14 kb upstream of the lactase gene (LCT), has been shown to be completely correlated with lactase persistence (LP) in northern Europeans. Here, we analyzed the background of the alleles carrying the critical variant in 1,611 DNA samples from 37 populations. Our data show that the T-13910 variant is found on two different, highly divergent haplotype backgrounds in the global populations. The first is the most common LP haplotype (LP H98) present in all populations analyzed, whereas the others (LP H8–H12), which originate from the same ancestral allelic haplotype, are found in geographically restricted populations living west of the Urals and north of the Caucasus. The global distribution pattern of LP T-13910 H98 supports the Caucasian origin of this allele. Age estimates based on different mathematical models show that the common LP T-13910 H98 allele (∼5,000–12,000 years old) is relatively older than the other geographically restricted LP alleles (∼1,400–3,000 years old). Our data about global allelic haplotypes of the lactose-tolerance variant imply that the T-13910 allele has been independently introduced more than once and that there is a still-ongoing process of convergent evolution of the LP alleles in humans. A single-nucleotide variant, C/T-13910, located 14 kb upstream of the lactase gene (LCT), has been shown to be completely correlated with lactase persistence (LP) in northern Europeans. Here, we analyzed the background of the alleles carrying the critical variant in 1,611 DNA samples from 37 populations. Our data show that the T-13910 variant is found on two different, highly divergent haplotype backgrounds in the global populations. The first is the most common LP haplotype (LP H98) present in all populations analyzed, whereas the others (LP H8–H12), which originate from the same ancestral allelic haplotype, are found in geographically restricted populations living west of the Urals and north of the Caucasus. The global distribution pattern of LP T-13910 H98 supports the Caucasian origin of this allele. Age estimates based on different mathematical models show that the common LP T-13910 H98 allele (∼5,000–12,000 years old) is relatively older than the other geographically restricted LP alleles (∼1,400–3,000 years old). Our data about global allelic haplotypes of the lactose-tolerance variant imply that the T-13910 allele has been independently introduced more than once and that there is a still-ongoing process of convergent evolution of the LP alleles in humans. The expression of the lactase enzyme (MIM 603202) in intestinal cells dramatically declines after weaning in mammals, when lactose is no longer an essential part of their diet.1Sahi T Isokoski M Jussila J Launiala K Pyorala K Recessive inheritance of adult-type lactose malabsorption.Lancet. 1973; 2: 823-826Abstract PubMed Scopus (127) Google Scholar In humans, this normal mammalian condition known as “lactase nonpersistence” (LNP, also known as “adult-type hypolactasia” or “lactose intolerance” [MIM 223100]) affects most of mankind and restricts the consumption of fresh milk among adults. However, among northern Europeans and a few other ethnic populations, intestinal lactase activity persists throughout life in a substantial proportion (up to 80%–90%) of adults, a condition known as lactase persistence (LP, or lactose tolerance [MIM 223100]). The LP/LNP phenotype is genetically determined, with LP being dominant over LNP.2Sahi T Genetics and epidemiology of adult-type hypolactasia.Scand J Gastroenterol Suppl. 1994; 202: 7-20Crossref PubMed Scopus (269) Google Scholar We previously identified a single-nucleotide variant, C/T-13910, completely correlating with the phenotype in Finns and in a cross-sectional sample of >600 individuals from five populations.3Enattah NS Sahi T Savilahti E Terwilliger JD Peltonen L Jarvelä I Identification of a variant associated with adult-type hypolactasia.Nat Genet. 2002; 30: 233-237Crossref PubMed Scopus (828) Google Scholar, 4Kuokkanen M Enattah NS Oksanen A Savilahti E Orpana A Jarvela I Transcriptional regulation of the lactase-phlorizin hydrolase gene by polymorphisms associated with adult-type hypolactasia.Gut. 2003; 52: 647-652Crossref PubMed Scopus (164) Google Scholar, 5Rasinpera H Savilahti E Enattah NS Kuokkanen M Totterman N Lindhal H Jarvela I Kolho K-L A genetic test which can be used to diagnose adult-type hypolactasia in children.Gut. 2004; 53: 1571-1576Crossref PubMed Scopus (162) Google Scholar The T-13910 variant, which correlates with LP, is located 14 kb upstream of the LCT gene and has been shown to be the derived variant, compared with the C-13910 variant that represents the ancestral form of the human genome. Another variant, G/A-22018, farther upstream of LCT, was also strongly, although not completely, associated with the LP/LNP phenotype,3Enattah NS Sahi T Savilahti E Terwilliger JD Peltonen L Jarvelä I Identification of a variant associated with adult-type hypolactasia.Nat Genet. 2002; 30: 233-237Crossref PubMed Scopus (828) Google Scholar, 5Rasinpera H Savilahti E Enattah NS Kuokkanen M Totterman N Lindhal H Jarvela I Kolho K-L A genetic test which can be used to diagnose adult-type hypolactasia in children.Gut. 2004; 53: 1571-1576Crossref PubMed Scopus (162) Google Scholar most likely because of the substantial linkage disequilibrium (LD) in this genome region.3Enattah NS Sahi T Savilahti E Terwilliger JD Peltonen L Jarvelä I Identification of a variant associated with adult-type hypolactasia.Nat Genet. 2002; 30: 233-237Crossref PubMed Scopus (828) Google Scholar, 6Harvey CB Pratt WS Islam I Whitehouse DB Swallow DM DNA polymorphisms in the lactase gene: linkage disequilibrium across the 70-kb region.Eur J Hum Genet. 1995; 3: 27-41Crossref PubMed Scopus (36) Google Scholar, 7Harvey CB Hollox EJ Poulter M Wang Y Rossi M Auricchio S Iqbal TH Cooper BT Barton R Sarner M et al.Lactase haplotype frequencies in Caucasians: association with the lactase persistence/non-persistence polymorphism.Ann Hum Genet. 1998; 62: 215-223Crossref PubMed Google Scholar, 8Poulter M Hollox E Harvey CB Mulcare C Peuhkuri K Kajander K Sarner M Korpela R Swallow DM The causal element for the lactase persistence/non-persistence polymorphism is located in a 1 Mb region of linkage disequilibrium in Europeans.Ann Hum Genet. 2003; 67: 298-311Crossref PubMed Scopus (109) Google Scholar, 9Bersaglieri T Sabeti PC Patterson N Vanderploeg T Schaffner SF Drake JA Rhodes M Reich DE Hirschhorn JN Genetic signatures of strong recent positive selection at the lactase gene.Am J Hum Genet. 2004; 74: 1111-1120Abstract Full Text Full Text PDF PubMed Scopus (740) Google Scholar Functional evidence for the C/T-13910 variant in the regulation of lactase activity has since emerged, lending additional support for this nucleotide change as the true causative variant of regulation of transcription of the lactase gene in intestinal cells.4Kuokkanen M Enattah NS Oksanen A Savilahti E Orpana A Jarvela I Transcriptional regulation of the lactase-phlorizin hydrolase gene by polymorphisms associated with adult-type hypolactasia.Gut. 2003; 52: 647-652Crossref PubMed Scopus (164) Google Scholar, 10Olds LC Sibley E Lactase persistence DNA variant enhances lactase promoter activity in vitro: functional role as a cis regulatory element.Hum Mol Genet. 2003; 12: 2333-2340Crossref PubMed Scopus (179) Google Scholar, 11Troelsen JT Olsen J Moller J Sjostrom H An upstream polymorphism associated with lactase persistence has increased enhancer activity.Gastroenterology. 2003; 125: 1686-1694Abstract Full Text Full Text PDF PubMed Scopus (152) Google Scholar Adult individuals with the LP T-13910 allele show significantly higher steady-state transcript levels of LCT in their intestinal mucosa when compared with individuals with the nonpersistence C-13910 allele, which implies a transcriptional regulation of LCT.4Kuokkanen M Enattah NS Oksanen A Savilahti E Orpana A Jarvela I Transcriptional regulation of the lactase-phlorizin hydrolase gene by polymorphisms associated with adult-type hypolactasia.Gut. 2003; 52: 647-652Crossref PubMed Scopus (164) Google Scholar This is in agreement with in vitro studies demonstrating a distinct increase in the LCT promoter activity in cells transfected with the T-13910 variant.10Olds LC Sibley E Lactase persistence DNA variant enhances lactase promoter activity in vitro: functional role as a cis regulatory element.Hum Mol Genet. 2003; 12: 2333-2340Crossref PubMed Scopus (179) Google Scholar, 11Troelsen JT Olsen J Moller J Sjostrom H An upstream polymorphism associated with lactase persistence has increased enhancer activity.Gastroenterology. 2003; 125: 1686-1694Abstract Full Text Full Text PDF PubMed Scopus (152) Google Scholar, 12Lewinsky RH Jensen TG Moller J Stensballe A Olsen J Troelsen JT T-13910 DNA variant associated with lactase persistence interacts with Oct-1 and stimulates lactase promoter activity in vitro.Hum Mol Genet. 2005; 14: 3945-3953Crossref PubMed Scopus (139) Google Scholar Haplotype analysis in the Finnish families demonstrated that all LP alleles among Finns originated from one common ancestor identical by descent.3Enattah NS Sahi T Savilahti E Terwilliger JD Peltonen L Jarvelä I Identification of a variant associated with adult-type hypolactasia.Nat Genet. 2002; 30: 233-237Crossref PubMed Scopus (828) Google Scholar Other studies of additional European populations have also suggested the existence of one major allelic haplotype, named “haplotype A,” correlating with LP.7Harvey CB Hollox EJ Poulter M Wang Y Rossi M Auricchio S Iqbal TH Cooper BT Barton R Sarner M et al.Lactase haplotype frequencies in Caucasians: association with the lactase persistence/non-persistence polymorphism.Ann Hum Genet. 1998; 62: 215-223Crossref PubMed Google Scholar, 13Hollox EJ Poulter M Zvarik M Ferak V Krause A Jenkins T Saha N Kozlov AI Swallow DM Lactase haplotype diversity in the Old World.Am J Hum Genet. 2001; 68: 160-172Abstract Full Text Full Text PDF PubMed Scopus (139) Google Scholar These data indicate a single global origin for the LP T-13910 allele. In this study, we monitored the global frequencies of the LP T-13910 allele and allelic haplotype signatures of the ∼30-kb LCT locus in diverse global populations, to study the allelic background of LP in humans. We genotyped eight SNPs and one indel polymorphism (GenBank accession number DQ109677) covering ∼30 kb of the LCT region and flanking the two LCT variants, C/T-13910 and G/A-22018, associated with LP/LNP (coverage rate of one SNP per 3.3 kb) in 1,611 samples from 37 global populations (fig. 1 and table 1). Except for the two SNPs C/T-13910 and G/A-22018, the genotyped SNPs represent common variants in all populations, with minor-allele frequencies >7% (table 2). Although this approach might not identify some rare allelic variants, especially among the LNP alleles, the most robust pattern of diversity among LP alleles—the target of our interest—will be identified in the global samples.Table 1SNPs Analyzed in the DNA Samples from 37 PopulationsSNPTypedbSNP Accession NumberChromosomal PositionDistance from LCT (kb)1IndelDQ109677aThe GenBank accession number is given.136424826+3.642C/Trs3754686136437008−8.543G/Crs3769005136437098−8.634C/T-13910rs4988235136442378−13.915C/Trs4954493136443707−15.2396G/Crs3099181136448545−20.0777G/A-22018rs182549136450486−22.0188C/Trs4988183136455779−27.3129A/Crs3087343136456274−27.807Note.—The 3,954-bp indel polymorphism is located within intron 1 of the LCT gene. Accurate chromosomal positions and locations from LCT are given. The SNPs are also shown in figure 1.a The GenBank accession number is given. Open table in a new tab Table 2SNP Frequencies Analyzed in 37 Population SamplesDerived Allele 2 Frequency (SD)Region or PopulationNSNP 1 (Indel; in=1, del=2)SNP 2 (C/T; T=1, G=2)SNP 3 (G/C; G=1, C=2)SNP 4 (C/T-13910; C=1, T=2)SNP 5 (C/T; T=1, C=2)SNP 6 (G/C; G=1, C=2)SNP 7 (G/A -22018; G=1, A=2)SNP 8 (A/C; C=1, A=2)SNP 9 (G/A; G=1, A=2)South Korea23.07 (.04).28 (.06).28 (.06).00 (.00).28 (.06).28 (.06).00 (.00).52 (.07).89 (.05)Han Chinese100.38 (.03).36 (.03).36 (.03).00 (.00).36 (.03).36 (.03).00 (.00).64 (.03).77 (.03)Ob-Ugric speakers20.45 (.04).43 (.05).43 (.05).03 (.02).43 (.05).42 (.02).03 (.02).43 (.05).81 (.04)Komi10.40 (.09).50 (.11).50 (.11).15 (.07).50 (.11).50 (.11).15 (.07).65 (.10).85 (.08)Udmurts30.53 (.06).48 (.07).48 (.07).33 (.06).50 (.06).50 (.06).37 (.07).58 (.06).85 (.04)Mokshas30.27 (.06).30 (.06).30 (.06).28 (.06).27 (.06).27 (.06).27 (.06).37 (.07).57 (.06)Erzas30.48 (.06).42 (.06).42 (.06).27 (.06).42 (.06).38 (.05).22 (.05).40 (.06).68 (.06)Saami30.53 (.06).51 (.07).51 (.07).17 (.04).51 (.07).51 (.07).13 (.04).60 (.06).85 (.05)Finns, eastern77.69 (.04).69 (.04).69 (.03).55 (.04).68 (.03).66 (.04).55 (.04).70 (.04).88 (.03)Finns, western154.73 (.02).71 (.03).71 (.03).62 (.02).72 (.02).71 (.03).62 (.02).73 (.02).88 (.02)Daghestan Druss17.23 (.07).23 (.07).18 (.07).12 (.06).21 (.07).21 (.07).12 (.06).26 (.07).62 (.09)Daghestan Nog20.40 (.08).40 (.08).37 (.08).07 (.04).40 (.08).40 (.08).07 (.04).40 (.08).60 (.08)Daghestan mixed23.35 (.07).35 (.07).35 (.07).13 (.05).35 (.07).35 (.07).13 (.05).37 (.07).67 (.07)Pakistan Balti23.24 (.06).17 (.06).26 (.06).00 (.00).17 (.06).17 (.06).00 (.00).26 (.07).44 (.07)Pakistan Burusho30.33 (.07).33 (.07).33 (.07).02 (.01).33 (.07).23 (.06).05 (.03).28 (.06).77 (.06)Pakistan Kashmiri20.37 (.08).42 (.08).42 (.08).12 (.05).42 (.08).37 (.08).15 (.06).42 (.08).78 (.07)Pakistan Kalash30.25 (.06).27 (.06).25 (.05).00 (.00).25 (.06).22 (.05).03 (.02).38 (.08).62 (.06)Pakistan Pathan28.45 (.07).41 (.07).43 (.07).30 (.06).39 (.07).41 (.07).32 (.06).48 (.07).71 (.06)Pakistan Hazara14.36 (.09).32 (.09).32 (.09).04 (.04).32 (.09).29 (.09).11 (.06).46 (.10).64 (.09)Pakistan Baluch19.47 (.08).47 (.08).47 (.08).34 (.08).47 (.08).47 (.08).39 (.08).50 (.08).79 (.06)Pakistan Sindi28.50 (.07).52 (.07).50 (.07).41 (.07).50 (.07).50 (.07).43 (.07).52 (.07).75 (.06)Pakistan Brahui30.43 (.07).42 (.07).43 (.06).27 (.06).43 (.06).40 (.06).28 (.06).43 (.07).82 (.05)Pakistan Makrani Baluch29.35 (.06).35 (.06).35 (.06).17 (.05).35 (.06).33 (.06).18 (.05).48 (.07).77 (.06)Pakistan Mohannes29.38 (.06).38 (.06).36 (.07).28 (.06).36 (.07).36 (.06).28 (.06).36 (.06).67 (.07)Pakistan Parsi29.21 (.05).19 (.05).19 (.05).14 (.05).22 (.06).17 (.05).03 (.02).33 (.06).55 (.07)Iranians21.26 (.07).21 (.06).21 (.06).10 (.05).24 (.07).21 (.06).07 (.04).26 (.07).57 (.08)Iran Qashqai10.10 (.07).10 (.07).10 (.07).05 (.05).10 (.07).10 (.07).05 (.05).10 (.07).40 (.11)Arabs50.17 (.04).25 (.04).18 (.04).10 (.03).16 (.04).15 (.04).10 (.03).19 (.04).52 (.05)Southern Italy100.22 (.03).26 (.03).25 (.03).05 (.02).26 (.03).26 (.03).06 (.02).29 (.04).60 (.03)French17.44 (.08).44 (.08).44 (.08).34 (.07).44 (.08).44 (.08).37 (.08).50 (.08).62 (.09)Basques85.71 (.03).70 (.03).73 (.03).66 (.04).72 (.04).69 (.03).64 (.04).70 (.03).86 (.03)Utah92.83 (.03).83 (.03).83 (.03).74 (.03).83 (.03).82 (.03).76 (.03).83 (.03).90 (.02)Somalia79.18 (.03).19 (.03).22 (.03).03 (.01).18 (.03).17 (.03).01 (.01).28 (.04).68 (.03)Fulani Sudanese44.80 (.05).57 (.06).57 (.05).48 (.06).56 (.06).56 (.05).55 (.06).82 (.05).94 (.02)Morocco90.35 (.03).33 (.04).33 (.03).18 (.03).35 (.03).33 (.03).16 (.03).41 (.04).64 (.03)Saharawi57.36 (.05).36 (.04).37 (.04).26 (.04).36 (.05).36 (.04).29 (.04).36 (.04).72 (.04)African Americans50.25 (.04).18 (.04).21 (.04).09 (.03).20 (.04).18 (.04).09 (.03).62 (.05).80 (.04)Note.—Alleles coded as 1 in every SNP site were the ancestral alleles, on the basis of the sequence of the primate samples, that cosegregated with the LNP phenotype, and alleles coded as 2 in every SNP site were the derived alleles that cosegregated with the LP phenotype. Open table in a new tab Note.— The 3,954-bp indel polymorphism is located within intron 1 of the LCT gene. Accurate chromosomal positions and locations from LCT are given. The SNPs are also shown in figure 1. Note.— Alleles coded as 1 in every SNP site were the ancestral alleles, on the basis of the sequence of the primate samples, that cosegregated with the LNP phenotype, and alleles coded as 2 in every SNP site were the derived alleles that cosegregated with the LP phenotype. The frequency of the LP T-13910 allele in various populations was systematically correlated with the reported prevalence of LP determined elsewhere by disaccharidase activities in intestinal biopsy samples and/or lactose-tolerance tests in these populations (fig. 2 and table 3).1Sahi T Isokoski M Jussila J Launiala K Pyorala K Recessive inheritance of adult-type lactose malabsorption.Lancet. 1973; 2: 823-826Abstract PubMed Scopus (127) Google Scholar, 2Sahi T Genetics and epidemiology of adult-type hypolactasia.Scand J Gastroenterol Suppl. 1994; 202: 7-20Crossref PubMed Scopus (269) Google Scholar, 14Simoons FJ The geographic hypothesis and lactose malabsorption: a weighing of the evidence.Am J Dig Dis. 1978; 23: 963-980Crossref PubMed Scopus (223) Google Scholar, 15Swallow DM Genetics of lactase persistence and lactose intolerance.Annu Rev Genet. 2003; 37: 197-219Crossref PubMed Scopus (386) Google Scholar, 16Holden C Mace R Phylogenetic analysis of the evolution of lactose digestion in adults.Hum Biol. 1997; 69: 605-628PubMed Google Scholar, 17Flatz G Rotthauwe HW The human lactase polymorphism: physiology and genetics of lactose absorption and malabsorption.Prog Med Genet. 1977; 2: 205-249PubMed Google Scholar Among the 37 populations studied (fig. 3), we identified 21 populations for which the prevalence of the LP trait was known and could establish a strong correlation (coefficient of correlation r=0.973, P<.0001) with the frequency of the T-13910 allele (fig. 2). The allele frequencies of the analyzed markers are shown in table 2, and the complete list of all observed haplotypes constructed using all nine markers with the Arlequin program18Schneider S Roessli D Excoffier L Arlequin version 2.000: a software for population genetic data analysis. Genetics and Biometry Laboratory, University of Geneva, Geneva2000Google Scholar are provided in table 4. We restricted further analysis to those haplotypes with population frequency >4% in at least one of the populations, as inferred by the Arlequin program, to avoid misleading conclusions based on rare haplotypes, which could represent artifacts of the algorithm used for the construction of the haplotypes (table 5). We identified 9 different haplotypes (H8, H9, H11, H12, H48, H49, H95, H97, and H98) with alleles carrying the T-13910 LP variant and 14 haplotypes (H1, H2, H4, H27, H34, H46, H51, H52, H54, H55, H81, H82, H84, and H87) with alleles carrying the C-13910 LNP variant (table 5). Comparison of the resulting haplotypes with the haplotypes estimated by the maximum-likelihood algorithm implemented in the PHASE program v2.1 did not reveal discrepancies (data not shown).Table 3Population Frequencies of LP Alleles C/T-1391No. with GenotypeAllele Frequency (%)DesignationRegion or PopulationThree-Letter CodeNCCCTTTCTPrevalence of LP (% [SD])1South KoreaSKo23230010000 (.00)2Han ChineseHan1001000010000 (.00)3Ob-Ugric speakersObU62584096.83.26 (3.02)4KomiKom10730851530 (14.50)5UdmurtsUdm301216266.633.460 (8.90)6MokshasMok301317071.628.456.6 (9.01)7ErzasErz301710373.326.743.3 (9.05)8SaamiSaa302010083.316.733.3 (8.60)9Finns, easternFiE7718352446.153.976.6 (4.75)10Finns, westernFiW15425686138.361.783.7 (2.98)11Daghestan DrussDaD17134088.211.823.5 (10.30)12Daghestan NogDaN20155087.512.525 (9.70)13Daghestan mixedDaM23193189.111.917.4 (7.90)14BaltiBal23230010000 (.00)15BurushoBur30291098.31.73.3 (3.26)16KashmiriKas20155087.512.525 (9.68)17KalashKal30300010000 (.00)18PathanPat281215169.630.457.1 (9.35)19HazaraHaz14131096.43.67.1 (6.86)20BaluchBlu19106368.431.647.4 (11.46)21SindiSin281013558.941.164.3 (9.11)22BrahuiBra301710373.326.743.3 (9.05)23Makrani BaluchMaB291910082.817.234.5 (8.83)24MohannesMoh291610372.427.644.8 (9.23)25ParsiPar29218086.213.827.6 (8.30)26IraniansIra21174090.59.519 (8.56)27QashqaiQas1091095510 (9.49)28ArabsAra51428190.29.817.6 (5.33)29Southern ItalySIt1008911094.55.511 (3.13)30FrenchFra1769161.738.358.8 (11.94)31BasquesBas857443434.165.991.7 (2.99)32UtahUta927335225.574.592.4 (2.76)33SomaliaSom79745096.83.26.3 (2.73)34Fulani SudaneseFul44132011524870.4 (6.88)35SaharawiSah572926273.726.349.1 (6.62)36MoroccoMor906225382.717.331.1 (4.88)37African AmericansAam50443391912 (4.60) Open table in a new tab Figure 3Population frequencies for the T-13910 allele associated with LP in worldwide populations. For each population, the pie chart denotes the frequency of the T-13910 allele (green shading). Populations and frequency details are shown in table 3.View Large Image Figure ViewerDownload Hi-res image Download (PPT)Table 4The Complete List of the 30-kb Haplotype Frequencies in 37 PopulationsPopulation Frequency (SD)HaplotypeaThe fourth position in each haplotype is the common LNP/LP variant, C/T-13910, where 1 denotes C-13910 and 2 denotes T-13910. The seventh position denotes the SNP G/A-22018, where 1 denotes G-22018 and 2 denotes A-22018.SKo (N=46)Han (N=200)ObU (N=124)Kom (N=20)Udm (N=60)Mok (N=60)Erz (N=60)Saa (N=60)FiW (N=306)FiE (N=154)DaD (N=34)DaN (N=40)DaM (N=46)Bal (N=46)Bur (N=60)Kas (N=40)Kal (N=60)Pat (N=56)Haz (N=28)Blu (N=38)Sin (N=56)Bra (N=60)MaB (N=58)Moh (N=58)Par (N=58)Ira (N=42)Qas (N=20)Ara (N=102)SIt (N=200)Fra (N=34)Bas (N=170)Uta (N=184)Som (N=158)Mor (N=180)Sah (N=114)Aam (N=100)Ful (N=88)H1111111111.11 (.05).21 (.03).19 (.04).15 (.09).04 (.03).32 (.06).17 (.05).12 (.04).10 (.02).11 (.02).23 (.07).40 (.07).33 (.07).54 (.07).18 (.04).18 (.07).37 (.08).21 (.07).36 (.09).17 (.06).21 (.06).17 (.05).22 (.06).29 (.07).39 (.07).38 (.09).60 (.12).41 (.05).32 (.04).34 (.09).12 (.03).10 (.02).27 (.04).31 (.03).25 (.05).19 (.04).01 (.01)H2111111112.35 (.08).12 (.02).31 (.04).10 (.08).19 (.06).18 (.05).25 (.07).25 (.05).12 (.02).16 (.03).50 (.09).15 (.05).26 (.07).13 (.07).44 (.07).31 (.08).22 (.05).27 (.06).11 (.06).33 (.07).23 (.06).33 (.06).30 (.06).31 (.06).21 (.06).26 (.08).30 (.11).26 (.05).21 (.03).13 (.06).09 (.02).05 (.02).38 (.04).20 (.03).34 (.05).17 (.04).07 (.04)H3111111121.01 (.01).01 (.01).01 (.01)H4111111122.26 (.08).28 (.03).02 (.01).25 (.12).07 (.04).07 (.04).03 (.01).05 (.01).03 (.03).05 (.04).07 (.04).04 (.03).03 (.03).15 (.05).07 (.03).18 (.08).03 (.02).02 (.02).03 (.02).13 (.04).12 (.04).02 (.02).03 (.01).06 (.05).01 (.01).02 (.01).04 (.02).06 (.02).01 (.01).32 (.05).05 (.02)H5111111211.02 (.02).01 (.01).01 (.01)H6111111212.02 (.02).05 (.03).01 (.01).02 (.01)H7111111222.02 (.02).01 (.01)H8111211111.02 (.02).05 (.03)H9111211112.02 (.02).05 (.04).01 (.01).01 (.00).01 (.01).02 (.01).01 (.01)H10111211122.02 (.01).00 (.00)H11111211211.07 (.04).11 (04).02 (.02).02 (.02)H12111211212.06 (.05)H13111211222.03 (.03)H14111212222.01 (.01)H15111222212.01 (01)H16111221111.02 (.02)H17111221122.02 (02).01 (.01)H18111121111.00 (.00).02 (.02).02 (.01)H19111121112.00 (.00).01 (.01).01 (.01).01 (.01).01 (.01).02 (.01)H20111121122.00 (.00).01 (.01).01 (.00)H21111122111.00 (.00)H22111112111.01 (.01).01 (.01)H23111112112.01 (.01)H24121111111.02 (.02).02 (.02.01 (.00).01 (.01)H25121111121.02 (.01)H26121111112.01 (.00).02 (.01).01 (.01)H27121111122.07 (.03).04 (.02).02 (.01).01 (.01).05 (02)H28121111222.01 (.01)H29121122122.01 (.01)H30121122211.01 (.01)H31121211112.01 (.01)H32121211122.01 (.01)H33121222222.01 (.01)H34112111111.02 (.01).02 (.02).06 (.02).01 (.01).01 (.01).02 (.01)H35112111121.02 (.02)H36112111112.02 (.02)H37112111122.01 (.01).01 (.01).01 (.00)H38112122112.01 (.01)H39112122122.01 (.01)H40112212212.01 (.01)H41112222122.01 (.01)H42111112111.01 (.01)H43122111111.00 (.00)H44122111112.02(.02).01 (.01)H45122111122.02 (.02).00 (.00).04 (.03)H46122122122.22 (.07).03 (.02).02 (.02).02 (.02).02 (.02).05 (.04).12 (.03).01 (.01)H47122122222.01 (.01)H48122222212.10 (.7)H49122222222.02 (.02).06 (.04).03 (.01).03 (.03)H50122211111.00 (.00)H51211111111.08 (.04).03 (.02).01 (.01).04 (.03).02 (.02).02 (.02).02 (.02).06 (.02).01 (.01)H52211111112.03 (.02).00 (.00).03 (.03)H53211111211.04 (.03).01 (.01).01 (.01).01 (.01).07 (.02).14 (.05)H54211111122.02 (.01).01 (.01).02 (.02).03 (.02).06 (.03)H55211111222.01 (.01)H56211121111H57211112112.00 (.00)H58211122111.01 (.01)H59211122112.00 (.00)H60211122121.03 (.03).01 (.00)H61211122122.02 (.02).02 (.01).02 (.02)H62211122212H63211211222.02 (.02).02 (.02)H64211222111.01 (.01)H65211222222.02 (.02).02 (.03)H66211211222.03 (.03).00 (.00)H67212111111.01 (.01).00 (.00).01 (.01).01 (.01)H68212122122.02 (.02).01 (.01)H69212222211.01 (.01)H70212212222.02 (.02).02 (.02).03 (.02)H71212222222.02 (.02).02 (.02)H72221122122.01 (.01)H73221122222.01 (.01)H74221222211.01 (.01)H75221222222.02 (.02)H76222111111.00 (.00).01 (.01).01 (.00)H77222111112.01 (.01)H78222112122.02 (.02).00 (.00).03 (.03)H79222121122.02 (.02).01 (.01).02 (.02).00 (.00).02 (.01)H80222121212.02 (.02)H81222121112.02 (.01).00 (.00).03 (.01).09 (.04).05 (.04).03 (.02).04 (.03).02 (.01).01 (.01).02 (.01).01 (.01).02 (.01)H82222122112.02 (.03).02 (.01).05 (.04).05 (.03).02 (.02).01 (.01)H83222122121.03 (.03).04 (.04)H84222122122.04 (.03)36 (.03).33 (.04).35 (.12).35 (.07).09 (.04).13 (.05).33 (.08).06 (.01).11 (.02).05 (.05).08 (.04).19 (.06).15 (.06).18 (.05).20 (.08).18 (.05).07 (.04).18 (.10).04 (.03).09 (.04).08 (.04).17 (.06).09 (.04).11 (.04).17 (.07).05 (.04).04 (.02).04 (.02).09 (.06).03 (.01).04 (.02).15 (.03).15 (.03).06 (.02).09 (.03).06 (.03)H85222122212.01 (.01)H86222122221.03 (.02).01 (.01)H87222122222.00 (.00).08 (.04).03 (.02).02 (.02).07 (.06).05 (.04).02 (.02).01 (.02).02 (.01).03 (.03).02 (.01).02 (.01)H88222211221.01 (.01)H89222212221.02 (.02)H90222212222.01 (.01).00 (.00).02 (.01).02 (.01)H91222221221.02 (.01)H92222221222.01 (.00)H93222221112.00 (.00).03 (.03)H94222222121.01 (.01)H95222222122.03 (.02).05 (.03).02 (.01).01 (.01).01 (.01).01 (.01)H96222222212.01 (.01).01 (.01)H97222222221.09 (.05).02 (.02).01 (.01).01 (.01)H98222222222.03 (.02).05 (.06).14 (.05).14 (.05).20 (.06).12 (.04).61 (03).53 (.04).13 (.06).11 (.05).06 (.05).27 (.06).04 (.03).34 (.08).39 (.07).25 (.06).15 (.05).28 (.06).01 (.01).02 (.02).05 (.05).05 (.02).01 (.01).31 (.09).56 (.04).74 (.03).01 (.01).12 (.02).26 (.04).06 (.03).47 (.06)H103212112110.02 (.02)H104221221220.02 (.02)Note.—Haplotypes >4% in any population have been included in the table. Alleles coded as 1 are based on the sequence of the primate samples and were cosegregated with the LNP phenotype, and alleles coded as 2 represent the alleles cosegregated with the LP phenotype. The SNPs used in constructing the haplotypes are shown in table 2. The three-letter codes for the populations are used; the complete names are given in table 3.a The fourth position in each haplotype is the common LNP/LP variant, C/T-13910, where 1 denotes C-13910 and 2 denotes T-13910. The seventh position denotes the SNP G/A-22018, where 1 denotes G-22018 and 2 denotes A-22018. Open table in a new tab Table 5A List of the Population Haplotype Frequencies Depicted in the MJ Network of Figure 4Population Frequency (% [SD])HaplotypeSKoHanObUKomUdmMokErzSaaFiWFiEDaDDaNDaMBalBurKasKalPatHazN462001242060606060154306344046466040605628LNP: H111 (5)21 (3)19 (4)15 (9)4 (3)32 (6)17 (5)12 (4)10 (2)11 (2)23 (7)40 (7)33 (7)54 (7)18 (4)18 (7)37 (8)21 (7)36 (9) H235 (8)12 (2)31 (4)10 (8)19 (6)18 (5)25 (7)25 (5)12 (2)16 (2)50 (9)15 (5)26 (7)13 (7)44 (7)31 (8)22 (5)27 (6)11 (6) H426 (8)28 (3)2 (1)25 (12)…7 (4)…7 (4)3 (1)5 (1)3 (3)5 (4)7 (4)…4 (3)3 (3)15 (5)7 (3)18 (8) H27…………………………………7 (3)…………… H34………………………………………………… H4622 (7)…3 (2)……2 (2)2 (2)2 (2)…………………5 (4)……… H51………………8 (4)3 (2)1 (1)………………4 (3)……… H52……3 (2)……………0 (0)…………2 (2)…………4 (3) H54…2 (1)1 (1)…………2 (2)……………4 (3)…………… H55………………………………………3 (2)……… H81………………2 (1)…0 (0)3 (1)…………9 (4)5 (4)3 (2)…4 (3) H822 (3)…2 (1)……………………5 (4)5 (3)2 (2)…………… H844 (3)36 (3)33 (4)35 (12)35 (7)9 (4)13 (5)33 (8)6 (1)11 (2)5 (5)8 (4)19 (6)15 (6)18 (5)20 (8)18 (5)7 (4)18 (10) H87……………………0 (0)……8 (4)…………3 (2)2 (2)7 (6)LP: H8…………2 (2)…5 (3)……………………………… H9………………………………………………… H11…………7 (4)11 (4)………………………………… H12…………6 (5)…………………………………… H48………10 (7)……………………………………… H49…………………2 (2)…………………6 (4)……… H95…………………3 (2)…………………………… H97…………………………9 (5)………2 (2)1 (1)……… H98……3 (2)5 (6)14 (5)14 (5)20 (6)12 (4)61 (4)53 (3)…13 (6)11 (5)……6 (5)…27 (6)4 (3)Note.—Haplotypes presented are restricted to the haplotypes with frequency >4% in any of the populations analyzed. The SNPs used for haplotype construction are SNPs 1–9 (table 1). The three-letter codes for the populations are used; the complete names are given in table 3. Open table in a new tab Note.— Haplotypes >4% in any population have been included in the table. Alleles coded as 1 are based on the sequence of the primate samples and were cosegregated with the LNP phenotype, and alleles coded as 2 represent the alleles cosegregated with the LP phenotype. The SNPs used in constructing the haplotypes are shown in table 2. The three-letter codes for the populations are used; the complete names are given in table 3. Note.— Haplotypes presented are restricted to the haplotypes with frequency >4% in any of the populations analyzed. The SNPs used for haplotype construction are SNPs 1–9 (table 1). The three-letter codes for the populations are used; the complete names are given in table 3. One of the ni

