CFSE标记结合流式细胞术检测单核细胞Siglec-1刺激CD4^+/CD8^+T细胞增殖

Combined application of CFSE labeling and flow cytometry on the detection of the proliferation of CD4^+/CD8^+ T cells simulated with monocytes Siglec-1

目的建立CFSE标记结合流式细胞术检测T淋巴细胞增殖的方法,并探讨其在研究单核细胞唾液酸黏附素(Si-glec-1)对CD4+/CD8+T细胞增殖中的应用。方法免疫磁珠分离冠状动脉综合征(ACS)患者和健康对照者外周血CD14+单核细胞,与经CFSE标记的第三方健康献血者CD4+/CD8+T细胞共培养5 d,流式细胞术检测T淋巴细胞增殖情况。结果 ACS患者CD14+单核细胞具有比健康人单核细胞更强的刺激CD4+/CD8+T淋巴细胞增殖能力;当用干扰素-α(INF-α)刺激增强正常单核细胞Siglec-1表达后,其刺激T细胞增殖能力增强;用单抗阻断单核细胞Siglec-1后,其刺激T细胞增殖能力减弱。结论 ACS患者增强的CD4+/CD8+T淋巴细胞增殖能力部分是由于激活的单核细胞高表达Siglec-1引起。

Objective To establish a combined detection method of CFSE labeling and flow cytometry （FCM） for the measure ment of the proliferation of T lymphocytes and to explore its role for the investigation of the effect of Siglec 1 in monocytes on the proliferation of CD4^＋/CD8^＋ T ceils. Methods CD14^＋ monocytes in peripheral blood of patients with acute coronary syndrome （ACS） and healthy controls were isolated by magnetic-activated cell sorting （MACS）. CD4^- and CD8^＋ T cells from beahhy blood donors of third-party were also separated by MACS and labeled with CFSE. 5 days after the mixed culture of monocytes and lyre pbocytes,the proliferation of T lymphocyte was determined by FCM. Results CD14^＋ monocytes from patients with ACS had a more robust role in stimulating the proliferation of CD4^＋/CD8^＋ T lymphocyte than those from healthy controls. However, when Si glec 1 on monocytes was blocked by anti Siglec-1 antibody,the stimulating capacity was decreased to normal level. Furthermore,af ter being stimulated with INF-α,the expression of Siglec-1 on monocytes from healthy controls was elevated and the stimulating ca pacity of monocytes was enhanced. Conclusion The increased proliferation of CD4＋/CDS- T lymphocyte in patients with ACS was partly due to the elevated expression of Siglec 1 on activated monocytes.