One-way ANOVA with Tukey’s test was performed for statistical analysis. IgA concentration in feces from SPF, GF, and GF-AF mice. (G,H) The absolute numbers of B220-IgA+ IgA-producing plasma cells in SI-LP (G) and LI-LP (H) of SPF (= 6), GF (= 6), and GF-AF mice (= 3). (ACE) Data are pooled from at least three independent experiments. Mouse monoclonal to LPA Data are mean SD. One-way ANOVA with Tukey’s test was performed for statistical analysis. * 0.05, ** 0.01, *** 0.001, **** 0.0001. Image_1.jpeg (252K) GUID:?20A4C5EA-E177-43D7-8A87-72242F15D1E6 Supplementary Figure 2: IgA production and changes in the fecal microbiota composition of SPF-AF mice. (A) IgA concentration in feces from SPF (= 6) and SPF-AF mice (= 6). Data are pooled from two independent experiments. (B) Representative flow cytometry plots of IgA vs. B220 on CD3? lymphocytes of SI-LP and LI-LP of SPF and SPF-AF Abiraterone Acetate (CB7630) mice (left), with the absolute numbers of B220?IgA+ IgA-producing plasma cells (right). (C,D) Microbiota compositions of SPF mice (= 5) and SPF-AF (= 5) mice are shown at phylum level (C) and genus level (D). Data are presented as mean SD and Welch’s 0.01, *** 0.001, **** 0.0001. Image_2.jpeg (514K) GUID:?88A6351B-191D-4EA3-8B4B-7FC66DC60BF8 Supplementary Figure 3: Dietary antigens affect GC B cells and Tfh cells in PP and MLN. (A,B) The number of leukocytes (A) and GC B cells (B) in PP of GF-AF mice and GF-AF mice fed AF diet supplemented with 1% BSA. (C,D) The number of GC B (B220+CD19+ Fas+GL7+) cells (C) and Tfh (CD19?CD3+CD4+CXCR5+PD-1+) cells (D) in PP of SPF (= 4 or 6) and SPF-AF (= 4 or 6) mice. Data are pooled from at least two independent experiments. (E,F) The number of GC B cells (E) and Tfh cells (F) in MLN of SPF (= 7 or 8), GF (= 8 or 9), and GF-AF (= 8 or 9) mice. Data are pooled from at least two independent experiments. All data are mean SD. Welch’s test was performed for statistical analysis (E,F). * 0.05, ** 0.01, **** 0.0001. Image_3.jpeg (269K) GUID:?0DF28590-B049-4736-BF7D-ABA6FBEAB8E6 Supplementary Figure 4: Nrp-1?RORt? pTreg cells in PP are reduced in GF-AF mice. (A) The number of Neuropilin-1low RORt? Foxp3+ CD4 T cells in PP of SPF (= 4), GF (= 4), and GF-AF (= 5) mice. Data are representative of two independent experiments. (B) The number of IL-17A producing CD4 T cells in PP of SPF (= 4), GF (= 3), and GF-AF (= 3) mice. Data are pooled from two independent experiments. All data are mean SD. One-way ANOVA with Tukey’s test was performed for statistical analysis. * 0.05, ** 0.01, **** 0.0001. Image_4.jpeg (246K) GUID:?BE40C5B0-528E-463B-B131-CFBBF8B26A0D Supplementary Figure 5: The development and maturation of ILF are altered by dietary antigen through the microbiota in some parts of the intestine. (ACD) Total ILF numbers; (ECH) Mature ILF numbers in SPF and SPF-AF mice. Mature ILFs were counted by measuring the size of the B220+ area, and if 50,000 m2, the ILFs were characterized as mature. The numbers of total and mature ILF were counted in the following parts of the mouse intestine; (A,E) Proximal SI. (B,F) Distal SI. (C,G) Upper half of LI. (D,H) Lower half of LI. The intestinal regions were defined as described in the Materials and Methods section. Data are pooled from two independent experiments (= 4). Mean SD. are shown. Welch’s 0.01. Image_5.jpeg (404K) GUID:?0A85D642-6411-4442-8B2A-F50A1E6EAF21 Abstract The primary induction sites for intestinal IgA are the gut-associated lymphoid tissues (GALT), such as Peyer’s patches (PPs) and isolated lymphoid follicles (ILFs). The commensal microbiota is known to contribute to IgA production in the gut; however, the role of dietary antigens in IgA production is poorly understood. To understand the effect of dietary antigens on IgA production, post-weaning mice were maintained on an elemental diet without any large immunogenic molecules. We found that dietary antigens contribute to IgA production in PPs through induction of follicular helper T Abiraterone Acetate (CB7630) cells and germinal Abiraterone Acetate (CB7630) center B cells. The role of dietary antigens in the PP responses was further confirmed by adding bovine serum albumin (BSA).