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. Author manuscript; available in PMC: 2024 Apr 12.
Published in final edited form as: Cell Host Microbe. 2023 Mar 29;31(4):554–570.e7. doi: 10.1016/j.chom.2023.03.006

Figure 3. Restriction of intestinal microbiome maturation stunts cellular and humoral immune system development.

Figure 3.

A. Representative flow cytometry plots of CD4+Foxp3+RORγ+Helios− peripheral Treg (pTreg) cell populations. Cells were gated on CD45+TCRβ+CD19−CD8−CD4+Foxp3+. B. Percentage of pTregs in the small intestine, cecum and large intestine lamina propria of germfree, PedsCom and CMCom mice during ontogeny. N = 3 for day 21 germfree cecum, N ≥ 4 per other communities and timepoints. Lines represent median value for each community. Mann-Whitney-Wilcoxon test *p<0.05, **p<0.01, ***p<0.001. C. Percentage of pTregs in the small intestine, cecum and large intestine lamina propria of germfree, CMCom, PedsCom and Oligo-MM12 7-12-week-old adult mice. N ≥ 4 for germfree tissues, N ≥ 6 per other communities and timepoints. D. Tumor necrosis factor alpha (TNFα) transcript levels in distal small intestinal tissue of germfree, PedsCom and CMCom mice during weaning. Relative quantitation (RQ) represents the fold change at day 21 and day 28 relative to mean baseline transcript levels at day 14 in each community. Day 21 represents pooled data of mice sacrificed at day 21 or day 22 post birth. Germfree (N: D14 = 3, D21 = 7, D28 = 8), PedsCom (N: D14 = 4, D21 = 7, D28 = 4), CMCom (N: D14 = 7, D21 = 13, D28 = 5). Independent sample t-test **p<0.01, ***p<0.001. E. Median cecal short-chain fatty acid concentration in germfree, CMCom and PedsCom mice at weaning (day 21) and Oligo-MM12 mice in adulthood (8–14 weeks old). N ≥ 5 per timepoint, per group. GF = germfree, CC = CMCom, PC = PedsCom, OMM12 = Oligo-MM12. F. Median cecal primary bile acid concentration in germfree, CMCom, PedsCom and Oligo-MM12 at weaning [day 21 (N: germfree = 3, CMCom and PedsCom = 6)] and adulthood [8–14 weeks old (N: germfree = 4, CMCom, PedsCom = 5, Oligo-MM12 = 8)] (left panel). The median molar ratios of primary deconjugated to primary conjugated bile acid in each study group. G. Serum IgA concentration in germfree, PedsCom and CMCom 6-12-week-old adult mice (N: germfree = 8, CMCom = 18, PedsCom = 14) (middle and right panels). H. Percent of mucosal and systemic IgA binding to PedsCom, CMCom and Oligo-MM12 fecal microbes from 7-10-week-old mice. (N: RAG2−/− = 4, CMCom, PedsCom = 9, Oligo-MM12 = 8). Mann-Whitney-Wilcoxon test *p<0.05, **p<0.01, ***p<0.001. Data in C and E-H presented with box and whisker plots. See also Figure S4–6.