AIMS/HYPOTHESIS: We previously detected indications that beta cell function is protected by gluten-free diet (GFD) introduced shortly after the onset of childhood type 1 diabetes. The present aim was to assess whether GFD was associated with changes in the gut bacteriome composition and in its functional capacity, and whether such changes mediated the observed effects of GFD on beta cell function. METHODS: Forty-five children (aged 10.2 ± 3.3 years) were recruited into a self-selected intervention trial primarily focused on determining the role of GFD on beta cell preservation ( ClinicalTrials.gov NCT02867436). Stool samples were collected prior to the dietary intervention and then at 3-month intervals. A total of 128 samples from the GFD group and 112 from the control group were analysed for bacteriome 16S rDNA community profiles, the bacteriome functional capacity was predicted using PICRUSt2 and actual gut metabolome profiles measured using NMR. Intestinal permeability was assessed using serum zonulin concentrations at 1, 6 and 12 months and lactulose/mannitol tests at the end of intervention. Dietary questionnaires were used to ensure that the dietary intervention did not result in differences in energy or nutrient intake. RESULTS: The bacteriome community composition changed during the intervention with GFD: of abundant genera, a 3.3-fold decrease was noted for Bifidobacterium genus (adjusted p=1.4 × 10-4 in a DESeq2 model, p=0.026 in generalised estimating equations model), whereas a 2.4-fold increase was observed in Roseburia (adjusted p=0.02 in DESeq2 model, p=0.002 in generalised estimating equations model). The within-sample (alpha) diversity did not change, and there was no statistically significant clustering of GFD samples in the ordination graphs of beta diversity. Neither of the genera changes upon GFD intervention showed any association with the pace of beta cell loss (p>0.50), but of the remaining taxa, several genera of Bacteroidaceae family yielded suggestive signals. The faecal metabolome profile ordination correlated with that of bacteriomes but did not associate with GFD or categories of beta cell preservation. There was no indication of changes in gut permeability. CONCLUSIONS/INTERPRETATION: The bacteriome reacted to GFD, but the changes were unrelated to the pace of beta cell capacity loss. The previously observed moderately protective effect of GFD is therefore mediated through other pathways.

Department of Carbohydrates and Cereals University of Chemistry and Technology Prague Czechia

Department of Food Science Czech University of Life Sciences Prague Czechia

Department of Pediatrics 2nd Faculty of Medicine Charles University and Motol University Hospital Prague Czechia

Department of Pediatrics 3rd Faculty of Medicine Charles University and Kralovske Vinohrady University Hospital Prague Czechia

Department of Probability and Mathematical Statistics Faculty of Mathematics and Physics Charles University Prague Czechia

Zobrazit více v PubMed

Neuman V, Pruhova S, Kulich M et al (2020) Gluten-free diet in children with recent-onset type 1 diabetes: A 12-month intervention trial. Diabetes Obes Metab 22(5):866–872. https://doi.org/10.1111/dom.13974 DOI

Bonder MJ, Tigchelaar EF, Cai X et al (2016) The influence of a short-term gluten-free diet on the human gut microbiome. Genome Med 8(1):45. https://doi.org/10.1186/s13073-016-0295-y DOI

Lammers KM, Lu R, Brownley J et al (2008) Gliadin induces an increase in intestinal permeability and zonulin release by binding to the chemokine receptor CXCR3. Gastroenterology 135(1):194–204 e193. https://doi.org/10.1053/j.gastro.2008.03.023 DOI

El Asmar R, Panigrahi P, Bamford P et al (2002) Host-dependent zonulin secretion causes the impairment of the small intestine barrier function after bacterial exposure. Gastroenterology 123(5):1607–1615. https://doi.org/10.1053/gast.2002.36578 DOI

Greenbaum CJ, Mandrup-Poulsen T, McGee PF et al (2008) Mixed-meal tolerance test versus glucagon stimulation test for the assessment of beta-cell function in therapeutic trials in type 1 diabetes. Diabetes Care 31(10):1966–1971. https://doi.org/10.2337/dc07-2451 DOI

Kozich JJ, Westcott SL, Baxter NT, Highlander SK, Schloss PD (2013) Development of a dual-index sequencing strategy and curation pipeline for analyzing amplicon sequence data on the MiSeq Illumina sequencing platform. Appl Environ Microbiol 79(17):5112–5120. https://doi.org/10.1128/AEM.01043-13 DOI

