小鼠膀胱组织中驻留αβΤ和γδΤ细胞表型和细胞因子表达的探讨

Investigation of phenotype and cytokine expression of resident αβΤ and γδΤcells in murine bladder tissues

目的:探讨小鼠膀胱组织中T细胞的表型和细胞因子表达。方法:获取小鼠膀胱单个核细胞,流式细胞术检测小鼠膀胱组织中T细胞的比例、趋化因子受体和细胞因子的表达。HE染色观察膀胱结构,免疫组化和免疫荧光观察正常小鼠与MB49原位肿瘤小鼠膀胱组织中CD3^(+)T和γδΤ细胞的分布。结果:正常小鼠膀胱中含有组织驻留记忆T细胞,表达记忆分子CD44和组织驻留标志分子CD69和CD103,其中γδΤ细胞的比例显著高于外周血并高表达趋化因子受体CXCR3和细胞因子IL-17、TNF-α。与正常膀胱组织相比,MB49原位肿瘤小鼠膀胱组织中CD4^(+)T细胞显著降低,CD8^(+)T和γδΤ细胞比例显著升高,并高表达IFN-γ。此外,γδΤ细胞还显著高表达IL-17和TNF-α。结论:小鼠膀胱组织中具有组织驻留记忆αβΤ和γδΤ细胞,且表达细胞因子IFN-γ,IL-17和TNF-α,为进一步深入研究膀胱癌奠定基础。

Objective:To explore the phenotype and cytokine expression of resident T cells in murine bladder tissues.Methods:The percentages of T cells and the expression of chemokine receptors and cytokines in bladder tissues were analyzed by flow cytometry. HE staining was used to confirm the structure of bladder and the distribution of CD3^(+)T and γδΤ cells in mouse models of normal and orthotopic bladder tumor were determined by immunohistochemical and immunofluorescence.Results:The majority of T cells in bladder tissues were tissue-resident memory T cells,highly expressing memory marker CD44 and residence marker CD69 and CD103. The proportion of γδΤ cells in bladder tissues was significantly higher than that in blood,which dramatically expressed chemokine receptor CXCR3 and cytokines IL-17 and TNF-α. Compared to normal bladder tissues,the proportion of CD4^(+)T cells in bladder with MB49-induced tumor has significantly reduced. In contrast,the proportion of CD8^(+)T cells and γδΤ cells has significantly increased in bladder with MB49-induced tumor. Furthermore,CD4^(+)T cells,CD8^(+)T cells and γδΤcells in bladder with MB49-induced tumor expressed higher levels of IFN-γ. In addition,there was also increased in the expression of IL-17 and TNF-α by γδΤ cells in bladder with MB49-induced bladder tumors than that in control bladder tissues.Conclusion:Most of the αβΤ and γδΤ cells in bladder tissues were tissue-resident memory T cells,which remarkably expressed IFN-γ,IL-17 and TNF-α and might play a critical role in anti-tumor processes. Our study will lay the foundation for further research on the treatment of bladder cancer.