Serial monitoring data for faecal calprotectin and lactoferrin during Crohn's disease (CD) therapy are scarce. The aim of this research was to study the behaviour of faecal biomarkers during CD therapy.Adult CD patients (n = 19) needing therapy enhancement were prospectively recruited. The simple endoscopic score for Crohn's disease (SES-CD) was administered before and 4-6 months after therapy. At baseline and at 2-3 and 4-6 months, patients provided faecal samples for measurements of calprotectin and lactoferrin.Of 19 patients, seven were endoscopic responders, three were partial responders and nine were non-responders. During therapy, both faecal-biomarker concentrations decreased significantly in responders: median calprotectin from 1282 microg/g (range 156-2277 microg/g) to 73 microg/g (range 7-2222; P = 0.005) and lactoferrin from 233 microg/g (range 2.8-802 microg/g) to 0.0 microg/g (range 0.0-420 microg/g; P = 0.005), and these changes correlated significantly with changes in the SES-CD. In non-responders, changes in faecal biomarkers were non-significant: calprotectin decreased from 1017 microg/g (range 53-3928 microg/g) to 223 microg/g (range 35-15330 microg/g; P = 0.594) and lactoferrin from 22.5 microg/g (range 2.1-629 microg/g) to 13.0 microg/g (range 3.5-1259 microg/g; P = 0.515).The faecal neutrophil-derived proteins calprotectin and lactoferrin are reliable surrogate markers of mucosal improvement. Endoscopic responders achieved normalization of faecal biomarkers, whereas in the majority of endoscopic non-responders these markers remained abnormal.

