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目的:用免疫荧光染色和流式细胞术(FCM)鉴定正常人外周血单个核细胞(PBMC)中滤泡辅助性T(Tfh)细胞,以便对其功能进行进一步研究。方法:密度梯度离心法分离人PBMC,采用不同荧光素标记的抗CXCR5、CD4及CD19抗体进行染色FCM分析,鉴定出表型为CD4+CXCR5+的T细胞亚群。结果:外周血可以检测到CD4+CXCR5+T细胞亚群,占CD4+T淋巴细胞百分比为(11.09±0.38)%,MFI值为25.78±0.72,低于B细胞膜表面CXCR表达水平(MFI值为99.6±8.1)。结论:健康人外周血存在一定比例的Tfh细胞,应用FCM可以成功鉴定Tfh细胞,为进一步研究其功能提供了良好的技术平台。
OBJECTIVE: To identify follicular helper T (Tfh) cells in peripheral blood mononuclear cells (PBMCs) of normal people by immunofluorescence staining and flow cytometry (FCM) to further study their function. Methods: Human peripheral blood mononuclear cells (PBMCs) were isolated by density gradient centrifugation. FCM analysis was performed using different fluorescein-labeled anti-CXCR5, CD4 and CD19 antibodies to identify T cell subsets with phenotypes of CD4 + CXCR5 +. Results: The CD4 + CXCR5 + T cell subsets were detected in peripheral blood, the percentage of CD4 + T lymphocytes was (11.09 ± 0.38)%, the MFI was 25.78 ± 0.72, which was lower than the expression of CXCR on B cell membrane 99.6 ± 8.1). Conclusion: There is a certain proportion of Tfh cells in peripheral blood of healthy people. FCM can successfully identify Tfh cells, which provides a good technical platform for further study on its function.