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

【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.