The disease process leading to clinical type 1 diabetes often starts during the first years of life. Early exposure to complex dietary proteins may increase the risk of β-cell autoimmunity in children at genetic risk for type 1 diabetes. Extensively hydrolyzed formulas do not contain intact proteins.To test the hypothesis that weaning to an extensively hydrolyzed formula decreases the cumulative incidence of diabetes-associated autoantibodies in young children.A double-blind randomized clinical trial of 2159 infants with HLA-conferred disease susceptibility and a first-degree relative with type 1 diabetes recruited from May 2002 to January 2007 in 78 study centers in 15 countries; 1078 were randomized to be weaned to the extensively hydrolyzed casein formula and 1081 were randomized to be weaned to a conventional cows' milk-based formula. The participants were observed to April 16, 2013.The participants received either a casein hydrolysate or a conventional cows' milk formula supplemented with 20% of the casein hydrolysate.AND MEASURES: Primary outcome was positivity for at least 2 diabetes-associated autoantibodies out of 4 analyzed. Autoantibodies to insulin, glutamic acid decarboxylase, and the insulinoma-associated-2 (IA-2) molecule were analyzed using radiobinding assays and islet cell antibodies with immunofluorescence during a median observation period of 7.0 years (mean, 6.3 years).The absolute risk of positivity for 2 or more islet autoantibodies was 13.4% among those randomized to the casein hydrolysate formula (n = 139) vs 11.4% among those randomized to the conventional formula (n = 117). The unadjusted hazard ratio for positivity for 2 or more autoantibodies among those randomized to be weaned to the casein hydrolysate was 1.21 (95% CI, 0.94-1.54), compared with those randomized to the conventional formula, while the hazard ratio adjusted for HLA risk, duration of breastfeeding, vitamin D use, study formula duration and consumption, and region was 1.23 (95% CI, 0.96-1.58). There were no clinically significant differences in the rate of reported adverse events between the 2 groups.Among infants at risk for type 1 diabetes, the use of a hydrolyzed formula, when compared with a conventional formula, did not reduce the incidence of diabetes-associated autoantibodies after 7 years. These findings do not support a benefit from hydrolyzed formula. TRIAL REGISTRATION clinicaltrials.gov Identifier: NCT00179777.

BackgroundThe dynamics and balance of allergen-specific IgE, IgG4, and IgA binding might contribute to the development of tolerance in patients with cow's milk allergy (CMA). Profiling of antibody binding to cow's milk (CM) protein epitopes might help in predicting the natural history of allergy.ObjectiveWe sought to investigate differences in IgE, IgG4, and IgA binding to CM epitopes over time between patients with early recovery or with persisting CMA.MethodsWe studied serum samples at the time of diagnosis (mean age, 7 months), 1 year later, and at follow-up (mean age, 8.6 years) from 11 patients with persisting IgE-mediated CMA at age 8 to 9 years and 12 patients who recovered by age 3 years. We measured the binding of IgE, IgG4, and IgA antibodies to sequential epitopes derived from 5 major CM proteins with a peptide microarray–based immunoassay. We analyzed the data with a novel image-processing method together with machine learning prediction.ResultsIgE epitope–binding patterns were stable over time in patients with persisting CMA, whereas binding decreased in patients who recovered early. Binding patterns of IgE and IgG4 overlapped. Among patients who recovered early, the signal of IgG4 binding increased and that of IgE decreased over time. IgE and IgG4 binding to a panel of αs1-, αs2-, β-, and κ-casein regions predicted outcome with significant accuracy.ConclusionsAttaining tolerance to CM is associated with decreased epitope binding by IgE and a concurrent increase in corresponding epitope binding by IgG4. The dynamics and balance of allergen-specific IgE, IgG4, and IgA binding might contribute to the development of tolerance in patients with cow's milk allergy (CMA). Profiling of antibody binding to cow's milk (CM) protein epitopes might help in predicting the natural history of allergy. We sought to investigate differences in IgE, IgG4, and IgA binding to CM epitopes over time between patients with early recovery or with persisting CMA. We studied serum samples at the time of diagnosis (mean age, 7 months), 1 year later, and at follow-up (mean age, 8.6 years) from 11 patients with persisting IgE-mediated CMA at age 8 to 9 years and 12 patients who recovered by age 3 years. We measured the binding of IgE, IgG4, and IgA antibodies to sequential epitopes derived from 5 major CM proteins with a peptide microarray–based immunoassay. We analyzed the data with a novel image-processing method together with machine learning prediction. IgE epitope–binding patterns were stable over time in patients with persisting CMA, whereas binding decreased in patients who recovered early. Binding patterns of IgE and IgG4 overlapped. Among patients who recovered early, the signal of IgG4 binding increased and that of IgE decreased over time. IgE and IgG4 binding to a panel of αs1-, αs2-, β-, and κ-casein regions predicted outcome with significant accuracy. Attaining tolerance to CM is associated with decreased epitope binding by IgE and a concurrent increase in corresponding epitope binding by IgG4.

In 46 intestinal specimens from infants 2 hr to 6 months old, the numbers of immunoglobulin-containing cells were counted by the direct immunofluorescent or direct immunoperoxidase technique. A restudy was made of biopsy specimens taken for diagnostic purposes, 34 from the rectum and 3 from the transverse colon of 30 infants at the Children's Hospital, Helsinki in 1971-1977, and in addition, of autopsy material recently collected from the intestines of 9 infants. Up to the age of 12 days, no immunoglobulin-containing cells were seen. A small number (1 to 2) of IgM-containing cells was seen in the lymph nodes of the 2 earlier specimens taken at the ages of 6 and 12 days. A rectal specimen from a 12-day-old infant showed 24 IgA- and 60 IgM-containing cells/mm2. In rectal specimens of infants less than 1 month old, the mean number of IgM-containing cells (26/mm2) was higher than that of IgA-containing cells (14/mm2), but older infants had a significantly higher mean number of IgA-containing cells (P < 0.01). The mean number of IgM-containing cells was the same in children 1 to 3 months (53 cells/mm2) and 3 to 6 months of age (59 cells/mm2), whereas the mean number of IgA-containing cells increased with age up to 6 months (112 and 163 cells/mm2). The youngest infant who had IgG-containing cells was 13 days old, although positive staining of intercellular spaces in the lamina propria and of the capillary endothelium by anti-IgG serum was observed in all specimens. The mean number of IgG-containing cells was low (5 cells/mm2) in all age groups. Sparse IgE-containing cells (less than 12 /mm2) were seen in 4 of the 46 specimens. In 5 patients, 2 or 3 consecutive specimens were available for the study; in these, the increase in the numbers of IgA- and IgM-containing cells was similar to the mean increase in cell numbers for the series as a whole.

Development of oral tolerance and its stimulation by probiotics are still incomprehensible. Microbial stimulation of the gut may induce a subtle inflammation and induce secretion of mucosal IgA, which participates in antigen elimination. In a cohort of allergy-prone infants receiving probiotics and prebiotics or placebo we studied intestinal IgA and inflammation in the development of eczema, food allergy, asthma, and rhinitis (allergic diseases). We performed a nested unmatched case-control study of 237 infants participating in a randomized double-blind placebo-controlled allergy-prevention trial using a combination of four probiotic strains pre-natally and during 6 months form birth. We measured faecal IgA, alpha1-antitrypsin (alpha1-AT), tumour necrosis factor-alpha (TNF-alpha), and calprotectin at the age of 3 and 6 months. By age 2 yr, 124 infants had developed allergic disease or IgE-sensitization (cases) and 113 had not (controls). In infants with high faecal IgA concentration at the age of 6 months, the risk of having any allergic disease before the age of 2 yr tended to reduce [odds ratio (OR: 0.52)] and the risk for any IgE-associated (atopic) disease reduced significantly (OR: 0.49). High faecal calprotectin at the age of 6 months associated also with lower risk for IgE-associated diseases up to age 2 yr (OR: 0.49). All faecal inflammation markers (alpha1-AT, TNF-alpha, and calprotectin) correlated positively with faecal IgA (p < 0.001). Probiotics tended to augment faecal IgA (p = 0.085) and significantly increased faecal alpha1-AT (p = 0.001). High intestinal IgA in early life associates with minimal intestinal inflammation and indicates reduced risk for IgE-associated allergic diseases.

To investigate if there is a reduced risk of type 1 diabetes in children breastfed or exclusively breastfed by performing a pooled analysis with adjustment for recognized confounders.Relevant studies were identified from literature searches using MEDLINE, Web of Science, and EMBASE. Authors of relevant studies were asked to provide individual participant data or conduct prespecified analyses. Meta-analysis techniques were used to combine odds ratios (ORs) and investigate heterogeneity between studies.Data were available from 43 studies including 9,874 patients with type 1 diabetes. Overall, there was a reduction in the risk of diabetes after exclusive breast-feeding for >2 weeks (20 studies; OR = 0.75, 95% CI 0.64-0.88), the association after exclusive breast-feeding for >3 months was weaker (30 studies; OR = 0.87, 95% CI 0.75-1.00), and no association was observed after (nonexclusive) breast-feeding for >2 weeks (28 studies; OR = 0.93, 95% CI 0.81-1.07) or >3 months (29 studies; OR = 0.88, 95% CI 0.78-1.00). These associations were all subject to marked heterogeneity (I(2) = 58, 76, 54, and 68%, respectively). In studies with lower risk of bias, the reduced risk after exclusive breast-feeding for >2 weeks remained (12 studies; OR = 0.86, 95% CI 0.75-0.99), and heterogeneity was reduced (I(2) = 0%). Adjustments for potential confounders altered these estimates very little.The pooled analysis suggests weak protective associations between exclusive breast-feeding and type 1 diabetes risk. However, these findings are difficult to interpret because of the marked variation in effect and possible biases (particularly recall bias) inherent in the included studies.

Early exposure to complex dietary proteins may increase the risk of type 1 diabetes in children with genetic disease susceptibility. There are no intact proteins in extensively hydrolyzed formulas.To test the hypothesis that weaning to an extensively hydrolyzed formula decreases the cumulative incidence of type 1 diabetes in young children.An international double-blind randomized clinical trial of 2159 infants with human leukocyte antigen-conferred disease susceptibility and a first-degree relative with type 1 diabetes recruited from May 2002 to January 2007 in 78 study centers in 15 countries; 1081 were randomized to be weaned to the extensively hydrolyzed casein formula and 1078 to a conventional formula. The follow-up of the participants ended on February 28, 2017.The participants received either a casein hydrolysate or a conventional adapted cow's milk formula supplemented with 20% of the casein hydrolysate. The minimum duration of study formula exposure was 60 days by 6 to 8 months of age.Primary outcome was type 1 diabetes diagnosed according to World Health Organization criteria. Secondary outcomes included age at diabetes diagnosis and safety (adverse events).Among 2159 newborn infants (1021 female [47.3%]) who were randomized, 1744 (80.8%) completed the trial. The participants were observed for a median of 11.5 years (quartile [Q] 1-Q3, 10.2-12.8). The absolute risk of type 1 diabetes was 8.4% among those randomized to the casein hydrolysate (n = 91) vs 7.6% among those randomized to the conventional formula (n = 82) (difference, 0.8% [95% CI, -1.6% to 3.2%]). The hazard ratio for type 1 diabetes adjusted for human leukocyte antigen risk group, duration of breastfeeding, duration of study formula consumption, sex, and region while treating study center as a random effect was 1.1 (95% CI, 0.8 to 1.5; P = .46). The median age at diagnosis of type 1 diabetes was similar in the 2 groups (6.0 years [Q1-Q3, 3.1-8.9] vs 5.8 years [Q1-Q3, 2.6-9.1]; difference, 0.2 years [95% CI, -0.9 to 1.2]). Upper respiratory infections were the most common adverse event reported (frequency, 0.48 events/year in the hydrolysate group and 0.50 events/year in the control group).Among infants at risk for type 1 diabetes, weaning to a hydrolyzed formula compared with a conventional formula did not reduce the cumulative incidence of type 1 diabetes after median follow-up for 11.5 years. These findings do not support a need to revise the dietary recommendations for infants at risk for type 1 diabetes.clinicaltrials.gov Identifier: NCT00179777.

