Supplementary MaterialsSupplemental Shape 1: GF-AF mice in this study have a similar phenotype to those in a previous report. 6), GF (= 6), and GF-AF mice (= 3). (ACE) Data are pooled from at least three impartial experiments. 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 impartial experiments. (B) Representative flow cytometry plots of IgA vs. B220 Sofosbuvir impurity A on CD3? lymphocytes of SI-LP and LI-LP of SPF and SPF-AF 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 impartial 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 impartial experiments. All data are mean SD. Welch's test was performed for statistical analysis (E,F). Sofosbuvir impurity A *< 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 impartial 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 impartial 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 Physique 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 amounts in SPF-AF and SPF mice. Mature ILFs had been counted by calculating how big is the B220+ region, and if 50,000 m2, the ILFs had been characterized as mature. The amounts of older and total ILF were counted in the next elements of the mouse intestine; (A,E) Proximal SI. (B,F) Distal SI. (C,G) Top fifty percent of LI. (D,H) Decrease fifty percent of LI. Sofosbuvir impurity A The intestinal regions were thought as referred to in the techniques and Components Mouse monoclonal to CD19 section. Data are pooled from two indie tests (= 4). Mean SD. are proven. Welch’s < 0.01. Picture_5.jpeg (404K) GUID:?0A85D642-6411-4442-8B2A-F50A1E6EAF21 Abstract The principal induction sites for intestinal IgA will be the gut-associated lymphoid tissue (GALT), such as for example Peyer's patches (PPs) and isolated lymphoid follicles (ILFs). The commensal microbiota may donate to IgA creation in the gut; nevertheless, the role of dietary antigens in IgA production is understood poorly. To comprehend the result of nutritional antigens on IgA creation, 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 cells and germinal center B cells. The role of dietary antigens in the PP responses was further confirmed by adding bovine serum albumin (BSA) into the elemental diet. Although dietary antigens are important Sofosbuvir impurity A for PP responses, they have fewer effects than the microbiota around the development and maturation of ILFs. Furthermore, we exhibited that dietary antigens are essential for a normal antigen-specific IgA response to serovar Typhimurium contamination. These results provide new insights into the role of dietary antigens in the regulation Sofosbuvir impurity A of mucosal immune responses. serovar Typhimurium. Results Dietary Antigens Contribute to IgA Production in SI but Not in LI To assess the functions of dietary antigens in the intestinal immune system, we prepared an antigen-free (AF) diet and established.