Callahan BJ, McMurdie PJ, Rosen MJ, Han AW, Johnson AJ, Holmes SP (2016) DADA2: High-resolution sample inference from Illumina amplicon data. Nat Methods 13(7):581–583. https://doi.org/10.1038/nmeth.3869 DOI

Jaimes JD, Slavickova A, Hurych J et al (2021) Stool metabolome-microbiota evaluation among children and adolescents with obesity, overweight, and normal-weight using 1H NMR and 16S rRNA gene profiling. PLoS One 16(3):e0247378. https://doi.org/10.1371/journal.pone.0247378 DOI

Sequeira IR, Lentle RG, Kruger MC, Hurst RD (2014) Standardising the lactulose mannitol test of gut permeability to minimise error and promote comparability. PLoS One 9(6):e99256. https://doi.org/10.1371/journal.pone.0099256 DOI

Fasano A, Not T, Wang W et al (2000) Zonulin, a newly discovered modulator of intestinal permeability, and its expression in coeliac disease. Lancet 355(9214):1518–1519. https://doi.org/10.1016/S0140-6736(00)02169-3 DOI

Comino I, Fernandez-Banares F, Esteve M et al (2016) Fecal gluten peptides reveal limitations of serological tests and food questionnaires for monitoring gluten-free diet in celiac disease patients. Am J Gastroenterol 111(10):1456–1465. https://doi.org/10.1038/ajg.2016.439 DOI

Cuschieri S (2019) The STROBE guidelines. Saudi J Anaesth 13(Suppl 1):S31–S34. https://doi.org/10.4103/sja.SJA_543_18 DOI

De Palma G, Nadal I, Collado MC, Sanz Y (2009) Effects of a gluten-free diet on gut microbiota and immune function in healthy adult human subjects. Br J Nutr 102(8):1154–1160. https://doi.org/10.1017/S0007114509371767 DOI

Golfetto L, de Senna FD, Hermes J, Beserra BT, Franca Fda S, Martinello F (2014) Lower bifidobacteria counts in adult patients with celiac disease on a gluten-free diet. Arq Gastroenterol 51(2):139–143. https://doi.org/10.1590/s0004-28032014000200013 DOI

Di Cagno R, De Angelis M, De Pasquale I et al (2011) Duodenal and faecal microbiota of celiac children: molecular, phenotype and metabolome characterization. BMC Microbiol 11(1):219. https://doi.org/10.1186/1471-2180-11-219 DOI

Caminero A, Herran AR, Nistal E et al (2014) Diversity of the cultivable human gut microbiome involved in gluten metabolism: isolation of microorganisms with potential interest for coeliac disease. FEMS Microbiol Ecol 88(2):309–319. https://doi.org/10.1111/1574-6941.12295 DOI

Larretxi I, Churruca I, Navarro V et al (2020) Effect of analytically measured fiber and resistant starch from gluten-free products on the diets of individuals with celiac disease. Nutrition 70:110586. https://doi.org/10.1016/j.nut.2019.110586 DOI

Walker AW, Ince J, Duncan SH et al (2011) Dominant and diet-responsive groups of bacteria within the human colonic microbiota. ISME J 5(2):220–230. https://doi.org/10.1038/ismej.2010.118 DOI

Hansen LBS, Roager HM, Sondertoft NB et al (2018) A low-gluten diet induces changes in the intestinal microbiome of healthy Danish adults. Nat Commun 9(1):4630. https://doi.org/10.1038/s41467-018-07019-x DOI

Thorsen J, Brejnrod A, Mortensen M et al (2016) Large-scale benchmarking reveals false discoveries and count transformation sensitivity in 16S rRNA gene amplicon data analysis methods used in microbiome studies. Microbiome 4(1):62. https://doi.org/10.1186/s40168-016-0208-8 DOI

Ajamian M, Steer D, Rosella G, Gibson PR (2019) Serum zonulin as a marker of intestinal mucosal barrier function: May not be what it seems. PLoS One 14(1):e0210728. https://doi.org/10.1371/journal.pone.0210728 DOI

Tatucu-Babet OA, Forsyth A, Owen E et al (2020) Serum zonulin measured by enzyme-linked immunosorbent assay may not be a reliable marker of small intestinal permeability in healthy adults. Nutr Res 78:82–92. https://doi.org/10.1016/j.nutres.2020.05.003 DOI

Gluten-Free Diet Induces Small-Scale Changes Across Multiple T-Cell Subsets in NOD Mice

Zobrazit více v PubMed

ClinicalTrials.gov
NCT02867436