肝星状细胞通过诱导产生CD8^+CD28^-T淋巴细胞抑制T细胞增殖  

Hepatic Stellate Cells Inhibit Proliferation of T Cells through Induction of CD8^+CD28^-T Lymphocytes

作　　者：刘秋莉[1] 刘畅[1] 杨帆[1] 郑海清[2] LIU Qiu-li LIU Chang YANG Fan ZHENG Hai-qing(Cell-gene Therapy Translational Medicine Research Center Department of Rehabilitation Medicine, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou510630, China)

机构地区：[1]中山大学附属第三医院生物治疗中心,广东广州510630 [2]中山大学附属第三医院康复医学科,广东广州510630

出　　处：《中山大学学报（医学科学版）》2016年第6期817-821,共5页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金(81572228;81601381);广东省自然科学基金(2015A030313015;2016A030310131);中山大学高校青年教师培育项目(15ykpy26);广东省科技计划项目(2014A020212715)

摘　　要：【目的】研究肝星状细胞(HSC)是否通过诱导产生CD8+CD28-T淋巴细胞抑制T细胞增殖。【方法】流式细胞仪分选出CD3+T细胞,用羧基荧光素双乙酸盐琥珀酰亚胺酯(CFSE)标记后与人肝星状细胞系LX2细胞共培养,PHA刺激72 h后,流式细胞仪检测T细胞增殖的百分比;为了研究肝星状细胞调控T细胞增殖的机制,将CD3+T细胞与LX2细胞共培养3 d,流式细胞仪检测CD8+CD28-T淋巴细胞的百分比。进一步将CD3+T细胞与LX2细胞共培养3 d,通过流式细胞仪分选出CD8+CD28-T淋巴细胞与CFSE标记的CD3+T细胞共培养,PHA刺激72 h后,流式细胞仪检测T细胞增殖的百分比。【结果】LX2细胞明显抑制了T细胞增殖,且抑制作用呈剂量依赖性;LX2剂量依赖性上调CD8+CD28-T淋巴细胞百分比。和LX2细胞共培养后的CD8+CD28-T淋巴细胞能够显著抑制T细胞增殖。【结论】HSC细胞通过诱导CD8+CD28-T调节细胞比例抑制T淋巴细胞增殖。[Objective] To investigate whether hepatic stellate cells （HSC） inhibit the proliferation of T cells through the induction of CD8^＋ CD28^- T lymphocytes. [ Methods ] CD3^＋ T cells were sorted by flow cytometry and labeled by 5,6-carboxyfluoresceindiacetate, succinimidyl ester （CFSE） , then they were co-cultured with human hepatic stellate cells cell line LX2 in the present of PHA for 72 h. The proliferation of T cells was evaluated by flowcytometry.In order to study the mechanism of the inhibition of T cells by hepatic stellate ceils, purified CD3^＋ T cells were co-cultured with LX2 for 3 days, and the percentage of CD8^＋ CD28^- T cells was detected by flowcytometry. Furthermore, after CD3^＋ T cells were co-cultured with LX2 cells for 3 days, CD8^＋ CD28^- T cells were sorted by flow cytometry. Then purified CD8^＋ CD28^- T cells were co-cultured with CFSE labeled CD3^＋ T cells, and the proliferation of T cells was evaluated after stimulated with PHA for 72 h. [ Results ] T cell proliferation was inhibited obviously after co-cultured with LX2 cells. The inhibitory effect was dose dependent. LX2 cells dose-dependently increased the percentage of CD8^＋ CD28^- T cells. CD8^＋ CD28^-F cells, which cultured with LX2 cells, obviously inhibited the proliferation of T cells. [Conclusion] HSC inhibited the T cell proliferation through the induction of CD8^＋ CD28^- T regulatory cells.

关 键 词：肝星状细胞 T细胞 CD8+CD28-T淋巴细胞 

分 类 号：R392[医药卫生—免疫学]