拉米夫定体外对慢性乙型肝炎患者树突状细胞功能的影响

In vitro effects of lamivudine on function of dendritic cells derived from patients with chronic hepatitis B infection

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．

AIM： To investigate the effects of lamivudine on the function of dendritic cells derived from patients with chronic hepatitis B （CHB） in vitro. METHODS： Dentritic cells （DCs） derived from the monocytes of CHB patients were cultured with interleukin4 （IL-4） plus granulocyte-macrophage colony-stimulating factor （GM-CSF）. Three days after being cultured, the cells were divided group A and B. Group A was treated with different concentrations of lamivudine （0, 0.125, 0.25, 0.5, 1, 2 mmol/L）, while group B served as the control. Cell morphology was observed under light microscope and cell surface molecules including HLA-DR, CD80, CD83, and CD1a were assayed by flow cytometry. The concentrations of IL-6 and IL-12 in the supematant were measured by enzyme-linked immunosorbent assay （ELISA）. T cell proliferation induced by DC was detected by methyl thiazolyl tetrazolium （MTT）. RESULTS： The cells treated with 0.5 mmol/L lamivudine had the highest expression of CDla, CD83 and HLA-DR, and the expression of CD80 was not significantly different between the cells with and without lamivudine treatment. After treatment with 0.5 mmol/L lamivudine, the expression of CDla, CD83 and HLA-DR were significantly higher than those of DCs without lamivudine treatment （CDla： 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）; the secretion of IL-12 was significantly increased in comparison with that of the control group （810.0 ± 91.5 ng/L vs 268.0 ± 34.3 ng/L, P 〈 0.05）, while the level of IL-6 was lowered markedly （28.1 ± 2.6 ng/L vs 55.3 ± 7.4 ng/L, P 〈 0. 05）; the stimulatory capacity of DCs in the allogeneic mixed leukocyte reaction was markedly enhanced as compared with that of the control group （stimulatory index： 1.9 ± 0.6 vs 1.2 ± 0.5, P 〈 0.05）. CONCLUSION： Pulsed with lamivudine in certain concentration, the biological activity of DCs derived from CHB patients can be efficiently enhanced.