目的观察高迁移率族蛋白B1(HMGB1)诱导的树突状细胞(DCs)对大鼠脾T淋巴细胞增殖反应及功能性极化的影响,并探讨其作用机制。方法分离正常Wistar大鼠脾脏DC后置于24孔培养板(1×10^6/孔),采用1mg/L的HMGB1刺激48h,DC与脾T淋巴细胞的细胞比例分别为1:50、1:100、1:150、1:200混合,于共同作用后3、5d观察脾T淋巴细胞增殖及功能性极化的情况。结果DC与T淋巴细胞以1:50、1:100、1:150、1:200的比例相互作用,脾T淋巴细胞对丝裂原刺激的增殖反应均明显增强、上清液中干扰素(IFN)-γ含量显著升高和白细胞介素(IL)-4含量明显降低(P〈0.01),其中细胞比例1:150组最为明显。脾T淋巴细胞对丝裂原刺激的增殖反应增强、IFN-γ含量增加和1L4含量降低均以第3天最为显著[分别为(0.646±0.028)、(268.08±14.07)、(6.16±0.59)ng/L,P值均〈0.01]。结论HMGB1诱导的DC使脾T淋巴细胞对丝裂原刺激的增殖反应明显增强,并促进T淋巴细胞向Th1功能性极化。
Objective:To investigate the effects of splenic dendritic cells (DCs) treated with high mobility group box-1 protein (HMGB1) on T lymphocytes proliferation and polarization and its potential mechanism in rats. Methods DCs isolated from the spleen in male Wistar rats were seeded on 24-well ( 1 ×10^6 cells/well) cell culture plates, and the cells were stimulated with HMGB1 in a concentration of 1 mg/L for 48 h. After being stimulated,DCs were co-cultured with T lymphocytes at different ratios ( 1: 50, 1: 100,1: 150,and 1:200). The cells were collected on day 3 and 5 after culture to determine T lymphocytes proliferation and polarization. Results Splenic T lymphoeytes proliferative response to T cell mitogen, concanavalin A (Con A ) , were significantly increased on day 3 and 5 after cultured in the presence of DC to T lymphocyte ratios in 1 : 50,1 : 100,1 : 150 and 1 : 200 ( all P 〈 0.01 ), with marked production of intefferon-У and reduction in interleukin-4 in supernatants ,especially when DC to T lymphocytes ratio was 1:150 and cultured for 3 days [ interferon-У and interleukin-4 levels: (268.08 - 14.07) and (6.16 - 0.59) ng/L,respectively, all P 〈 0.01 ]. Condusion Splenic dendritic cells treated with HMGB1 can result in a marked increase in T lymphocytes proliferation, and induce T lymphocytes differentiation to Thl.