Th17-mediated mucosal inflammation is related to increased Prevotella bacterial abundance. The actual involvement of Prevotella in the development and accumulation of intestinal Th17 cells at a steady state, however, remains undefined. Herein, we investigated the role of Prevotella in inducing intestinal Th17 cells in mice. Mice were treated with a combination of broad-spectrum antibiotics (including ampicillin, neomycin sulfate, vancomycin hydrochloride, and metronidazole) in their drinking water for 4 weeks and then gavaged with Prevotella for 4 weeks. After inoculation, 16S rDNA sequencing was used to verify the colonization of Prevotella in the colon of mice. The IL-17A as well as IL-17A-expressing T cells was localized and quantified by an immunofluorescence assay (IFA) of colon sections. Th17 cells in the mesenteric lymph nodes of mice were counted by flow cytometry. Systemic immune response to Prevotella colonization was evaluated based on the serum levels of IL-6, TNF-α, IL-1β, IL-17A, IL-10, IL-4, IFN-γ, and IL-2. Th17-polarizing cytokines (IL-6, TNF-α, IL-1β, and IL-2) induced by Prevotella were evaluated by stimulation of bone marrow-derived dendritic cells (BMDCs). Results revealed that after inoculation, Prevotella successfully colonized the intestine of mice and induced the production and accumulation of colonic Th17 cells in the colon. Moreover, Prevotella elevated some of the Th17-related cytokines in the serum of mice. And Th17-polarizing cytokines (IL-6 and IL-1β) produced by BMDCs were mediated mainly through the interaction between Prevotella and Toll-like receptor 2 (TLR2). In conclusion, our data suggest that Prevotella induces the production of Th17 cells in the colon of mice, thus highlighting the potential role of Prevotella in training the intestinal immune system.

中文翻译：

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普氏菌诱导小鼠结肠产生 Th17 细胞

Th17 介导的粘膜炎症与普氏菌丰度增加有关。然而，普雷沃氏菌在稳定状态下参与肠道 Th17 细胞的发育和积累的实际情况仍未确定。在此，我们研究了普氏菌在诱导小鼠肠道 Th17 细胞中的作用。小鼠在饮用水中联合使用广谱抗生素（包括氨苄青霉素、硫酸新霉素、盐酸万古霉素和甲硝唑）治疗 4 周，然后灌胃普氏菌4 周。接种后，16S rDNA测序验证普氏菌定植在小鼠的结肠中。IL-17A 以及表达 IL-17A 的 T 细胞通过结肠切片的免疫荧光测定 (IFA) 进行定位和定量。流式细胞术对小鼠肠系膜淋巴结中的Th17细胞进行计数。基于 IL-6、TNF- α、IL- 1β、IL-17A、IL-10、IL-4、IFN- γ和 IL-2的血清水平评估对普氏菌定植的全身免疫反应。普雷沃氏菌诱导的Th17 极化细胞因子（IL-6、TNF- α、IL-1 β和 IL-2）通过刺激骨髓衍生树突细胞 (BMDC) 进行评估。结果显示，接种后，普氏菌成功定植小鼠肠道并诱导结肠中结肠 Th17 细胞的产生和积累。此外，普雷沃氏菌提高了小鼠血清中一些与 Th17 相关的细胞因子。BMDCs 产生的 Th17 极化细胞因子（IL-6 和 IL- 1β）主要通过普氏菌和 Toll 样受体 2（TLR2）之间的相互作用介导。总之，我们的数据表明普氏菌诱导小鼠结肠中 Th17 细胞的产生，从而突出了普氏菌在训练肠道免疫系统中的潜在作用。