Supplementary MaterialsImage_1. intestinal and systemic compartments of secondary abiotic and recolonized mice. Secondary abiotic mice were generated by broad-spectrum antibiotic treatment and perorally recolonized by gavage. Subsequently, lymphocytes from small intestinal and colonic lamina propria, MLN and spleen were Vismodegib ic50 isolated, and analyzed by flow cytometry as described in Materials and Methods. The concentrations of CD8+ lymphocytes in the (A) small intestine, (B) colon, (C) MLN and (D) spleen of naive conventional mice (N), secondary abiotic mice (ABx) and mice re-associated with either (Ec), (Lj) or complex intestinal microbiota by FMT on d7 and d28 post-recolonization are depicted. Columns represent means +SD. Significance levels ( 0.05; ?? 0.01; ??? 0.001). Data were pooled from three independent experiments. Image_2.TIFF (929K) GUID:?8B82A53C-294A-4D66-BA3F-1EB4E616AF78 Image_3.TIFF (869K) GUID:?73E46B0E-9F43-4DE4-A994-23147CE9DA9D FIGURE S3 Secondary abiotic mice were generated by broad-spectrum antibiotic treatment and perorally recolonized by gavage. Subsequently, lymphocytes from small intestinal and colonic lamina propria, MLN and spleen were isolated, and analyzed by flow cytometry as described in Materials and Methods. The proportions of CD8+ memory/effector cells (CD8+CD44hi, gated on CD8+ cells) in the (A) small intestine, (B) colon, (C) MLN and (D) spleen of naive conventional mice (N), secondary abiotic Vismodegib ic50 mice (ABx) and mice re-associated with either (Ec), (Lj) or complex intestinal microbiota by FMT on d7 and d28 post-recolonization are depicted. Columns represent means +SD. Significance levels ( 0.05; ?? 0.01; ??? 0.001). Data were pooled from three independent experiments. Image_3.TIFF (869K) GUID:?73E46B0E-9F43-4DE4-A994-23147CE9DA9D Image_4.TIFF (798K) GUID:?27F6D7F9-8167-4A45-86A9-B318D78F7F3D FIGURE S4 Secondary abiotic mice were generated by broad-spectrum antibiotic treatment and perorally recolonized by gavage. Subsequently, lymphocytes from small intestinal and colonic lamina propria, MLN and spleen were isolated, and analyzed by flow cytometry as described in Materials and Methods. The frequencies of activated DCs (CD86+, gated on CD4-CD8- Vismodegib ic50 live cells) in the (A) small intestine, (B) colon, (C) MLN and (D) spleen of naive conventional mice (N), secondary abiotic mice (ABx) and mice re-associated with either (Ec), (Lj) or complex intestinal microbiota by FMT on d7 and d28 post-recolonization are depicted. Columns represent means +SD. Significance levels ( 0.05; ?? 0.01; ??? 0.001). Data were pooled from three independent experiments. Image_4.TIFF (798K) GUID:?27F6D7F9-8167-4A45-86A9-B318D78F7F3D Abstract The essential role of the intestinal microbiota in the well-functioning of host immunity necessitates the investigation of species-specific impacts on this interplay. Aim of this study was to examine the ability of defined Gram-positive and Gram-negative intestinal commensal bacterial species, namely and or with a complex murine microbiota by fecal microbiota transplantation (FMT). Analyses at days (d) 7 and 28 revealed that immune cell populations in the small and large intestines, mesenteric lymph nodes and spleens of mice were decreased after antibiotic treatment but were completely or at least partially restored upon FMT or by recolonization with the respective bacterial species. Remarkably, recolonization resulted in the highest CD4+ and CD8+ cell figures in the small intestine and spleen, whereas neither of the commensal varieties could stably restore those cell populations in the colon until d28. In the mean time less efficient than FMT, both varieties improved the frequencies of regulatory T cells and triggered dendritic cells and completely restored intestinal memory space/effector T cell populations at d28. Furthermore, recolonization with either solitary varieties managed pro- and anti-inflammatory immune functions in parallel. However, FMT could most efficiently recover Vismodegib ic50 the decreased frequencies of cytokine generating CD4+ lymphocytes in mucosal and systemic compartments. recolonization improved the production of cytokines such as TNF, IFN-, IL-17, and IL-22, Rabbit Polyclonal to Aggrecan (Cleaved-Asp369) particularly in the small intestine. Conversely, only recolonization managed colonic IL-10 production. In summary, FMT appears to be most efficient in the repair of antibiotics-induced security damages to the immune system. However, defined intestinal commensals such as and have the potential to restore individual functions of intestinal and systemic immunity. In conclusion, our data provide novel insights into the distinct part of individual commensal.