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目的:用免疫荧光染色和流式细胞术(FCM)分析鉴定人滤泡辅助性T(Tfh)细胞,对外周血和扁桃体来源的Tfh上CD226分子的表达格局进行研究,采用免疫组化染色的方法,检测异位淋巴组织来源的生发中心中CD226分子的表达情况。方法:采用密度梯度离心法分离人外周血PBMC;扁桃腺经研磨、200目滤网过滤获取扁桃腺细胞,对两种细胞采用不同荧光素标记的抗CXCR5、CD4及CD226抗体进行染色和FCM分析。常规制备类风湿性关节炎(RA)关节滑膜活检组织石蜡切片,采用免疫组化的方法分析CD226分子的表达格局。结果:外周血及扁桃腺Tfh细胞上可以检测到CD226分子,外周血Tfh上CD226阳性率为(91.88±0.52%),MFI值为201.49±6.2;扁桃腺Tfh上CD226表达阳性率为(90.81±0.69%),MFI值为230.81±8.4,表达密度稍高于其他CD4+T细胞。RA患者关节滑膜组织可以观察到异位生发中心(GC),并检测到CD226分子的表达。结论:健康人外周血、扁桃腺及RA患者关节滑膜组织中存在不同比例的Tfh细胞,其膜上CD226分子的表达具有独特的格局,该结果为进一步研究其功能提供了思路。
OBJECTIVE: To identify human follicular helper T (Tfh) cells by immunofluorescence staining and flow cytometry (FCM) analysis and to study the expression pattern of CD226 on Tfh from peripheral blood and tonsil. Immunohistochemical staining , Detect the expression of CD226 in the germinal center of ectopic lymphoid tissue. Methods: Peripheral blood PBMCs were isolated by density gradient centrifugation. Amygdala was collected by filtration through a 200-mesh filter, and tonsil cells were obtained. Staining and FCM analysis were performed on the two kinds of cells using different fluorescein-labeled anti-CXCR5, CD4 and CD226 antibodies. Routine preparation of rheumatoid arthritis (RA) synovial biopsy tissue paraffin sections, immunohistochemical analysis of CD226 expression pattern. Results: CD226 was detected on Tfh cells in peripheral blood and tonsil. The positive rate of CD226 on Tfh was (91.88 ± 0.52)% and MFI was 201.49 ± 6.2 in peripheral blood and tonsil. The positive rate of CD226 on Tfh in tonsil was (90.81 ± 0.69%), the MFI value was 230.81 ± 8.4, the expression density was slightly higher than other CD4 + T cells. Synovial tissue from patients with RA can be observed ectopic germinal center (GC), and CD226 molecules were detected. CONCLUSION: Tfh cells with different proportions exist in the synovial tissues of healthy human peripheral blood, tonsil and RA patients. The expression of CD226 on the membrane of the Tfh cells has a unique pattern. This result provides a new idea for further study of its function.