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. Author manuscript; available in PMC: 2021 Aug 18.
Published in final edited form as: Immunity. 2020 Aug 18;53(2):398–416.e8. doi: 10.1016/j.immuni.2020.07.010

Figure 5. Paneth cell lysozyme deficiency or overproduction alters gut microbiota composition.

Figure 5.

(A) Bacterial loads were determined in ileal and colonic mucosa and cecal lumen in WT and Lyz1−/− mice (N=6–10 in each group). (B-H) 16S amplicon profiling of fecal or ileal luminal microbiota from WT and Lyz1−/− mice. (B) Chao1 index (alpha diversity) (fecal microbiota, n=10– 12). (C) Unweighted UniFrac analysis of fecal microbiota from WT and Lyz1−/− mice (n=14–16). (D-E) PCoA analysis of fecal microbiota with consideration for genotype and cage effect (total of 8 WTs in 3 cages; 10 Lyz1−/−s in 4 cages). (F) Mixed linear model analysis using genotype as fixed and cage as random effect. One-way ANOVA showed that cage effects contributed to differences among mice of the same genotype. (G) PCoA analysis of ileal luminal microbiota from separately housed adult Lyz1−/− and WT littermates. (H) LDA of ileal luminal bacteria with species contracting (Candidatus Arthromitus) or expanding (R. gnavus, B. gnavus, and D. formicigenerans) in Lyz1−/−mice. (I) Schematic diagram of the Villin-Lyz1TG transgenic (TG) construct. (J) Lysozyme immunohistochemistry for WT, Lyz1−/−, and TG mouse colons (representative of N>3 for each genotype). (K) Lysozyme enzymatic activity in the colonic lumen of WT, Lyz1−/−, and TG mice (N=3 for each genotype). (L) Unweighted UniFrac analysis (16S amplicon profiling) of WT and TG mouse fecal microbiota. (M) Relative phyla abundance in the feces of separately housed WT and Lyz1−/− mice (N=4–8). (N) Relative phyla abundance of WT and TG mice (N=5–6). (O) Relative abundance of R. gnavus and D. formicigenerans in WT and Lyz1−/− fecal microbiota (N=4–10 per genotype). (P) Relative abundance of R. gnavus and D. formicigenerans in WT and TG fecal microbiota (N=5–6). (Q-V) Growth sensitivity of selected bacteria to lysozyme based on OD 660nm readings. Red arrowhead indicated the addition of 200 μg/ml lysozyme (data from 2–4 independent experiments). All bar graphs display mean ± SEM from at least two independent experiments. See also Figure S5.