目的探讨原发性胆汁性肝硬化(primary biliary cirrhosis,PBC)患者共刺激因子B7-H4的表达及其与疾病发病机制的关系。方法分别采用荧光实时定量PCR法(real-time PCR)、酶联免疫吸附试验(ELISA)及流式细胞术(FCM)检测65名PBC患者外周血单个核细胞(PBMC)B7-H4 mRNA表达水平、血清IL-2水平以及CD4^+、CD8^+T细胞亚群和T细胞表面B7-H4表达百分率,同时监测抗线粒体抗体(anti-mitochondrial antibody,AMA)及临床各项生化指标的关系。结果(1)PBC患者PBMC B7-H4 mRNA水平及T细胞表面B7-H4表达百分率显著低于非PBC肝硬化组及健康对照组(P〈0.01)。(2)活化72h后各实验组及对照组IL-2含量及CD4^+、CD8^+、CD4^+CD8^+T淋巴细胞表达水平均低于活化前,以非PBC肝硬化组与健康对照组降低显著(P〈0.05);PBC组IL-2含量及CD4^+、CD4^+CD8^+T淋巴细胞表达水平高于非PBC肝硬化组与健康对照组(P〈0.01)。(3)AMA-M2阳性患者血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、碱性磷酸酶(ALP)及γ-谷氨酰转肽酶(GGT)水平升高,其中ALP及GGT升高显著(P〈0.05);AMA—M2阳性患者与阴性患者T细胞表面B7-H4表达百分率差异无统计学意义(P〉0.05)。结论共刺激因子B7-H4对PBC患者体内T细胞活化增殖及细胞因子分泌的抑制作用减弱,为研究PBC的发生发展和阐明PBC的发病机制提供了试验资料。
Objective To study the relationship between the expression of costimulating factor BT- H4 in patients with primary biliary cirrhosis (PBC) and the pathogenesis of PBC. Methods The expression of B7-H4 mRNA on peripheral blood mononuclear cell (PBMC) of 65 patients with PBC was tested by realtime PCR. Serum levels of IL-2 were assayed by ELISA. CD4^ + , CD8^ + T lymphocytes expression level and B7-H4 expression rate before and after activation were measured by three-color flow cytometry (FCM). Resuits ( 1 ) Expression of B7-H4 mRNA and BT-H4 percentage in PBC group were significantly lower than that in none-PBC group and healthy controls ( P 〈 0.01 ) ; ( 2 ) After 72 h activation, the percentage of CD4^+ , CD8 ^+ , and CD4 ^+ CD8 ^+ T lymphocytes and serum levels of IL-2 decreased ( P 〈 0.05 ), and the percentage of CD4 ^+ , CD4 ^+ CD8 ^+ T lymphocytes and serum levels of IL-2 were significantly higher than that of none- PBC group and healthy controls (P 〈 0.01 ) ;( 3 ) Levels of alanine aminotransferase (ALT), aspartate aminotransferase( AST), alkaline phosphatase(ALP) and γ-glutamyl transpeptidase(GGT) of patients with posi- tive anti-mitochondrial antibody(AMA)-M2 rose. There was no significant difference of BT-H4 expressions on T cells between patients either with or without AMA-M2 antibody. Conclusion The costimulating factor BT-H4 can express on T lymphocytes which is activated by phytohemagglutinin(PHA) and plays a negative role on T cells responses.