Children with high body mass index (BMI) at preschool age are at risk of developing obesity. Early identification of factors that increase the risk of excessive weight gain could help direct preventive actions. The intestinal microbiota and antibiotic use have been identified as potential modulators of early metabolic programming and weight development. To test if the early microbiota composition is associated with later BMI, and if antibiotic use modifies this association, we analysed the faecal microbiota composition at 3 months and the BMI at 5-6 years in two cohorts of healthy children born vaginally at term in the Netherlands (N = 87) and Finland (N = 75). We obtained lifetime antibiotic use records and measured weight and height of all children.The relative abundance of streptococci was positively and the relative abundance of bifidobacteria negatively associated with the BMI outcome. The association was especially strong among children with a history of antibiotic use. Bacteroides relative abundance was associated with BMI only in the children with minimal lifetime antibiotic exposure.The intestinal microbiota of infants are predictive of later BMI and may serve as an early indicator of obesity risk. Bifidobacteria and streptococci, which are indicators of microbiota maturation in infants, are likely candidates for metabolic programming of infants, and their influence on BMI appears to depend on later antibiotic use.

Abstract One of the most abundant components in human milk is formed by oligosaccharides, which are poorly digested by the infant. The oligosaccharide composition of breast milk varies between mothers, and is dependent on maternal secretor (FUT2) genotype. Secretor mothers produce milk containing α1-2 fucosylated human milk oligosaccharides, which are absent in the milk of non-secretor mothers. Several strains of bacteria in the infant gut have the capacity to utilise human milk oligosaccharides (HMOs). Here we investigate the differences in infant gut microbiota composition between secretor (N = 76) and non-secretor (N = 15) mothers, taking into account birth mode. In the vaginally born infants, maternal secretor status was not associated with microbiota composition. In the caesarean-born, however, many of the caesarean-associated microbiota patterns were more pronounced among the infants of non-secretor mothers compared to those of secretor mothers. Particularly bifidobacteria were strongly depleted and enterococci increased among the caesarean-born infants of non-secretor mothers. Furthermore, Akkermansia was increased in the section-born infants of secretor mothers, supporting the suggestion that this organism may degrade HMOs. The results indicate that maternal secretor status may be particularly influential in infants with compromised microbiota development, and that these infants could benefit from corrective supplementation.

Manifestation of allergic disease depends on genetic predisposition, diet and commensal microbiota. Genetic polymorphism of mothers determines their breast milk glycan composition. One major determinant is the fucosyltransferase 2 (FUT2, secretor gene) that was shown to be linked to commensal microbiota establishment. We studied whether FUT2-dependent breast milk oligosaccharides are associated with allergic disease in breast-fed infants later in life.We analyzed FUT2-dependent oligosaccharides in breast milk samples of mothers (n = 266) from the placebo group of a randomized placebo-controlled trial of prebiotics and probiotics as preventive against allergic disease in infants with high allergy risk (trial registry number: NCT00298337). Using logistic regression models, we studied associations between FUT2-dependent breast milk oligosaccharides and incidence of allergic disease at 2 and 5 years of age.At 2 years, but not at 5 years of age, we observed a presumed lower incidence (p < 0.1) for IgE-associated eczema manifestation in C-section-born infants who were fed breast milk containing FUT2-dependent oligosaccharides. By logistic regression, we observed a similar relation (p < 0.1) between presence of FUT2-dependent breast milk oligosaccharides and IgE-associated disease and IgE-associated eczema in C-section-born infants only. When testing with the levels of breast milk oligosaccharide 2'-fucosyllactose as proxy for FUT2 activity, we observed significant (p < 0.05) associations in the C-section-born infants with 'any allergic disease,' IgE-associated disease, eczema and IgE-associated eczema.The data indicate that infants born by C-section and having a high hereditary risk for allergies might have a lower risk to manifest IgE-associated eczema at 2 years, but not 5 years of age, when fed breast milk with FUT2-dependent milk oligosaccharides. Further studies with larger cohorts and especially randomized controlled intervention trials are required to build on these preliminary observations.

Intestinal failure (IF)-associated liver disease (IFALD) is the major cause of mortality in IF. The link between intestinal microbiota and IFALD is unclear.We compared intestinal microbiota of patients with IF (n = 23) with healthy controls (n = 58) using culture-independent phylogenetic microarray analysis. The microbiota was related to histological liver injury, fecal markers of intestinal inflammation, matrix metalloproteinase 9 and calprotectin, and disease characteristics.Overabundance of Lactobacilli, Proteobacteria, and Actinobacteria was observed in IF, whereas bacteria related to Clostridium clusters III, IV, and XIVa along with overall diversity and richness were reduced. Patients were segregated into 3 subgroups based on dominating bacteria: Clostridium cluster XIVa, Proteobacteria, and bacteria related to Lactobacillus plantarum. In addition to liver steatosis and fibrosis, Proteobacteria were associated with prolonged current parenteral nutrition (PN) as well as liver and intestinal inflammation. Lactobacilli were related to advanced steatosis and fibrosis mostly after weaning off PN without associated inflammation. In multivariate permutational analysis of variance, liver steatosis, bowel length, PN calories, and antibiotic treatment best explained the microbiota variation among patients with IF.Intestinal microbiota composition was associated with liver steatosis in IF and better predicted steatosis than duration of PN or length of the remaining intestine. Our results may be explained by a model in which steatosis is initiated during PN in response to proinflammatory lipopolysaccharides produced by Proteobacteria and progresses after weaning off PN, as the L plantarum group Lactobacilli becomes dominant and affects lipid metabolism by altering bile acid signaling.

Abstract Specific Ab-secreting cells (ASC) appear in the human blood as a response to oral and parenteral vaccination. The actual contribution of these cells to the defense of the body depends on their final effector site. The homing potentials of mucosally and parenterally induced ASC were compared by examining the homing receptor (HR) expression of circulating specific ASC in the blood of volunteers vaccinated orally or parenterally with the same Ag, Salmonella typhi Ty21a. Circulating lymphocytes were separated into receptor-positive and -negative populations, and the numbers of specific ASC were assayed. The alpha4 beta7 integrin, which acts as a gut HR, was expressed on all (99%) of the mucosally activated ASC, but on only 58% of the parenterally induced ASC or 58% of all Ig-secreting cells of the unvaccinated controls. L-selectin, the peripheral lymph node HR, showed an inverse distribution; it was found on 42% of mucosally activated ASC and on 86% of parenterally induced ASC. These results reveal that all of the circulating ASC after oral vaccination are committed to migrate to the mucosal compartment of the immune system, strongly arguing for a recirculation of activated mucosal cells in humans. By contrast, ASC induced by parenteral vaccination with the same Ag are mostly directed to the systemic compartment, yet a part of them has mucosal homing attitudes as well. These differences indicate that the site of Ag encounter determines the homing potential of the cell.

We aimed to assess the feasibility of a dietary intervention trial with weaning to hydrolysed formula in infants at increased risk of type 1 diabetes and to study the effect of the intervention on the emergence of diabetes-associated autoantibodies in early childhood.We studied 242 newborn infants who had a first-degree relative with type 1 diabetes and carried risk-associated HLA-DQB1 alleles. After exclusive breastfeeding, the infants underwent a double-blind, randomised pilot trial of either casein hydrolysate (Nutramigen; Mead Johnson) or conventional cow's milk-based formula until the age of 6-8 months. During a mean observation period of 4.7 years, autoantibodies to insulin, anti-glutamic acid decarboxylase and insulinoma-associated antigen-2 were measured by radiobinding assays, and islet cell antibodies (ICA) by immunofluorescence.The feasibility of screening and identifying a cohort of first-degree relatives with HLA-conferred disease susceptibility, enrolling them in a dietary intervention trial and following them for seroconversion to autoantibody positivity is established. The cumulative incidence of autoantibodies was somewhat smaller in the casein hydrolysate vs control formula group, suggesting the need for a larger well-powered study. After adjustment for duration of study formula feeding, life-table analysis showed a significant protection by the intervention from positivity for ICA (p=0.02) and at least one autoantibody (p=0.03).The present study provides the first evidence ever in man, despite its limited power, that it may be possible to manipulate spontaneous beta cell autoimmunity by dietary intervention in infancy.

Background: Large-scale screening for coeliac disease has suggested that the disease is more prevalent than anticipated. In the screening studies published, only a minor proportion of those with a positive result have undergone jejunal biopsy to confirm the diagnosis. Our aim was to search for previously undiagnosed patients with coeliac disease by means of antiendomysium antibodies, which are more specific for the disease than serum antigliadin antibodies, and to study jejunal histology in each with a positive titre. Methods: Serum from 1070 adults working at Helsinki University Central Hospital were screened for untreated coeliac disease with IgA antiendomysium antibodies. All adults with positive titres underwent jejunal biopsy for villous structure analysis and counting of CD3-positive cells and cells bearing the gamma/delta T-cell receptor. Results: Coeliac disease was confirmed in a jejunal biopsy specimen from 8 of the 11 subjects with positive antiendomysium titres-that is, a frequency of 1 in 130. Seven of these eight coeliac patients had had minor abdominal discomfort for years, and one patient had a diagnosis of hyperthyroidism. None of the patients had osteoporosis, four had low iron storages, but only two were anaemic; no other nutritional deficiencies were found. The three other adults had a positive antiendomysium titre but a normal villous structure. One of these three was regarded as a false-positive case (titre, 1 in 5). The two other subjects (titres, 1 in 400) had increased numbers of CD3-positive T cells and gamma/delta T-cell receptor-bearing cells, suggesting a predisposition for coeliac disease. Conclusions: Undiagnosed coeliac disease is common in the adult population in Finland; in this study the prevalence was 1 in 130. Screening for coeliac disease is recommended on minor suspicion.

Jejunal biopsy was performed without selection in 110 of 201 children with insulin-dependent diabetes mellitus; serum reticulin antibody, antigiladin antibody by enzyme-linked immunosorbent assay, and serum IgA were studied in all 201 children. Seven children had severe jejunal villous atrophy, giving a prevalence of celiac disease of at least 3.5%. Of the serum tests used, antigliadin antibody with ELISA was the most sensitive. Four patients adhered to a gluten-free diet, and their jejunal structure became normal; three had subsequent gluten proyocation, and the jejunal mucosa relapsed in every one. Six had HLA-B8 and-DR3 antigens, and one had B15 and-DR4 phenotypes. In most patients, a gluten-free diet had little effect on insulin dosage, urinary excretion of glucose, or serum level of hemoglobin A1. Jejunal biopsy was performed without selection in 110 of 201 children with insulin-dependent diabetes mellitus; serum reticulin antibody, antigiladin antibody by enzyme-linked immunosorbent assay, and serum IgA were studied in all 201 children. Seven children had severe jejunal villous atrophy, giving a prevalence of celiac disease of at least 3.5%. Of the serum tests used, antigliadin antibody with ELISA was the most sensitive. Four patients adhered to a gluten-free diet, and their jejunal structure became normal; three had subsequent gluten proyocation, and the jejunal mucosa relapsed in every one. Six had HLA-B8 and-DR3 antigens, and one had B15 and-DR4 phenotypes. In most patients, a gluten-free diet had little effect on insulin dosage, urinary excretion of glucose, or serum level of hemoglobin A1.

Coeliac disease was searched for in a series of 776 children with newly diagnosed IDDM. During the follow-up of 2 to 3 years from diagnosis, reticulin and gliadin antibodies were measured, and a jejunal biopsy was performed in those cases with high levels of antibodies; 19 children were identified with coeliac disease, giving the prevalence of 2.4%. In only one case had coeliac disease been diagnosed before IDDM. Nine patients with proven coeliac disease were negative for antibodies when IDDM was diagnosed, but became positive within 24 months. All patients found to have coeliac disease were positive for IgA reticulin antibodies, but only 12 of 18 (67%) showed a high level of IgA gliadin antibodies. Of the 18 patients genotyped for HLA DR locus, 14 (78%) were positive for DR3 and 10 (56%) were positive for DR4. DQB1*0201 allele was present in 17 of 18 patients (94%). Coeliac disease in children with IDDM tends to develop soon after diabetes is diagnosed. Routine screening for coeliac disease is recommended repeatedly during the first years after the diagnosis of IDDM.

Although the proportion of γδ T-cell-receptor (TCR)-bearing intraepithelial lymphocytes is increased in the jejunum of patients with active coeliac disease, an abnormality thought to be specific among those with gluten-sensitive enteropathy, the factors influencing γδ TCR expression remain uncertain. We examined the relation between genetic factors associated with coeliac disease and intraepithelial γδ T lymphocyte distribution in both coeliac patients and their healthy first-degree relatives. 41% (45/109) of healthy relatives had an increased density of γδ T cells in their mucosa and 66% had an increased density of αβ T cells. By contrast with αβ T cells, the density of γδ cells was significantly associated with genetic markers for coeliac disease susceptiblity (DR3, DQA, and DQB). We also found a dose effect of DQA and DQB genes on the number of intraepithelial γδ T cells. An increased density of γδ T cells in normal jejunal mucosa of a healthy individual with appropriate genetic determinants might be necessary for the development of the typical lesions of coeliac disease. Routine jejunal histological studies should include γδ T-cell counts, thus allowing early detection of coeliac disease latency.

Fecal calprotectin is a promising marker for the assessment of gastrointestinal inflammation. Fecal calprotectin levels were followed-up in children with inflammatory bowel disease (IBD) who were introduced to glucocorticoid therapy. The aim of this study was to assess whether the changes in fecal calprotectin levels reflect therapeutic responses.Fecal calprotectin was measured by enzyme immunoassay in 57 children (mean age 9.8 years, range 0.9-18 years) who underwent colonoscopies (IBD n=31, non-IBD disease n=13, normal n=13) and followed-up in 15 children (mean age 13 years, range 3.6-18 years) who were introduced to glucocorticoid therapy because of active IBD at 0, 2, and 4 weeks and at 4-week intervals until one month after discontinuation of the therapy.Fecal calprotectin was <100 microg/g in 70% of the children with normal findings on colonoscopy or a non-IBD disease. Fecal calprotectin was >100 microg/g in all but one child with active IBD and in 13/15 of those children who were introduced to glucocorticoids by the clinicians. Fecal calprotectin values decreased within 4 weeks in line with clinical improvement in 7 children and normalized in 4/15 children during the follow-up. Fecal calprotectin increased in 5/8 of the non-steroid-dependent children after discontinuation of glucocorticoids.Fecal calprotectin is a sensitive marker for chronic colitis. In active disease treated with glucocorticoids, fecal calprotectin levels declined in line with the clinical improvement but seldom fell within the normal range, which suggests ongoing inflammation in a clinically silent disease. The measurement of fecal calprotectin may provide new tools for the assessment of the level of gut inflammation in children with chronic colitis in the follow-up of clinical responses.

There are at least 20 rare autosomal recessive disorders that are excessively common in Finland of which congenital lactase deficiency is one. During the last 17 years we have found 16 cases. In each case the mother noted watery diarrhoea, generally after the first feed of breast milk, and at the latest, by age 10 days. The lactose malabsorption was verified at a mean age of 36 (range 3-90) days, by which time the infants were dehydrated and 15 of them weighed less than at birth (mean weight for age was -2.8 SDs). On a lactose-free elimination diet (a group of 6 on Nutramigen and a group of 10 on soy-based formula) the children caught up in growth. One infant in each group showed allergic symptoms. While the infants were being breast fed their faeces contained 20 to 80 g/l lactose. In 24 peroral lactose tolerance tests, the greatest rise in blood glucose concentration was 0.8 mmol/l. Only 2 patients showed abnormal absorption when tested within a week of lactose elimination, and in each absorption tests became normal during the elimination period. Slight to partial villous atrophy of the jejunum was present in 4 early specimens, but in later ones the mean villous height was normal. The mean height of the epithelial cells was reduced and there were fewer intraepithelial lymphocytes in patients. The lactase activities in jejunal biopsy specimens were lower than in most patients with acquired lactase deficiency, with some overlap. The maltase and sucrase activities were normal.

