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目的:体外研究不同浓度拉米夫定(lamivu- dine,LAM)对慢性乙型肝炎(chronic hepatitis B,CHB)患者树突状细胞(dendritic cell,DC)功能影响.方法:分离慢乙肝患者外周血单核细胞,在含GM-CSF+IL-4及不同浓度LAM(0,0.125, 0.25,0.5,1,2 mmol/L)培养条件下制备DC.观察DC形态学变化并用MTT法测定DC刺激同种异体淋巴细胞增殖能力,流式细胞仪(FCM) 测定其细胞表型分子CD1a,CD83,CD80及 HLA-DR的表达,ELISA法检测培养上清中 IL-12和IL-6含量.结果:在LAM 0.5 mmol/L组,DC表型分子 CD83,CD1a,HLA-DR的表达最高,CD80与 LAM未处理组相比无明显差异.与LAM未处理组相比,LAM 0.5 mmol/L处理组DC膜表面分子CD1a,CD83,HLA-DR表达增高(CD1a: 54.0±9.2 vs 33.6±10.1,P<0.05;CD83:20.3 ±6.1 vs 11.8±6.2,P<0.05;HLA-DR:74.5± 7.1 vs 52.9±7.7,P<0.05);其上清液中IL-12 分泌水平增高(810.0±91.5 ng/L vs 268.0± 34.3 ng/L,P<0.05),IL-6则显著降低(28.1± 2.6 ng/L vs 55.3±7.4 ng/L,P<0.05);刺激同种异体淋巴细胞增殖能力(SI)增强(1.9±0.6 vs 1.2±0.5,P<0.05).结论:LAM体外可增强慢乙肝患者树突状细胞活性.
Objective: To investigate the effect of lamivuidine (LAM) in different concentrations on the dendritic cell (DC) function in patients with chronic hepatitis B (CHB) in vitro. Methods: Peripheral blood mononuclear cells from patients with chronic hepatitis B were isolated and cultured with GM-CSF + IL-4 and LAM (0,0.125, 0.25,0.5,1,2 mmol / L) DC was prepared. The morphological changes of DCs were observed and the proliferation of allogeneic lymphocytes stimulated by DC was measured by MTT assay. The expression of CD1a, CD83, CD80 and HLA-DR were detected by flow cytometry (FCM) In IL-12 and IL-6 content. Results: In LAM 0.5 mmol / L group, the expression of CD83, CD1a and HLA-DR was the highest, but there was no significant difference between CD80 and LAM untreated group. Compared with LAM untreated group, the expression of CD1a, CD83 and HLA-DR on DC membrane in LAM 0.5 mmol / L group increased (CD1a: 54.0 ± 9.2 vs 33.6 ± 10.1, P <0.05; CD83: 20.3 ± 6.1 vs 11.8 ± 6.2, P <0.05; HLA-DR: 74.5 ± 7.1 vs 52.9 ± 7.7, P <0.05). IL-12 secretion in the supernatant increased (810.0 ± 91.5 ng / L vs 268.0 ± 34.3 ng / L, P <0.05) (28.1 ± 2.6 ng / L vs 55.3 ± 7.4 ng / L, P <0.05). The proliferation of allogeneic lymphocytes stimulated (1.9 ± 0.6 vs 1.2 ± 0.5, P <0.05). Conclusion: LAM can enhance dendritic cell activity in patients with chronic hepatitis B in vitro.