小鼠肠道3型固有淋巴样细胞检测与分析方法研究

目的建立小鼠肠道(包括小肠和结肠)黏膜固有层3型固有淋巴样细胞(ILC3)检测与分析的方法。方法小鼠麻醉后分离小肠和结肠组织,利用不同的消化液分别获得小肠和结肠固有层淋巴细胞悬液,通过CD45⁺CD3^-CD90.2⁺RORγt⁺流式分析策略检测ILC3比例,通过IL-23刺激后用流式细胞术分析显示ILC3的IL-22表达水平。结果小肠和结肠固有层淋巴细胞成功分离,流式细胞术检测显示ILC3细胞分群显著并明显表达IL-22。在小肠和结肠中,ILC3在淋巴细胞中的占比分别为(8.30±2.42)%和(1.43±0.34)%,IL-22⁺ILC3在RORγt⁺ILC3中的占比分别为(38.98±8.76)%和(85.86±1.72)%。结论本研究成功建立了小鼠小肠和结肠黏膜固有层ILC3的检测和分析方法,为相关研究奠定了技术基础。

Detection and analysis method of group 3 innate lymphoid cells in mice intestine

Objective To establish a method for the detection and analysis of group 3 innate lymphoid cells(ILC3)in the lamina propria of intestinal mucosa(including the small intestine and colon)in mice.Methods After the mice were anesthetized,the small intestine and colon tissues were separated,and the lymphocyte suspensions in the lamina propria of small intestine and colon were obtained from different digestive solutions.The proportion of ILC3 was detected through CD45⁺CD3^-CD90.2⁺RORγt⁺flow cytometry strategy.After stimulated with IL-23,the IL-22 expression level in ILC3 was detected by flow cytometry.Results The lymphocytes in the lamina propria of small intestine and colon were successfully isolated,and flow cytometry showed that the ILC3 was significantly clustered and IL-22 was significantly expressed.In the small intestine and colon,the proportion of ILC3 in lymphocytes was(8.30±2.42)%and(1.43±0.34)%,and the proportion of IL-22⁺ILC3 in RORγt⁺ILC3 was(38.98±8.76)%and(85.86±1.72)%,respectively.Conclusion This study successfully established the detection and analysis method of ILC3 in the lamina propria of small intestine and colon mucosa in mice,which provides a technical foundation for the related research.