Enterovirus infections are frequent in infants and may cause severe complications. We set out to assess whether breastfeeding can protect against these infections and whether such an effect is related to maternal antibodies in breast milk or in the peripheral circulation of the infant.One hundred fifty infants who were prospectively followed up from birth were monitored for enterovirus infections. The duration of breastfeeding was recorded, and maternal breast milk and blood samples were regularly taken at 3-month intervals for the detection of enterovirus antibodies and RNA. Maternal serum was available from early pregnancy, delivery, and 3 months postpartum.Enterovirus infections were frequent and were diagnosed in 43% of infants before the age of 1 year and in 15% of the mothers during pregnancy. Infants exclusively breastfed for >2 weeks had fewer enterovirus infections by the age of 1 year compared with those exclusively breastfed for < or =2 weeks (0.38 vs 0.59 infections per child). High maternal antibody levels in serum and in breast milk were associated with a reduced frequency of infections. This effect was seen only in those infants breastfed >2 weeks, indicating that breast milk antibodies mediate this effect. Enterovirus RNA was not found in any of the breast milk samples.These results suggest that breastfeeding has a protective effect against enterovirus infections in infancy. This effect seems to be mediated primarily by maternal antibodies in breast milk.

Development of humoral and cellular immune responses to orally administered antigens in human beings is poorly understood, although antigen administration has been suggested as a treatment for hypersensitivity disorders and autoimmune diseases.The purpose of the study was to investigate the development of systemic immune response in infants fed with formula containing whole cow's milk proteins or hydrolyzed formula containing casein peptides.In a double-blind trial, 10 infants received cow's milk-based formula, and 10 infants received a casein hydrolysate formula until the age of 9 months. Blood samples were taken at the ages of 6, 9, and 12 months. Cellular responses were assessed by proliferation assay of peripheral blood mononuclear cells to cow's milk proteins (beta-lactoglobulin, bovine serum albumin, and alpha-casein). Humoral responses to the same proteins were measured by ELISA for IgG antibodies.Feeding infants with cow's milk-based formula induced systemic humoral and cellular responses to cow's milk proteins. T-cell response later declined, supporting the concept of oral tolerization. Exposure to cow's milk proteins after the age of 9 months resulted in depressed cellular and humoral responsiveness to these proteins.Our results support the view that induction of oral tolerance in human beings is an age-dependent phenomenon.

Associations between infant-feeding patterns and risk of IDDM were investigated in a nationwide Finnish case-control study of 690 IDDM children < 15 yr of age. Each child was matched by date of birth and sex to a randomly selected population-based control child. Univariate analysis revealed that the risk of IDDM was increased by approximately 1.5 in children for whom breast-feeding was terminated at < 2 mo of age, doubled in those who were exclusively breast-fed for < 2 mo, and doubled in those who were introduced to dairy products at < 2 mo of age. In further multivariate analyses of these factors, it was found that introduction of dairy products at an early age was the most important risk factor, and the observed univariate effects of duration of breast-feeding variables were explained by their correlation with this factor. This is the first observational study to show that early introduction of dairy products is independently associated with an increased risk of IDDM. Adjustment for mother's education and age, child's birth order, or birth weight did not affect the results.

The densities of T cells and of cells bearing the T cell receptors gamma/delta and alpha/delta and the surface antigens CD4 and CD8 in jejunal specimens from 21 patients with dermatitis herpetiformis were compared with those in specimens from 13 untreated adults with coeliac disease and 13 control subjects. In the lamina propria of the jejunum the median density of gamma/delta+ cells was significantly (p less than 0.001) greater in untreated patients with dermatitis herpetiformis than in control subjects (114 v 36 cells/mm2) and similar to that found in the patients with coeliac disease (115 cells/mm2). The difference in gamma/delta+ cell density between patients with dermatitis herpetiformis and control subjects was much greater in the surface epithelium of the jejunum: the median density for 14 untreated patients with dermatitis herpetiformis was 39 cells/mm, for seven patients with dermatitis herpetiformis on a gluten free diet 34 cells/mm, and for control subjects 2 cells/mm; the coeliac patients had the same density as the patients with dermatitis herpetiformis (45 cells/mm). The higher density of cells bearing the alpha/delta T cell receptor in the epithelium (median 77 cells/mm) of untreated patients with dermatitis herpetiformis was associated with a gluten containing diet; in specimens taken from patients with dermatitis herpetiformis on a gluten free diet the median density was similar to that in the control subjects (44 v 39 cells/mm). The increase in the number of lymphocytes bearing the T cell receptor gamma/delta, particularly in the epithelium of the jejunum, seems to be a constant marker for these closely related diseases, whereas the density of alpha/delta+ T cells is dependent on the diet.

Fifty seven children with dermatitis herpetiformis, 18 from Finland and 39 from Hungary, were studied.Diagnostic criteria included the finding of granular IgA deposits in the skin of all patients.The mean age at onset of the rash was 7-2 years and favoured sites were the elbows, knees, and buttocks.Symptoms suggesting small intestinal disease were rare but in 35 (61%) of the children subtotal villous atrophy and in 16 (28%) partial villous atrophy were found on jejunal biopsy.Eighteen children underwent a second biopsy after a mean of 21 months on a gluten free diet; villous height was found to be increased and the intraepithelial lymphocyte count decreased in all these patients.Gluten challenge caused a reversal in the two children who underwent a third biopsy.The effect of the gluten free diet on the rash was examined in Finnish children by observing the daily requirements of dapsone, a drug used to control the rash at the beginning of the diet.Eight (67%) of the 12 children were able to stop taking dapsone after a mean of 11 months on the diet and all three patients treated with diet alone became asymptomatic after three to 6 months on the diet.These results confirm that most children with dermatitis herpetiformis have jejunal villous atrophy, though they rarely have gastrointestinal symptoms.The central role of gluten in childhood dermatitis herpetiformis is evidenced by the fact that a gluten free diet helps the damaged jejunal mucosa to recover and controls the rash even in those children who do not have an abnormal jejunal biopsy.

Probiotics are immunomodulatory and may thus affect vaccine antibody responses. With the accumulating evidence of their health‐promoting effects, probiotics are increasingly administered in allergy‐prone infants. Therefore, we studied the effect of probiotics on antibody responses to diphtheria, tetanus and Haemophilus influenzae type b (Hib) vaccines in 6‐month‐old infants participating in a randomized placebo‐controlled double‐blind allergy‐prevention trial. Mothers of unborn children at increased risk for atopy used a combination of four probiotic strains, or a placebo, for 4 wk before delivery. During 6 months from birth, their infants received the same probiotics and galacto‐oligosaccharides, or a placebo. The infants were immunized with a DTwP (diphtheria, tetanus and whole cell pertussis) at ages 3, 4, and 5 months, and with a Hib polysaccharide conjugate at 4 months. Serum diphtheria, tetanus, and Hib IgG antibodies were measured at 6 months. In the probiotic group, protective Hib antibody concentrations (≥1 μ g/ml) occurred more frequently, 16 of 32 (50%) vs. six of 29 (21%) (p = 0.020), and the geometric mean (inter‐quartile range) Hib IgG concentration tended to be higher 0.75 (0.15–2.71) μ g/ml than in the placebo group 0.40 (0.15–0.92) μ g/ml (p = 0.064). In these respective groups, diphtheria, 0.38 (0.14–0.78) vs. 0.47 (0.19–1.40) IU/ml (p = 0.449), and tetanus, 1.01(0.47–1.49) vs. 0.81 (0.56–1.39) IU/ml (p = 0.310), IgG titers were comparable. In conclusion, in allergy‐prone infants probiotics seem not to impair antibody responses to diphtheria, tetanus, or Hib, but may improve response to Hib immunization.

<b>Background:</b> The prevalence of lactase persistence is high in Saudi Arabia. <b>Objective:</b> To identify a DNA variant for the lactase persistence/non-persistence trait in adult Arabs in Saudi Arabia. <b>Methods:</b> We sequenced DNA from 432 anonymous neonatal blood donors from five different regions of Saudi Arabia to cover the 400 bp region surrounding the previously identified lactase persistence/non-persistence variant C/T−13910 residing in intron 13 of the <i>MCM6</i> gene. <b>Results:</b> Two anonymous blood donors carried the C/T−13910 genotype. One variant, T/G −13915, residing 5 bp upstream of the C/T−13910 variant, was present in 332 of 432 (76.9%) of the neonatal samples, compatible with previous prevalence figures of lactase persistence in urban Saudi populations. Determination of disaccharidase activities in 25 intestinal biopsy samples showed a highly significant correlation between lactase activity and the T/G−13915 genotypes (p&lt;0.001; Fisher exact test) as well as between the L:S ratio and the aforementioned genotypes (p&lt;0.001; Fisher exact test). <b>Conclusion:</b> The T/G−13915 variant is the founder mutation of lactase persistence in an urban Saudi population. The results obtained here have implications for genetic testing of adult-type hypolactasia and to analysis of human evolution, the origin of cattle domestication and migrations of the populations in the Arabian peninsula.

AllergyVolume 36, Issue 2 p. 73-88 Free Access Cow's Milk Allergy Erkki Savilahti, Corresponding Author Erkki Savilahti Children's Hospital, University of Helsinki, Helsinki, FinlandErkki Savilahti, M.D., Children's Hospital University of Helsinki SF-00290 Helsinki 29 FinlandSearch for more papers by this author Erkki Savilahti, Corresponding Author Erkki Savilahti Children's Hospital, University of Helsinki, Helsinki, FinlandErkki Savilahti, M.D., Children's Hospital University of Helsinki SF-00290 Helsinki 29 FinlandSearch for more papers by this author First published: February 1981 https://doi.org/10.1111/j.1398-9995.1981.tb04101.xCitations: 60AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat Citing Literature Volume36, Issue2February 1981Pages 73-88 ReferencesRelatedInformation

Because the role of regulatory T cells in the intestinal inflammation is unknown in coeliac disease (CD) and type 1 diabetes (T1D), the expression of forkhead box P3 (FoxP3), CD25, transforming growth factor-beta, interferon (IFN)-gamma, interleukin (IL)-4, IL-8, IL-10, IL-15 and IL-18 was measured by quantitative reverse transcription-polymerase chain reaction in the small intestinal biopsies from paediatric patients with active or potential CD, T1D and control patients. The numbers of FoxP3- and CD25-expressing cells were studied with immunohistochemistry. Enhanced intestinal expressions of FoxP3, IL-10 and IFN-gamma mRNAs were found in active CD when compared with controls (P-values < 0.001, 0.004, <0.001). In potential CD, only the expression of IFN-gamma mRNA was increased. The numbers of FoxP3-expressing cells were higher in active and potential CD (P < 0.001, P = 0.05), and the ratio of FoxP3 mRNA to the number of FoxP3-positive cells was decreased in potential CD when compared with controls (P = 0.007). The ratio of IFN-gamma to FoxP3-specific mRNA was increased in active and potential CD (P = 0.001 and P = 0.002). Patients with T1D had no changes in regulatory T cell markers, but showed increased expression of IL-18 mRNA. The impaired up-regulation of FoxP3 transcripts despite the infiltration of FoxP3-positive cells in potential CD may contribute to the persistence of inflammation. The increased ratio of IFN-gamma to FoxP3 mRNA in active and potential CD suggests an imbalance between regulatory and effector mechanisms. The increased intestinal expression of IL-18 mRNA in patients with T1D adds evidence in favour of the hypothesis that T1D is associated with derangements in the gut immune system.

A morphometric study of intraepithelial (IE) lymphocytes per 100 epithelial cells, villous heights (VH), crypt depths (CrD), and epithelial cell heights (ECH) was made on jejunal specimens of 17 patients with cow's-milk allergy (CMA), 52 with celiac disease (CD), seven with congenital lactase deficiency (CLD), four with acrodermatitis enteropathica (AE), four with giardiasis, and four with dermatitis herpetiformis (DH). The aim of this study was to investigate how the morphometric parameters correlate with each other. All cases with CMA, CD, and DH had villous atrophy with hyperplasia of the crypts, both signs being more severe in cases with CD and DH than with CMA. IE lymphocyte infiltration was more intense in specimens of patients with CD and DH (mean 76.0), than those with CMA (mean 62.5). The ECH were equally reduced in patients with CD and CMA. In a follow-up specimen at 1 year and 10 months for CD patients and 11 months for CMA patients the inflammation was reduced, and the VH were increased but still differed from the controls. In CLD cases the morphology of the villi and crypts of the jejunum was quite normal, with no IE lymphocyte infiltration; ECH were reduced. Minor morphological changes were seen in the specimens of patients with AE and giardiasis. In the whole study group there was a significant linear correlation, either positive or negative, between all variables measured (IE lymphocytes, VH, CrD, and ECH).

Background: Early exposure to cow’s milk (CM) proteins have been implicated in the pathogenesis of type 1 diabetes (T1D). Objective: We analyzed the development of the humoral immune response to dietary CM proteins in early childhood and its relation to later T1D. Subjects and methods: We studied a subgroup of 94 children randomized to be weaned to a CM-based infant formula in the trial to reduce insulin-dependent diabetes mellitus in the genetically at risk (TRIGR) pilot study. All subjects carried human leukocyte antigen-conferred T1D susceptibility and had an affected first-degree relative. After 7 years of follow-up, 8 subjects had progressed to T1D, 15 had at least one disease-associated autoantibody, and 71 remained autoantibody negative (controls). Immunoglobulin (Ig) G and IgA class antibodies to whole CM formula, beta-lactoglobulin (BLG), bovine serum albumin, and alpha-casein and IgG antibodies to bovine insulin (BI) were measured with enzyme-linked immunosorbent assays from sequential samples. Results: The children with later T1D showed increased IgG levels to BLG from 3 to 18 months of age (p = 0.028) and enhanced IgA levels to CM formula at the age of 9 months (p = 0.022) compared with controls. In the children with an affected father or sibling, IgG antibodies to BI were higher in autoantibody-positive subjects than in autoantibody-negative subjects at 18 months of age (p = 0.022). Conclusion: An enhanced humoral immune response to various CM proteins in infancy is seen in a subgroup of those children who later progress to T1D. Accordingly, a dysregulated immune response to oral antigens is an early event in the pathogenesis of T1D.

Data on fecal calprotectin and S100A12 in predicting wireless capsule endoscopy (WCE) findings in suspicion of Crohn's disease (CD) are scarce. Our aim was to study the role of calprotectin and S100A12 in predicting inflammatory lesions of small bowel in patients undergoing WCE.84 patients undergoing WCE (77 for suspicion of CD and 7 CD patients for evaluation of disease extent) were prospectively recruited. WCE findings were scored. Patients provided a stool sample for measurements of biomarkers. Patients underwent an esophagogastroduodenoscopy and ileocolonoscopy before WCE.WCE was abnormal in 35 (42%) of 84 patients: 14 patients with CD, 8 with NSAID enteropathies, 8 with angioectasias, 4 with polyps or tumors, and 1 with ischemic stricture. Median calprotectin concentration in the study population was 22 μg/g (range 2-342) and S100A12 concentration 0.048 μg/g (range 0.003-1.215). Fecal calprotectin was significantly higher in CD patients (median 91, range 2-312) compared with those with normal WCE or other abnormalities (p = 0.008), whereas fecal S100A12 (0.087 μg/g, range 0.008-0.896) did not differ between the groups (p = 0.166). In detecting inflammatory small bowel lesions, sensitivity, specificity, positive predictive value, and negative predictive value for fecal calprotectin (cutoff 50 μg/g) were 59%, 71%, 42%, and 83%, and for S100A12 (cutoff 0.06 μg/g) these were 59%, 66%, 38%, and 82%.In predicting small bowel inflammatory changes, fecal biomarkers calprotectin and S100A12 have moderate specificity, but low sensitivity. Neither fecal calprotectin nor S100A12 can be used for screening or excluding small bowel CD.

SUMMARY Three infants, in whom the malabsorption syndrome, small intestinal. mucosal damage and clinical cow's milk intolerance were found, were challenged with cow's milk after initial treatment with breast milk. The small intestinal mucosa was investigated with light and electron microscopy both before and after provocation. Clinically one patient reacted rapidly in a few hours and showed round cell infiltration in the surface epithelium and lamina propria 20 hours after a single challenge. Electron microscopy showed short microvilli, abnormal nuclei and thickened basement lamina in the surface epithelium. Two patients reacted slowly. One of them showed similar changes 4 days after commencement of the provocations, but the changes were much more evident 38 days later, when symptoms were also apparent. At this time large accumulations of lysosomes in the apical part of the surface epithelial cell and marked thickening of the basal lamina with accumulations of whirly collagen fibres were detected. The third patient reacted in a milder way, both clinically and morphologically. This study indicates that cow's milk, apparently through its protein fraction, may damage the surface epithelium of the small intestinal mucosa. These alterations during provocation periods resemble those found in coe‐liac disease during gluten provocation.

