丙型肝炎病毒特异性细胞毒性T淋巴细胞功能的研究  

Study of the function of HCV-specific cytotoxic T lymphocytes in chronic hepatitis C

作　　者：孙丽杰[1] 于建武[1] 李树臣[1] 李明荣[1] 

机构地区：[1]哈尔滨医科大学附属第二医院感染病科,150086

出　　处：《肝脏》2007年第2期87-90,共4页Chinese Hepatology

摘　　要：目的研究慢性丙型肝炎患者丙型肝炎病毒(HCV)特异性细胞毒性T淋巴细胞(CTL)的功能及其与临床疾病状态的关系。方法表达HCV核心蛋白的真核表达质粒pcDNA3.1-core通过Lipofecta-mineTM基因转染法转染HepG2细胞,经G418筛选获得稳定转染HepG2细胞(Hep-core),经Western blot证实有HCV核心蛋白表达;分离患者外周血单个核细胞,经体外诱导扩增获得HCV特异性CTL(HCV-CTL),以Hep-Core细胞和HepG2细胞作靶细胞,乳酸脱氢酶释放法检测HCV-CTL活性,用酶联免疫吸附法检测培养上清液中干扰素-γ(IFN-γ)含量,以正常人作为对照。结果慢性丙型肝炎患者组HCV-CTL活性值为(23.9±4.8)%,明显低于对照组(42.6±6.5)%(t=7.22,P=0.011)。高病毒载量组HCV-CTL活性值(18.9±4.8)%,明显低于低病毒载量组的(33.7±3.2)%(t=8.22,P=0.003);基因1型患者HCV-CTL活性值(20.8±2.1)%明显低于基因2或3型的(32.4±2.5)%(t=11.7,P=0.001);ALT升高患者HCV-CTL活性值(29.3±3.1)%与ALT正常患者(25.7±3.4)%相比无统计学差异(t=0.93,P>0.05)。慢性丙型肝炎患者培养上清液中的IFN-γ量(957±241)pg/ml明显低于对照组的(3117±673)pg/ml(t=8.87,P=0.001)。结论慢性丙型肝炎患者HCV-CTL活性降低,CTL的免疫功能低下与病毒水平和基因型相关而与ALT水平无关。Objective To study the function of HCV-specific cytotoxic T lymphocytes （HCV-CTL） and its influencing factors in patients with chronic hepatitis C. Methods A cell line expressing stably HCV core protein-HepG2-core was constructed by gene transfecting the plasmid pcDNA3.1-core （expressing HCV core protein） into HepG2 by Lipofecta- mine^TM . HepG2 cells transfected with recombinant eukaryotic expression plasmid were selected by G418. The HCV-C protein in HepG2 cells transfected with recombinant plasmid was verified by Western blot assay. HCV-CTL got from PBMC and stimulated by HCV antigen peptide, activity of HCV-CTL were tested by LDH releasing assay, the levels of IFN-γ of cells culture were detected by ELISA assay. 22 HLA-A2 positive patients with chronic hepatitis C were tested and 8 healthy subjects as controls. Results Activity （23.9 ± 4.8） % of HCV-specific CTL of patients with chronic hepatitis C was significant lower than that （42.6 ± 6.5） % of the normal subjects （ t = 7.22, P = 0.011 ）. Activity （ 18.9 ± 4.8） % of HCV-spocific CTL of patients with high viral load was significant lower than that （33.7 ± 3.2） % of the patients with low viral load （ t = 8,22, P = 0.003）, Activity （20.8 ± 2.1 ） % of HCV-specific CTL of patients with genotype 1 was significant lower than that （32.4±2.5）% of the patients with genotype 2 or 3（t = 11.7,P=0.001）. Activity （29.3± 3.1）% of HCV-specific CTL of patients with high levels of serum ALT was no significant difference with that （25.7 ± 3.4） % of the patients with normal levels of ALT （t = 0.93, P 〉 0.05）. The concentration （957 ± 241 ）pg/ml of IFN-γ of cells culture of patients with chronic hepatitis C was significant lower than that （3 117 ± 673）pg/ml of controls （ t = 8.87, P = 0.001）. Conclusion The activity of HCV-CTL of patients with chronic hepatitis C was low, which was related with concentration of HCV-RNA and genotype, but not the levels of ALT.

关 键 词：丙型肝炎病毒 特异性细胞毒性T淋巴细胞 干扰素-Γ 

分 类 号：R512.63[医药卫生—内科学]