肝细胞癌患者 Treg 细胞对 CD8＋T 淋巴细胞穿孔素表达的影响

摘要

Objective To investigate the influence of immunosuppressive regulatory T cells (Treg)in patients with hepatocellular carcinoma (HCC)on perforin expression of CD8 + T lymphocytes.Methods Perforin expression of CD8 + T lymphocytes in 20 healthy subjects and 20 HCC patients was detected by flow cytometry.CD8 + T lymphocytes from 20 healthy subjects and Treg from 20 HCC patients were collected by magnetic activated cell sorting (MACS),then were divided into two groups:single culture of CD8 + T lymphocytes and co-culture of CD8 + T lymphocytes and Treg.Both groups were stimulated by anti-CD3/CD28 for 48 hours.Perforin expressions of CD8 + T lymphocytes in both groups were quantitatively detected by flow cytometry.Results Perforin expression of CD8 + T lymphocytes in HCC patients (10.74%±3.96%)was similar with that in healthy subjects (12.6% ±2.48%)(P >0.05 ).However,perforin expression was significantly decreased after CD8 + T lymphocytes were co-cultured with Treg [(34.2%±3.65%)and (20.43%±4.52%), respectively (P <0.01)].Conclusion Treg could significantly reduce perforin expression of CD8 + T lymphocytes.%目的：研究肝细胞癌患者免疫抑制性 Treg 细胞对 CD8＋ T 淋巴细胞穿孔素表达的影响。方法采集20例肝细胞癌患者和20名健康人的外周血，用流式分析法检测抗-CD3／CD28刺激48 h 后 CD8＋ T 淋巴细胞的穿孔素表达情况。免疫磁珠分离法分离健康人 CD8＋ T 淋巴细胞和肝细胞癌患者 Treg 细胞，一组 CD8＋ T 淋巴细胞单独培养，另一组CD8＋T 淋巴细胞与 Treg 细胞共同培养，用流式细胞法检测抗-CD3／CD28刺激48 h 后两组 CD8＋ T 淋巴细胞穿孔素表达情况。结果肝细胞癌患者外周血中 CD8＋ T 淋巴细胞穿孔素表达量与健康人相近，分别为（10．74±3．96）％和（12．6±2．48）％，差异无统计学意义（P ＞0．05）。CD8＋ T 淋巴细胞单独培养和与肝癌患者 Treg 细胞共培养后，健康人 CD8＋ T 淋巴细胞穿孔素表达量分别为（34．2±3．65）％和（20．43±4．52）％，差异有统计学意义（t＝11．42，P ＜0．01）。结论肝癌患者免疫抑制性 Treg 细胞可使 CD8＋ T 淋巴细胞穿孔素表达量降低。