Oral immunotherapy (OIT) with cow's milk (CM) has been reported to induce a number of specific antibody responses, but these remain to be fully characterized. Our objective was to explore whether IgE and IgG4 epitope binding profiles could predict the risk of side effects during CM OIT.The study population consisted of 32 children (6-17 yr of age) with CM allergy: 26 children who successfully completed OIT and six children who discontinued therapy due to adverse reactions. We investigated sera drawn before and after OIT. We analyzed specific IgE and IgG4 binding to CM protein-derived peptides with a microarray-based immunoassay. Antibody binding affinity was analyzed with a competition assay where CM proteins in solution competed with peptides printed on the microarray.IgE binding to CM peptides decreased and IgG4 binding increased following the OIT in children who attained desensitization. Compared with children who successfully completed OIT, those who discontinued OIT due to adverse reactions developed increased quantities and affinity of epitope-specific IgE antibodies and a broader diversity of IgE and IgG4 binding, but less overlap in IgE and IgG4 binding to CM peptides.Detailed analysis of IgE and IgG4 binding to CM peptides may help in predicting whether CM OIT will be tolerated successfully. It may thus improve the safety of the therapy.

Abstract Accumulating evidence indicates that gut microbiota may regulate sex-hormone levels in the host, with effects on reproductive health. Very little is known about the development of intestinal microbiota during puberty in humans. To assess the connection between pubertal timing and fecal microbiota, and to assess how fecal microbiota develop during puberty in comparison with adult microbiota, we utilized a Finnish allergy-prevention-trial cohort (Flora). Data collected at 13-year follow-up were compared with adult data from a different Finnish cohort. Among the 13-year-old participants we collected questionnaire information, growth data from school-health-system records and fecal samples from 148 participants. Reference adult fecal samples were received from the Health and Early Life Microbiota (HELMi) cohort (n = 840). Fecal microbiota were analyzed using 16S rRNA gene amplicon sequencing; the data were correlated with pubertal timing and compared with data on adult microbiota. Probiotic intervention in the allergy-prevention-trial cohort was considered as a confounding factor only. The main outcome was composition of the microbiota in relation to pubertal timing (time to/from peak growth velocity) in both sexes separately, and similarity to adult microbiota. In girls, fecal microbiota became more adult-like with pubertal progression (p = 0.009). No such development was observed in boys (p = 0.9). Both sexes showed a trend towards increasing relative abundance of estrogen-metabolizing Clostridia and decreasing Bacteroidia with pubertal development, but this was statistically significant in girls only (p = 0.03). In girls, pubertal timing was associated positively with exposure to cephalosporins prior to the age of 10. Our data support the hypothesis that gut microbiota, particularly members of Ruminococcaceae , may affect pubertal timing, possibly via regulating host sex-hormone levels. Trial registration The registration number for the allergy-prevention-trial cohort: ClinicalTrials.gov, NCT00298337, registered 1 March 2006—Retrospectively registered, https://clinicaltrials.gov/show/NCT00298337 . The adult-comparison cohort (HELMi) is NCT03996304.

The diagnosis of cow's milk allergy is based on a clinical response to an elimination-challenge test with cow's milk. We studied the usefulness of the skin-prick and patch tests and measurement of cow's milk-specific IgE and eosinophil cationic protein in serum as diagnostic tools for cow's milk allergy in a cohort of 6209 unselected infants followed from birth for the development of cow's milk allergy. Of the 239 infants challenged with cow's milk, 118 showed a positive and 121 a negative response at a mean age of 6.9 months. A positive reaction to a skin-prick test with cow's milk (> or = 3 mm) was seen in 72 (61%) and 29 (24%) infants with positive and negative challenges, elevated serum cow's milk-specific IgE (> or = 0.7 kU/L) in 52 (45%) and 15 (13%) infants, a positive reaction to patch test with cow's milk protein fractions in 26 (26%) and eight (8%) infants, and elevated serum eosinophil cationic protein (> or = 20 microg/L) in 22 (21%) and seven (13%) infants, respectively. Parallel use of the four tests with the above-mentioned cut-off values correctly classified 73% of the infants with a sensitivity of 0.76 and a specificity of 0.67. An immediate reaction to cow's milk challenge correlated with skin prick test positivity and elevated serum milk-specific IgE, and tended to correlate with patch test positivity. No single test or parallel use of the four tests could predict the challenge outcome acceptably in this prospectively followed, unselected cohort of 6209 infants. A positive reaction to one or more tests needs to be confirmed by a challenge test and a negative response to all four tests does not rule out the possibility of cow's milk allergy.

To investigate the humoral immune response to cow's milk in pediatric patients with newly diagnosed IDDM.We measured IgA, IgG, and IgM antibodies to the proteins of cow's milk and to beta-lactoglobulin by an enzyme-linked immunosorbent assay. Samples from 706 pediatric patients with newly diagnosed IDDM were available. We used two comparison groups: 105 patients < 7 yr old had an unrelated age-matched control subject, and samples from 456 3- to 14-yr-old nondiabetic siblings also were available.Patients < 3 yr of age had a markedly higher median level of IgG and IgA antibodies to cow's milk compared with the control subjects (P = 0.03 and 0.002, respectively), IgG antibodies to beta-lactoglobulin also were higher (P = 0.03). Older groups of diabetic patients, 3.0-6.9 and 7.0-14.9 yr of age, had significantly higher levels of IgA antibodies to cow's milk and beta-lactoglobulin than the age-matched comparison groups of both unrelated control subjects and nondiabetic siblings, although the median values of the diabetic patients were closer to those of the comparison groups than in the youngest groups. Nondiabetic siblings had higher levels of IgA cow's milk antibodies than unrelated control subjects of similar ages (3-6.9 yr of age) (P = 0.03). The 14 siblings contracting IDDM during the follow-up showed no change in the levels of cow's milk or beta-lactoglobulin antibodies in relation to the clinical diagnosis.The results indicate an abnormally strong humoral response to the proteins of cow's milk, particularly in young IDDM patients and, to a lesser extent, in their siblings. We infer that the proteins of cow's milk may trigger the autoimmune process of IDDM.

A syndrome of chronic diarrhea, vomiting, and failure to thrive was described 35 years ago. The syndrome was caused by damage in the jejunum after ingestion of cow's milk. Symptoms appeared in young infants shortly after introduction of cow's milk formula. Patients had moderate steatorrhea, decreased absorption of D-xylose, and, often, iron-deficiency anemia and hypoproteinemia. They had strong IgA and IgG antibodies to cow's milk. IgE antibodies to cow's milk were negative, as a rule. Indicators of cell-mediated immune reaction to cow's milk proteins were often positive. Patients were tolerant to cow's milk by the age of 3 years. Malabsorption was due to damage to the jejunal mucosa: Varying villus atrophy was associated with inflammation in surface epithelium and lamina propria. The epithelial cell renewal rate increased. Surface epithelial cells decreased in height, with short, furry microvilli and large aggregates of lysozymes. The number of intraepithelial lymphocytes was markedly increased, but normalized during cow's milk elimination. Most of these lymphocytes had alpha/beta T-cell receptors, and many were cytotoxic. Some specimens had an increase in gamma/delta T-cell receptor-bearing cells. In the lamina propria, CD4+ cells predominated, and some of them were activated. IgA- and IgM-containing cells were markedly increased during cow's milk exposure, but IgE cells were not abnormal. The density of eosinophils was moderately increased. Secretion of interferon-gamma by cells isolated from patients' intestines was markedly increased. Morphologic and immunologic findings suggest that T-cell-mediated reaction to proteins in cow's milk is present in the small intestines of patients with this syndrome and causes this enteropathy.

Congenital lactase deficiency (CLD) is a severe gastrointestinal disorder characterized by watery diarrhea in infants fed with breast milk or other lactose-containing formulas. We initially assigned the CLD locus by linkage and linkage disequilibrium on 2q21 in 19 Finnish families. Here we report the molecular background of CLD via characterization of five distinct mutations in the coding region of the lactase (LCT) gene. Twenty-seven patients out of 32 (84%) were homozygous for a nonsense mutation, c.4170T-->A (Y1390X), designated "Fin(major)." Four rare mutations--two that result in a predicted frameshift and early truncation at S1666fsX1722 and S218fsX224 and two point mutations that result in substitutions Q268H and G1363S of the 1,927-aa polypeptide--confirmed the lactase mutations as causative for CLD. These findings facilitate genetic testing in clinical practice and enable genetic counseling for this severe disease. Further, our data demonstrate that, in contrast to common adult-type hypolactasia (lactose intolerance) caused by a variant of the regulatory element, the severe infancy form represents the outcome of mutations affecting the structure of the protein inactivating the enzyme.

The allergenicity and antigenicity of cow milk proteins are age dependent. Because the nonspecific and specific factors inhibiting the passage of cow milk proteins through the epithelial layer of the intestine are deficient at birth, although developing during early infancy, allergy to cow milk may be acquired during the first year of life. Allergic reactivity to cow milk is lost during childhood in the vast majority of cases. This change may depend at least partly on the development of the local immune system of the gut producing antigen-specific IgA antibodies. Circulating IgG antibodies to cow milk proteins are always produced when an infant has cow milk in the diet but are not associated with allergy; their titer is reduced with age. Clinical challenge tests show that most cow milk-allergic patients react to several protein fractions of cow milk. A patient may have IgE antibodies to several fractions of cow milk, measured either by skin testing or by radioallergosorbent test. Likewise, various tests for cell-mediated immunity may show positive reactions to several fractions. No single major allergen is apparent in cow milk, according to either the challenge tests or laboratory procedures: casein, alpha-lactalbumin, and beta-lactoglobulin all show a high proportion of positive reactions.

Food antigens and enteroviruses are possible triggers of type 1 diabetes. Because permeability of the intestinal epithelium may facilitate contact of these antigens with the mucosal immune system, we set out to study intestinal permeability in patients with type 1 diabetes. Children with type 1 diabetes (n = 26, mean age 12 years, mean duration of disease 4 years) and 24 healthy age-matched control children were given mannitol and lactulose orally, and their intestinal permeability was measured as a percentage of this dose recovered in urine. Patients with type 1 diabetes did not differ in their permeability to lactulose, nor was their lactulose/mannitol ratio any different from that of controls. However, patients with type 1 diabetes who had the HLA-DQB 1*02 allele and, therefore, a higher risk for celiac disease (CD) absorbed significantly more mannitol (mean + 95% CI): 17.7% (15.2-20.2) than did those negative for this allele: 12.3% (8.2-16.4), p = 0.04. Their lactulose permeability was also higher: 0.30 (0.16-0.44) and 0.09% (0-0.18), respectively, p = 0.02. Although the differences in permeability reach statistical significance, there was still much overlap between the two groups in terms of actual laboratory values. The higher permeability of patients with the HLA-DQB1*02 allele suggests that these patients may be more prone to develop abnormal immune responses to food antigens.

Coeliac disease in adolescents has been associated with an increased prevalence of depressive and disruptive behavioural disorders, particularly in the phase before diet treatment. We studied the possible effects of a gluten-free diet on psychiatric symptoms, on hormonal status (prolactin, thyroidal function) and on large neutral amino acid serum concentrations in adolescents with coeliac disease commencing a gluten-free diet.Nine adolescents with celiac disease, aged 12 to 16 years, were assessed using the semi-structured K-SADS-Present and Lifetime Diagnostic interview and several symptom scales. Seven of them were followed at 1 to 2, 3, and 6 months on a gluten-free diet.Adolescent coeliac disease patients with depression had significantly lower pre-diet tryptophan/ competing amino-acid (CAA) ratios and free tryptophan concentrations, and significantly higher biopsy morning prolactin levels compared to those without depression. A significant decrease in psychiatric symptoms was found at 3 months on a gluten-free diet compared to patients' baseline condition, coinciding with significantly decreased coeliac disease activity and prolactin levels and with a significant increase in serum concentrations of CAAs.Although our results of the amino acid analysis and prolactin levels in adolescents are only preliminary, they give support to previous findings on patients with coeliac disease, suggesting that serotonergic dysfunction due to impaired availability of tryptophan may play a role in vulnerability to depressive and behavioural disorders also among adolescents with untreated coeliac disease.

Using a case-control design we have studied whether antibodies to cow's milk proteins are risk determinants for childhood-onset Type 1 (insulin-dependent) diabetes mellitus independent of early exposure to cow's milk formula and islet cell antibodies. Sera from 116 recentonset diabetic children and 112 age- and sex- matched control children were analysed for cow's milk protein IgA, IgG and IgM antibodies, β-lactoglobulin IgA and IgM antibodies and islet cell antibodies. The titres were compared to questionnaire data on duration of breast-feeding and introduction of formula feeding. Most antibody levels tended to be increased among diabetic compared to control children. This was statistically significant for cow's milk protein IgA antibodies (p <0.001) and β-ltoglobulin IgA antibodies (p <0.01) as well as for islet cell antibody-positivity which was found among 92% of the diabetic and 3% of control children. The differences in cow's milk protein antibodies as well as β-lactoglobulin antibodies were more pronounced among children with an early onset of Type 1 diabetes. Breast-feeding duration was significantly inversely related to the log of β-Mactoglobulin IgG (r = −0.16, p = 0.04) and the log of cow's milk protein IgA antibodies (r = −0.17, p<0.001). A positive correlation was found between formula feeding and the logarithm of β-lactoglobulin IgG antibodies (r = 0.22, p = 0.01) and the log of cow's milk protein IgA antibodies (r = 0.16, p = 0.04). In a multiple logistic regression analysis it was found that IgA antibodies to β-lactoglobulin and cow's milk protein were significantly related to the risk of Type 1 diabetes independent of islet cell antibodies. When introducing formula feeding before the age of 4 months as a variable in the regression it was shown that islet cell antibodies and β-lactoglobulin IgA antibodies were still significantly and independently related to an increased risk of diabetes whereas cow's milk protein IgA antibodies did not add further to the regression. It is concluded that β-lactoglobulin IgA antibodies are significantly associated with an increased risk of diabetes at a young age independent of islet cell antibody-status and of an early weaning to cow's milk formula. In genetically susceptible children early exposure to β-actoglobulin might be one trigger in the autoimmune process leading to development of Type 1 diabetes.

ABSTRACT. During a nutritional study of 198 infants, seven became allergic to cow's milk. The seven infants showed acute cutaneous manifestations during cow's milk challenge tests in hospital and six had increased levels of IgE cow's milk‐specific antibodies. Neither in the development of the levels of immunoglobulins G, A and M, nor in that of the cow's milk‐specific antibodies of these isotypes did these seven patients differ from the remaining infants. Beta‐lactoglobulin content and levels of cow's milk‐, and beta‐lactoglobulin‐specific antibodies and of immunoglobulins A, G and M were measured in samples of colostrum and milk from the mothers of the seven infants with cow's milk allergy and from a comparison group (non‐atopic mothers of non‐atopic infants). The milk of the mothers whose infants became allergic to cow's milk contained less IgA through the lactation: 95% confidence intervals of the groups did not overlap. The difference was most marked in the colostrum. All other measurements were similar in the two groups. This suggests that an infant is more likely to develop cow's milk allergy if the mother's colostrum had a low total IgA content.

Specific defense factors in breast milk together with length of breast-feeding and genetic predisposition may modulate the development of allergy. We studied whether IgA, soluble CD14 (sCD14), or transforming growth factor (TGF)-β in colostrum could affect the development of atopy in children up to age 4. From a cohort of 4676, we selected four groups of children with either long or short exclusive breast-feeding (>3.5 or <0.5 mo); these groups further differed in the presence or absence of atopic heredity. In colostrum from mothers, we measured total IgA, IgA antibodies to cow's milk (CM) and casein, sCD14, and TGF-β1 and -β2. The children were divided into three groups: those with no atopic symptoms or IgE, those with allergic symptoms, and those with both outcomes. Mothers of infants later showing atopic symptoms or, in addition, having IgE sensitization (verified atopy) had a lower concentration of IgA casein antibodies in their colostrum than did mothers of infants with no indication of atopy at age 4. Low concentration of IgA casein antibodies was a significant risk for verified atopy. sCD14 levels were lower in colostrum of mothers with infants developing atopic symptoms and IgE sensitization than of those of infants with no atopy. Specific IgA antibodies to CM antigens and sCD14 in colostrum significantly associated with the appearance of both symptomatic and verified atopy by age 4.

