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目的研究慢性乙型肝炎(CHB)患者外周血单核细胞中T淋巴细胞免疫球蛋白黏蛋白分子3(Tim-3)的表达及对其炎症因子生成的影响。方法利用多色流式技术和荧光实时定量PCR(qRT-PCR),分别检测不同状态的CHB患者外周血单核细胞亚群表面Tim-3分子和Tim-3 mRNA表达水平。给予重组人galectin-9和脂多糖(LPS)共刺激CHB活动期患者外周血单核细胞,用ELISA检测培养上清中TNF-α、IL-1β和IL-6水平。结果慢乙肝免疫耐受期患者(IT)外周血单核细胞表面Tim-3表达水平显著高于健康对照组(HC)和免疫活动期患者(IA)(P<0.01),其中IT患者CD14+CD16-和CD14+CD16+单核细胞亚群中Tim-3的表达均高于IA患者,差异均有统计学意义。qRT-PCR结果进一步证实IA患者外周血单核细胞中Tim-3 mRNA的表达高于HC(P=0.040)。IA患者CD14+单核细胞上Tim-3的表达与血清丙氨酸氨基转移酶(ALT)水平呈正相关(r=0.362,P=0.028)。重组人galectin-9和LPS共刺激IA患者外周血单核细胞比单独使用LPS刺激分泌更高水平的TNF-α,IL-1,IL-6。结论 CHB患者外周血单核细胞Tim-3表达上调,与血清ALT水平呈正相关,Tim-3/galectin-9通路能够促进LPS诱导的单核细胞的炎症因子的分泌。
Objective To study the expression of T-lymphocyte immunoglobulin mucin molecule 3 (Tim-3) in peripheral blood mononuclear cells of patients with chronic hepatitis B (CHB) and its effect on the production of inflammatory cytokines. Methods The expressions of Tim-3 and Tim-3 mRNA on peripheral blood mononuclear cells were detected by qRT-PCR and multi-color flow cytometry. Peripheral blood mononuclear cells were co-stimulated with galectin-9 and lipopolysaccharide (LPS) in patients with active CHB. The levels of TNF-α, IL-1β and IL-6 in culture supernatants were detected by ELISA. Results The expression of Tim-3 on peripheral blood mononuclear cells in patients with chronic hepatitis B (IT) was significantly higher than that in healthy controls (HC) and immune active patients (IA) (P <0.01) The expression of Tim-3 in CD16- and CD14 + CD16 + monocyte subsets was higher than that in patients with IA, the differences were statistically significant. qRT-PCR results further confirmed the expression of Tim-3 mRNA in peripheral blood mononuclear cells of patients with IA was higher than that of HC (P = 0.040). The expression of Tim-3 on CD14 + monocytes in patients with IA was positively correlated with the level of serum ALT (r = 0.362, P = 0.028). Peripheral blood mononuclear cells from recombinant human galectin-9 and LPS co-stimulated IA secreted higher levels of TNF-α, IL-1 and IL-6 than LPS alone. Conclusion The expression of Tim-3 in peripheral blood mononuclear cells of CHB patients is up-regulated, which is positively correlated with serum ALT level. The Tim-3 / galectin-9 pathway can promote the LPS-induced secretion of inflammatory cytokines in monocytes.