Abstract Background Human gastrointestinal mucosa regenerates vigorously throughout life, but the factors controlling cell fate in mature mucosa are poorly understood. GATA transcription factors direct cell proliferation and differentiation in many organs, and are implicated in tumorigenesis. GATA-4 and GATA-6 are considered crucial for the formation of murine gastrointestinal mucosa, but their role in human gastrointestinal tract remains unexplored. We studied in detail the expression patterns of these two GATA factors and a GATA-6 down-stream target, Indian hedgehog (Ihh), in normal human gastrointestinal mucosa. Since these factors are considered important for proliferation and differentiation, we also explored the possible alterations in their expression in gastrointestinal neoplasias. The expression of the carcinogenesis-related protein Indian hedgehog was also investigated in comparison to GATA factors. Methods Samples of normal and neoplastic gastrointestinal tract from children and adults were subjected to RNA in situ hybridization with 33 P labelled probes and immunohistochemistry, using an avidin-biotin immunoperoxidase system. The pathological tissues examined included samples of chronic and atrophic gastritis as well as adenomas and adenocarcinomas of the colon and rectum. Results GATA-4 was abundant in the differentiated epithelial cells of the proximal parts of the gastrointestinal tract but was absent from the distal parts. In contrast, GATA-6 was expressed throughout the gastrointestinal epithelium, and in the distal gut its expression was most intense at the bottom of the crypts, i.e. cells with proliferative capacity. Both factors were also present in Barrett's esophagus and metaplasia of the stomach. GATA-6 expression was reduced in colon carcinoma. Ihh expression overlapped with that of GATA-6 especially in benign gastrointestinal neoplasias. Conclusion The results suggest differential but overlapping functions for GATA-4 and GATA-6 in the normal gastrointestinal mucosa. Furthermore, GATA-4, GATA-6 and Ihh expression is altered in premalignant dysplastic lesions and reduced in overt cancer.

We followed 183 infants for two years, 31 of whom were breast fed less than three and a half months (median 70 days; short breast feeding group) and a further 31 of whom were exclusively breast fed for more than nine months (long breast feeding group). We assessed heredity for atopy, number of infections, and duration of breast feeding as determinants of atopy. During the first year of life 14 infants has signs of atopy. During the second year parents reported signs of atopy in a further 31. Heredity was the only significant predictor of atopy. Atopy was seen in 33% of infants with a positive heredity and in 16% without family history for atopy. The duration of breast feeding affected the incidence of atopy only among the infants without family history for atopy: fewer in the short breast feeding group (1/18) had atopy than in the long breast feeding group (5/13). Duration of breast feeding did not associate with incidence of respiratory infections. Diarrhoea was more common in the short breast feeding group than in the long breast feeding group during the first year of life. We conclude that prolonging exclusive breast feeding from the median of 70 days to nine months did not contribute to the prevention of infantile atopy and respiratory tract infections.

ABSTRACT: We followed 183 infants from birth to 2.3 yr of age. Of these infants 28 had recurrent otitis media (ROM), defined as five or more separate episodes of otitis media (OM) during the first 2 yr of life or four such episodes during their 2nd yr. The OM presented during their 1st yr (early-onset ROM) in 12 infants and during their 2nd yr (2nd yr ROM) in 16. Eighty infants had no OM and served as a comparison group. Regarding type of feeding, the infants with early-onset ROM did not differ from their age-matched pairs in the comparison group either 1 month before the first OM or at the time of first episode of OM. Exclusive breast-feeding did not prevent OM and early weaning was not a risk factor for ROM. Atopy was associated with ROM with a relative risk of 1.9 (95% confidence limits 1.2–3.2). It was particularly prevalent among the infants with early-onset ROM, in 67 versus in 25% in the comparison group (p < 0.01). During, the 2nd yr daily contact with five or more children was associated with ROM with a relative risk of 2.1 (1.3–3.3). The infants with 2nd-yr ROM were in daily contact with more children than the comparison group (mean 11 versus 5; p < 0.001). Parental smoking was more frequent among the infants with ROM than in the comparison group (54 versus 33%; p < 0.05). In the infants with early-onset ROM plasma concentration of IgM antibodies to cow's milk was highest at the age of 9 months, and the concentration of IgE was highest at the ages of 9 and 12 months. In conclusion atopy, not the type of feeding, is a risk factor for early-onset ROM, and daycare outside the home for ROM during the 2nd yr.

&lt;b&gt;&lt;i&gt;Background:&lt;/i&gt;&lt;/b&gt; Whether breast milk (BM) can protect against allergy has been studied extensively, with conflicting results. Variations in mothers’ BM composition may explain some of the conflicting results. Our aim was to assess the impact of maternal allergy and probiotic intervention on BM food antibodies, transforming growth factor (TGF)-β&lt;sub&gt;2&lt;/sub&gt; and interleukin (IL)-10 and their impact on allergy development in children until the ages of 2 and 5. &lt;b&gt;&lt;i&gt;Methods:&lt;/i&gt;&lt;/b&gt; We measured total IgA, IgA antibodies to cow’s milk (CM), casein, β-lactoglobulin and ovalbumin (OVA), TGF-β&lt;sub&gt;2&lt;/sub&gt; and IL-10 in 364 colostrum samples and 321 BM samples taken at 3 months from mothers participating in a prospective study evaluating the allergy-preventive effect of probiotics in a cohort with an increased risk for allergy. &lt;b&gt;&lt;i&gt;Results:&lt;/i&gt;&lt;/b&gt; CM, casein and OVA antibodies, TGF-β&lt;sub&gt;2&lt;/sub&gt; and IL-10 were detectable in most samples. Maternal allergy was associated with raised levels of IgA to casein (p = 0.04) and lower levels of TGF-β&lt;sub&gt;2&lt;/sub&gt; (p = 0.006) in mature BM. Probiotic supplementation was associated with increased IL-10 (p = 0.046) and decreased casein IgA antibodies (p = 0.027) in mature BM. High OVA IgA antibodies in colostrum were associated with the development of atopy by the age of 2, while low levels in mature BM were a significant risk factor for the development of eczema by the age of 2. TGF-β&lt;sub&gt;2&lt;/sub&gt; levels in BM constituted a risk for development of allergy by the age of 2. &lt;b&gt;&lt;i&gt;Conclusions:&lt;/i&gt;&lt;/b&gt; The immunologic composition of BM was only slightly affected by maternal atopy and could be altered by probiotic supplementation. Small effects of BM components on allergy development in children were evident.

The numbers of immunoglobulin-containing cells in jejunal biopsy specimens of 19 children with active coeliac disease aged 05 to 16.5 years were studied by direct immunofluorescence.Intestinal juice immunoglobulins were measured in 14 of these patients.The number of IgA-containing cells was twice and the number of IgM-containing cells 2.5 times that of age-matched controls.There were also more IgG-, IgE-, and IgD-containing cells in the jejunal mucosa of the coeliac patients, but the absolute numbers of these cells were low.The immunoglobulin content of the intestinal juice was not altered in coeliacs.A follow-up biopsy specimen was available from seven patients kept on a strict gluten-free diet for one to four months.A significant fall in the numbers of immunoglobulin-containing cells was seen, and they did not differ at that time from the controls.Two patients were followed until full normalization of the jejunal structure and they had normal numbers of immunoglobulin-containing cells.In children with coeliac disease in contrast to adult coeliacs, the study shows that the IgA-producing system is quantitatively stimulated during gluten challenge.The rapid drop in the numbers of immunoglobulin-containing cells after gluten withdrawal suggests that there is no quantitative abnormality in the local immunoglobulin-producing system of the gut in coeliac disease.Recently, interest has been focused on the local immunoglobulin-producing system of the gut.The distribution of immunoglobulin-containing cells in the mucosa of the gastrointestinal tract of normal persons is well established (Crabbe, Carbonara, and Heremans, 1965; Crabbe and Heremans, 1966), and the changes taking place in this system have been investigated from early infancy (Crabbe, 1967; Savilahti, 1972a).These cells have also been studied in various gastrointestinal diseases (Odgers and Wangel, 1968; Soltoft, 1969), especially in coeliac disease.Rubin, Fauci, Sleisenger, and Jefferies (1965) did not observe any difference between immunoglobulincontaining cells of adult coeliacs and controls.Excess of IgM-containing cells was noted by Soltoft (1970), Douglas, Crabbe, and Hobbs (1970), and Pettingale (1971), and in the two latter studies reduction in the numbers of IgA-containing cells was also found in the jejunal mucosa of adult coeliac patients.In contrast, in children with coeliac

The first few months of life represent a window of opportunity for neonatal oral tolerance induction. During this critical time, breast milk is a rich source of immunomodulatory factors, including TGF-β, which has been shown to influence the maturation of the mucosal immune system of the neonates.1 Reports on breast milk cytokines, chemokines, and growth factors in cow's milk allergy (CMA), including both IgE- and non-IgE-mediated mechanisms, are largely lacking, although IL-5 and IL-13 were identified as risk factors for asthma and eotaxin for atopic dermatitis.

Objectives: Several studies have reported that the intestinal microbiota composition of celiac disease (CD) patients differs from healthy individuals. The possible role of gut microbiota in the pathogenesis of the disease is, however, not known. Here, we aimed to assess the possible differences in early fecal microbiota composition between children that later developed CD and healthy controls matched for age, sex and HLA risk genotype.Materials and methods: We used 16S rRNA gene sequencing to examine the fecal microbiota of 27 children with high genetic risk of developing CD. Nine of these children developed the disease by the age of 4 years. Stool samples were collected at the age of 9 and 12 months, before any of the children had developed CD. The fecal microbiota composition of children who later developed the disease was compared with the microbiota of the children who did not have CD or associated autoantibodies at the age of 4 years. Delivery mode, early nutrition, and use of antibiotics were taken into account in the analyses.Results: No statistically significant differences in the fecal microbiota composition were found between children who later developed CD (n = 9) and the control children without disease or associated autoantibodies (n = 18).Conclusions: Based on our results, the fecal microbiota composition at the age of 9 and 12 months is not associated with the development of CD. Our results, however, do not exclude the possibility of duodenal microbiota changes or a later microbiota-related trigger for the disease.

This study assesses the differences in composition of human milk oligosaccharides of mothers who received either probiotic supplements or placebos in the late stages of pregnancy.

Breast milk contains immune factors that compensate for the underdeveloped defenses of the gut of the newborn infant.We sought to study the importance of these factors in the immune responses of infants with cows' milk allergy (CMA) to the proteins in cows' milk (CM).We prospectively followed the development of CMA in 6209 healthy infants and collected samples of colostrum from mothers. Samples from mothers of infants with CMA and from control subjects were analyzed for immunoglobulins, CM-specific antibodies, and cytokines. In infants with CMA, correlations between the concentration of transforming growth factor (TGF)-beta1 in colostrum and the extent of the immune response to CM proteins were studied.The concentration of TGF-beta1 in colostrum samples from mothers of infants with IgE-mediated CMA (n = 65) was lower (mean, 589 pg/mL; 95% confidence interval [CI], 413-840) than from mothers of infants with non-IgE-mediated CMA (n = 37; mean, 1162 pg/mL; 95% CI, 881-1531; t = 2.57, P =.012). In 126 control subjects the mean concentration was 807 pg/mL (95% CI, 677-963). In the infants with CMA (n = 96-100), the concentration of TGF-beta1 in colostrum was positively correlated with IgA antibodies to beta-lactoglobulin and IgG antibodies to alpha-casein and whole formula and negatively with the diameter of a skin prick test response to CM and lymphocyte stimulation indices to alpha-casein and beta-lactoglobulin.In an infant prone to having CMA, the TGF-beta1 content of mother's colostrum may promote IgG-IgA antibody production and inhibit IgE- and cell-mediated reactions to CM.

Early feeding with cows’ milk (CM) may cause cows’ milk allergy (CMA). Breast milk contains many immune factors which compensate for the undeveloped defence mechanisms of the gut of the newborn infant. We studied the effect of supplementary CM feeding at the maternity hospital on the subsequent incidence of CMA, the effects of formula and breast feeding on the subsequent immunologic types of CMA, and the importance of immune factors present in colostrum in the immune responses of infants with CMA. In a cohort of 6209 infants, 824 were exclusively breast-fed and 87% required supplementary milk while in the maternity hospital: 1789 received CM formula, 1859 pasteurized human milk, and 1737 whey hydrolysate formula. The cumulative incidence of CMA, verified by a CM elimination-challenge test, was 2.4% in the CM, 1.7% in the pasteurized human milk and 1.5% in the whey hydrolysate group. Among these infants, exposure to CM at hospital and a positive atopic heredity increased the risk of CMA. Of the exclusively breast-fed infants, 2.1 % had CMA. Risk factors for the development of IgE-mediated CMA were:exposure to CM at hospital, breast-feeding during the first 8 weeks at home either exclusively or combined with infrequent exposure to small amounts of CM and long breast-feeding. The content of transforming growth factor- β 1(TGF-β1) in colostrum from mothers of infants with IgE-mediated CMA was lower than from mothers of infants with non-IgE-mediated CMA. In infants with CMA, TGF- β 1 in colostrum negatively correlated with the result of skin prick test and the stimulation of peripheral blood mononuclear cells to CM, but positively with infants’ IgA and IgG antibodies to CM proteins. Feeding of CM formula at maternity hospital increases the risk of CMA, but exclusive breast-feeding does not eliminate the risk. Prolonged breast-feeding exclusively or combined with infrequent exposure to small amounts of CM during the first 8 weeks induces the development of IgE-mediated CMA. Colostral TGF- β 1 may inhibit IgE-and cell mediated reactions and promote IgG-IgA antibody production to CM in infants prone to developing CMA.

SummaryCongenital lactase deficiency (CLD) is an autosomal recessive, gastrointestinal disorder characterized by watery diarrhea starting during the first 1–10 d of life, in infants fed lactose-containing milks. Since 1966, 42 patients have been diagnosed in Finland. CLD is the most severe form of lactase deficiency, with an almost total lack of lactase-phlorizin hydrolase (LPH) activity on jejunal biopsy. In adult-type hypolactasia, the most common genetic enzyme deficiency in humans, this enzyme activity is reduced to 5%–10%. Although the activity of intestinal LPH has been found to be greatly reduced in both forms, the molecular pathogenesis of lactase deficiencies is unknown. On the basis of the initial candidate-gene approach, we assigned the CLD locus to an 8-cM interval on chromosome 2q21 in 19 Finnish families. At the closest marker locus, a specific allele 2 was present in 92% of disease alleles. On the basis of a genealogical study, the CLD mutation was found to be enriched in sparsely populated eastern and northern Finland, because of a founder effect. The results of both the genealogical study and the haplotype analysis indicate that one major mutation in a novel gene causes CLD in the Finnish population. Consequently, the critical region could be restricted further, to an ∼350-kb interval, by ancient-haplotype and linkage-disequilibrium analyses. Surprisingly, the LPH gene was shown to lie outside the critical CLD region, excluding it as a causative gene for CLD. The LPH locus was found to reside >2 Mb from the critical CLD region. Congenital lactase deficiency (CLD) is an autosomal recessive, gastrointestinal disorder characterized by watery diarrhea starting during the first 1–10 d of life, in infants fed lactose-containing milks. Since 1966, 42 patients have been diagnosed in Finland. CLD is the most severe form of lactase deficiency, with an almost total lack of lactase-phlorizin hydrolase (LPH) activity on jejunal biopsy. In adult-type hypolactasia, the most common genetic enzyme deficiency in humans, this enzyme activity is reduced to 5%–10%. Although the activity of intestinal LPH has been found to be greatly reduced in both forms, the molecular pathogenesis of lactase deficiencies is unknown. On the basis of the initial candidate-gene approach, we assigned the CLD locus to an 8-cM interval on chromosome 2q21 in 19 Finnish families. At the closest marker locus, a specific allele 2 was present in 92% of disease alleles. On the basis of a genealogical study, the CLD mutation was found to be enriched in sparsely populated eastern and northern Finland, because of a founder effect. The results of both the genealogical study and the haplotype analysis indicate that one major mutation in a novel gene causes CLD in the Finnish population. Consequently, the critical region could be restricted further, to an ∼350-kb interval, by ancient-haplotype and linkage-disequilibrium analyses. Surprisingly, the LPH gene was shown to lie outside the critical CLD region, excluding it as a causative gene for CLD. The LPH locus was found to reside >2 Mb from the critical CLD region.

Elevated levels of antibodies to cow's milk proteins, i.e., β-lactoglobulin (BLG) and bovine serum albumin (BSA), have been associated with IDDM. We observed enhanced cellular immune response by a proliferation test of peripheral blood mononuclear cells to BLG in 22 of 40 (55%) patients with newly diagnosed IDDM compared with 7 of 32 healthy children (22%) (P = 0.004, X2 test). The median stimulation index to BLG was 3.3 in patients and 1.5 in healthy children (P = 0.003, Mann-Whitney U test). No difference was found in cellular reactivity to other cow's milk proteins, such as BSA or α-casein, or to a dietary immunogenic protein, ovalbumin. Cellular responsiveness to BLG was not associated with HLA-DQB1* risk alleles of IDDM, which suggests that immune response to the protein does not only reflect the accumulation of these HLA alleles in the patients with IDDM. We suggest that enhanced cellular immune response to dietary BLG may reflect a disturbance in the regulation of immune response to oral antigens in IDDM. This kind of defect may play a fundamental role in the development of β-cell autoimmunity in IDDM.

Migration of lymphocytes to the pancreas is a prerequisite for insulitis in IDDM. Mucosal vascular addressin (MAdCAM-1), involved in the recirculation of lymphocytes to the gut, has been found in the inflamed islets in NOD mice. In humans, triggers of the gut immune system (e.g., early exposure to cow's milk proteins in infancy, exposure to enteroviral infections) have been associated with IDDM. To study the possible link between the gut immune system and IDDM, we tested the expression of the α4β7-integrin, a homing receptor for MAdCAM-1, on GAD65-reactive lymphocytes. Using immunomagnetic cell sorting, we depleted the lymphocytes with high expression of α4β7-integrin in the peripheral blood mononuclear cell population from IDDM patients and patients with autoimmune polyendocrine disease type 1 (APD-I). The depletion led to a marked decrease (mean 70%) in the cellular response against GAD65 in three of six IDDM patients and in one subject at high risk for IDDM. A decrease of 37% in the GAD response was observed after depletion in the case of one APD-I patient who also had IDDM. Cellular response to tetanus toxoid increased in the majority of patients as well as in three control subjects studied. We demonstrated that a remarkable population of islet cell antigen–reactive lymphocytes express the gut-specific homing receptor, which emphasizes the role of gut immunity in IDDM. The manipulation of the gut immune system is therefore proposed as a tool for modulation of the autoimmunity against pancreatic β-cells in IDDM.

Background: The long‐term effect of early feeding on atopic sensitization is still unsolved. The aim of this study was to evaluate the long‐term effect of breastfeeding on atopy in groups of 4‐year‐old children stratified by atopic heredity. Methods: We collected four groups of 4‐year‐old children from a birth cohort: two groups with differing backgrounds of atopic heredity, all exclusively breast‐fed for at least 3 months; and two groups with differing atopic heredity, but all fed with cow's milk‐based formula during their first weeks. The data were collected with a questionnaire, skin prick testing, and measurement of serum total and allergen‐specific IgE levels. Results: Breastfeeding significantly decreased the risk of allergic rhino‐conjunctivitis [odds ratio (OR) 0.41, 95% confidence interval (CI) 0.18–0.95] and sensitization to furred pets, as measured by skin prick results, in children with atopic heredity, whereas in children without atopic heredity, breastfeeding was related to an increased risk of symptomatic atopy (OR 2.57, 95% CI 1.16–5.70), and high serum IgE values. A significant interaction was found between heredity and breastfeeding. Conclusions: The long‐term effect of breastfeeding was dual: in children with atopic heredity, breastfeeding protected against atopy, whereas in children without atopic heredity, it increased the risk of atopy.

Diabetes/Metabolism ReviewsVolume 9, Issue 4 p. 269-278 Article The case for elimination of cow's milk in early infancy in the prevention of type 1 diabetes: The finnish experience H. K. Åkerblom, Corresponding Author H. K. Åkerblom The Children's Hospital, II Department of Pediatrics, University of Helsinki, FinlandThe Children's Hospital, II Department of Pediatrics, University of Helsinki, Stenbäckinkatu 11, 00290 Helsinki, FinlandSearch for more papers by this authorT. T. Saukkonen, T. T. Saukkonen The Children's Hospital, II Department of Pediatrics, University of Helsinki, Finland The Children's Hospital, I Department of Pediatrics, University of Helsinki, FinlandSearch for more papers by this authorO. Vaarala, O. Vaarala The Children's Hospital, II Department of Pediatrics, University of Helsinki, Finland The Children's Hospital, I Department of Pediatrics, University of Helsinki, FinlandSearch for more papers by this authorE. Savilahti, E. Savilahti The Children's Hospital, I Department of Pediatrics, University of Helsinki, FinlandSearch for more papers by this authorA. Paganus, A. Paganus The Children's Hospital, I Department of Pediatrics, University of Helsinki, FinlandSearch for more papers by this authorS. M. Virtanen, S. M. Virtanen The Children's Hospital, II Department of Pediatrics, University of Helsinki, Finland The Children's Hospital, Division of Nutrition, Department of Applied Chemistry and Microbiology, University of Helsinki, FinlandSearch for more papers by this authorK. Teramo, K. Teramo The Children's Hospital, I and II Departments of Obstetrics and Gynecology, University of Helsinki, FinlandSearch for more papers by this authorM. Knip, M. Knip The Children's Hospital, Department of Pediatrics, University of Oulu, FinlandSearch for more papers by this authorJ. Karjalainen, J. Karjalainen The Children's Hospital, Department of Pediatrics, University of Oulu, FinlandSearch for more papers by this authorJ. Ilonen, J. Ilonen The Children's Hospital, Department of Virology, University of Turku, FinlandSearch for more papers by this authorH. Reijonen, H. Reijonen The Children's Hospital, Department of Virology, University of Turku, FinlandSearch for more papers by this authorA. Reunanen, A. Reunanen The Children's Hospital, The Social Insurance Institution, Research and Development Unit, Helsinki, FinlandSearch for more papers by this author H. K. Åkerblom, Corresponding Author H. K. Åkerblom The Children's Hospital, II Department of Pediatrics, University of Helsinki, FinlandThe Children's Hospital, II Department of Pediatrics, University of Helsinki, Stenbäckinkatu 11, 00290 Helsinki, FinlandSearch for more papers by this authorT. T. Saukkonen, T. T. Saukkonen The Children's Hospital, II Department of Pediatrics, University of Helsinki, Finland The Children's Hospital, I Department of Pediatrics, University of Helsinki, FinlandSearch for more papers by this authorO. Vaarala, O. Vaarala The Children's Hospital, II Department of Pediatrics, University of Helsinki, Finland The Children's Hospital, I Department of Pediatrics, University of Helsinki, FinlandSearch for more papers by this authorE. Savilahti, E. Savilahti The Children's Hospital, I Department of Pediatrics, University of Helsinki, FinlandSearch for more papers by this authorA. Paganus, A. Paganus The Children's Hospital, I Department of Pediatrics, University of Helsinki, FinlandSearch for more papers by this authorS. M. Virtanen, S. M. Virtanen The Children's Hospital, II Department of Pediatrics, University of Helsinki, Finland The Children's Hospital, Division of Nutrition, Department of Applied Chemistry and Microbiology, University of Helsinki, FinlandSearch for more papers by this authorK. Teramo, K. Teramo The Children's Hospital, I and II Departments of Obstetrics and Gynecology, University of Helsinki, FinlandSearch for more papers by this authorM. Knip, M. Knip The Children's Hospital, Department of Pediatrics, University of Oulu, FinlandSearch for more papers by this authorJ. Karjalainen, J. Karjalainen The Children's Hospital, Department of Pediatrics, University of Oulu, FinlandSearch for more papers by this authorJ. Ilonen, J. Ilonen The Children's Hospital, Department of Virology, University of Turku, FinlandSearch for more papers by this authorH. Reijonen, H. Reijonen The Children's Hospital, Department of Virology, University of Turku, FinlandSearch for more papers by this authorA. Reunanen, A. Reunanen The Children's Hospital, The Social Insurance Institution, Research and Development Unit, Helsinki, FinlandSearch for more papers by this author First published: December 1993 https://doi.org/10.1002/dmr.5610090407Citations: 47AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onEmailFacebookTwitterLinkedInRedditWechat References 1 Eisenbarth GS: Type 1 diabetes mellitus: a chronic autoimmune disease. 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A high prevalence of depressive symptoms, hypothetically related to serotonergic dysfunction, has been reported among adults with celiac disease. The authors used semistructured psychiatric interviews and symptom measurement scales to study mental disorders in 29 adolescents with celiac disease and 29 matched comparison subjects. Relative to the comparison subjects, the celiac disease patients had significantly higher lifetime prevalences of major depressive disorder (31% versus 7%) and disruptive behavior disorders (28% versus 3%). In most cases these disorders preceded the diagnosis of celiac disease and its treatment with a gluten-free diet. The prevalence of current mental disorders was similar in both groups. Celiac disease in adolescents is associated with an increased prevalence of depressive and disruptive behavioral disorders, particularly in the phase before diet treatment.

In a double blind crossover study 10 children with infantile colic were fed breast milk and cow's milk formula, untreated and treated with lactase. Colic was present on 71% of breast milk and 89% of cow's milk days. Daily duration and severity of colic did not differ for milk preparations.

A total of 16 children with Shwachman's syndrome were studied over a period of 17 years. Eight cases were detected in autopsy at 6 to 15 months; all had died of cardiac failure due to myocardial lesions. The left ventricles showed necrosis of myofibres in large areas and the pancreas was atrophic and replaced by adipose tissue. The other eight patients included two siblings of deceased cases. Only one of these showed transient cardiac failure and no significant nutritional deficiencies were found. They had steatorrhoea due to failure of the exocrine pancreas and either constant (6 cases) or cyclic (2 cases) neutropenia. The steatorrhoea improved with age. Pyogenic infections, mainly otitis media were frequent during the first three years of life. Measurements of humoral and cell-mediated immunity were normal, but in addition to low numbers of neutrophils, the neutrophilic chemotaxis was depressed in all seven patients tested. Skin lesions, hepatic inflammation, and growth tended to improve with age. The family data of the patients is consistent with an autosomal recessive trait inheritance.

To correlate the C/T(-13910) variant, associated with lactase persistence/non-persistence (adult-type hypolactasia) trait, with intestinal disaccharidase activities in different age groups of the adult population.Intestinal biopsies were obtained from 222 adults aged 18 to 83 years undergoing upper gastrointestinal endoscopy because of unspecified abdominal complaints. The biopsies were assayed for lactase, sucrase and maltase activities and genotyped for the C/T(-13910) variant using PCR-minisequencing.There was a significant correlation between lactase activity and the C/T(-13910) variant (P < 0.00001). The mean level of lactase activity among subjects with C/C(-13910) genotype was 6.86 +/- 0.35 U/g, with C/T(-13910) genotype 37.8 +/- 1.4 U/g, and with T/T(-13910) genotype 57.6 +/- 2.4 U/g protein, showing a trimodal distribution of this enzyme activity. Significant differences were also observed in maltase activities among individuals with different C/T(-13910) genotypes (P = 0.005). In contrast, in sucrase activity, no significant differences emerged between the C/T(-13910) genotypes (P = 0.14). There were no statistical differences in lactase (P = 0.84), sucrase (P = 0.18), or maltase activity (P = 0.24) among different age groups. In the majority (> 84%) of the patients with the C/C(-13910) genotype associated with lactase non-persistence, the lactase activity was less than 10 U/g protein.Our study demonstrates a statistically significant correlation between the C/T(-13910) genotype and lactase activity and this correlation is not affected by age in adults but the cut-off value of 20 U/g protein used for the diagnosis of lactase non-persistence might be too high.

Abstract Background Both the initiation and maintenance of breastfeeding have been reported to be negatively affected by maternal type 1 diabetes (T1D). The aim of this study was to prospectively examine the breastfeeding patterns among mothers with and without T1D participating in a large international randomized infant feeding trial (TRIGR). Methods Families with a member affected by T1D and with a newborn infant were invited into the study. Those who had HLA‐conferred genetic susceptibility for T1D tested at birth with gestation &gt; 35 weeks and were healthy were eligible to continue in the trial. Among the 2160 participating children, 1096 were born to women with T1D and 1064 to unaffected women. Information on infant feeding was acquired from the family by frequent prospective dietary interviews. Results Most (&gt;90%) of the infants of mothers with and without T1D were initially breastfed. Breastfeeding rates declined more steeply among mothers with than without T1D being 50 and 72% at 6 months, respectively. Mothers with T1D were younger, less educated and delivered earlier and more often by caesarean section than other mothers ( p &lt; 0.01). After adjusting for all these factors associated with the termination of breastfeeding, there was no difference in the duration of breastfeeding among mothers with and without T1D. Conclusions Maternal diabetes status per se was not associated with shorter breastfeeding. The lower duration of breastfeeding in mothers with T1D is largely explained by their more frequent caesarean sections, earlier delivery and lower age and education. Copyright © 2010 John Wiley &amp; Sons, Ltd.

ABSTRACT. Verkasalo, M., Kuitunen, P., Savilahti, E. and Tiilikainen, A. (Children's Hospital, University of Helsinki, Finland). Changing pattern of cow's milk intolerance. Acta Paediatr Scand, 70: 289, 1981.–The rapid changeover to commercial adapted infant formulae which took place in Finland between 1973 and 1975 was studied as a factor in the occurrence of severe intestinal cow's milk intolerance (CMI). Of infants treated for CMI in 1962‐73, ninety‐three percent (25/27) were on homemade or unadapted formulae. The admission rate for CMI in these years was 0.22/1 000 liveborn infants breast fed less than six months. During 1974‐77 the corresponding figure was 0.56, with 85 % of the patients (18/26) on adapted cow's milk formulae. The patients treated before 1974 had a longer symptomatic period before admission, greater growth retardation and more severe intestinal damage than those seen during and after 1974. This is believed to reflect mainly the increasing awareness of CMI on the part of both laymen and the medical profession. In the history of 2/3 of the patients at least one of the following conditions was noted: non‐breast feeding, infectious gastroenteritis, praematurity, 21‐trisomy, prior intra‐abdominal surgery, Hirschsprung's disease, and atopic disease in family members. The long follow‐up averaging over four years revealed four patients with coeliac disease. In one of these the proximal jejunal mucosa was normal after two years on gluten‐containing diet, but he showed a mucosal relapse as late as between 2 to 4 years on normal diet.

To cite this article: Kukkonen AK, Kuitunen M, Savilahti E, Pelkonen A, Malmberg P, Mäkelä M. Airway inflammation in probiotic‐treated children at 5 years. Pediatr Allergy and Immunol 2011; 22 : 249–251. Early treatment of new‐born high‐risk children with certain probiotic strains has reduced the risk of atopic eczema. Whether probiotics reduce risk for airway inflammation in long term is not known. We aimed at studying the effect of probiotic treatment during the six first months of life on airway inflammation at age 5 yr. In a randomized double‐blind allergy prevention trial between 2000 and 2007 in Helsinki, Finland, we gave a probiotic combination, plus pre‐biotics, or placebo, to 1018 children during 6 months from birth. At age 5, we measured exhaled nitric oxide (FE NO ) in a randomized sub‐population of 160 children. Allergic diseases and IgE‐sensitization were assessed in all infants. FE NO did not differ between probiotic and placebo groups, median (interquartile range, IQR) 5.45 (4.3–7.3) vs. 5.70 (3.9–6.8) ppb, p = 0.22. FE NO was elevated among those suffering from asthma during the first 5 yr than in healthy non‐sensitized children (p = 0.009). FE NO correlated positively with serum total and allergen‐specific IgE concentrations. Early intervention with probiotics and pre‐biotics does not affect airway inflammation later in childhood.

Abstract Cow’s milk allergy (CMA) affects 2–3% of infants. It resolves in the great majority spontaneously during childhood. CMA encompasses a spectrum of clinical and immunologic characteristics. Non‐IgE‐mediated allergy typically resolves earlier than IgE‐mediated allergy. The most documented prognostic characteristic is that intense‐specific IgE response predicts persistence of CMA. Low serum levels of cow’s milk (CM)‐specific IgG4 are also associated with persistent CMA. Natural development of tolerance involves an immunologic shift where Th2 responses diminish, and Th1 as well as T regulatory cell responses strengthen. Accordingly, specific IgE levels decrease and specific IgG4, possibly also IgA, levels increase in serum. Specific oral immunotherapy (OIT) with CM induces desensitization in most cases where spontaneous recovery has not yet occurred. Data on long‐term tolerance induction are still scarce. According to current research data, the immunologic changes induced by OIT resemble those seen during natural development of tolerance.