HBsAg联合rhIL-12对慢性乙型肝炎患者PBMC诱生IFN-γ的作用  

Combination of HBsAg and rhIL-12 induces the production of IFN-γ of PBMC from chronic hepatitis B patients in vitro

作　　者：王翠玲 李茹冰[2] 王增松 夏书奇 段祥 谢飞群 曾振飞 揭燕 傅泳航[2] 张宜俊 

机构地区：[1]广州市恺泰生物科技有限公司,广东广州510620 [2]解放军第458医院检验科,广东广州510602

出　　处：《检验医学》2009年第9期671-675,共5页Laboratory Medicine

摘　　要：目的确认重组人白细胞介素12(rhIL-12)联合乙型肝炎表面抗原(HBsAg)可增强慢性乙型肝炎患者外周血单个核细胞(PBMC)对HBsAg产生特异性细胞免疫应答,探讨rhIL-12联合HBsAg作为治疗型乙型肝炎疫苗的应用前景。方法取8名正常人和16例慢性乙型肝炎患者的PBMC,分别用不同浓度的HBsAg、rhIL-12单独体外刺激,再用HBsAg联合rhIL-12(0.01、0.1和1.0ng/mL)刺激患者PBMC,72h后用酶联免疫吸附试验(ELISA)测定培养上清中的γ干扰素(IFN-γ)。结果HBsAg或rhIL-12单独刺激仅产生低水平的IFN-γ,联合使用HBsAg和rhIL-12刺激后,患者PBMC产生的IFN-γ平均水平显著高于单独使用HBsAg或rhIL-12(P均<0.05)。HBsAg(0.5μg/mL)联合rhIL-12(0.01、0.1和1.0ng/mL)刺激患者PBMC产生的IFN-γ与rhIL-12的浓度之间呈明显的量效关系。结论HBsAg和rhIL-12联合刺激乙型肝炎患者PBMC,通过产生高水平IFN-γ激活乙型肝炎患者特异性细胞免疫。Objective To confirm the enhancing effect of recombinant human interleukin 12 （rhIL-12）-hepatitis B surface antigen （HBsAg） combination on the cellular immune response to HBsAg of peripheral blood mononuclear cell （PBMC） from chronic hepatitis B patients in vitro. To investigate the foreground of HBsAg-rhIL-12 combination being a novel therapeutic hepatitis B vaccine. Methods PBMC were prepared from the peripheral blood of 8 healthy subjects and 16 chronic hepatitis B patients and cultured in the presence of variant concentration HBsAg or rhIL-12. The PBMC were cultured in the presence of variant concentration HBsAg and rhIL-12（0.01,0.1 and 1.0 ng/mL）. The level of interferon-γ（IFN-γ） in culture supernatants which harvested at 72 h were detected by enzyme-linked immunosorbent assay （ELISA）. Results Low levels of IFN-γwere observed when PBMC from healthy subjects and chronic hepatitis B patients were stimulated with HBsAg or rhIL-12 alone. The IFN-γ levels in culture supernatants of PBMC from chronic hep- atitis B patients with HBsAg-rhIL-12 combination increased significantly（P 〈 0. 05 ）. When HBsAg（0.5μg/mL）plus rhIL-12（0.01,0.1 and 1.0 ng/mL）stimulated the PBMC from chronic hepatitis B patients,the concentration of rhIL-12 was with the dose-dependant to produce IFN-γ. Conclusions rhIL-12 combined with HBsAg can promote the specific cellular immune response to the Hepatitis B virus by the enhancement of IFN-γproduction.

关 键 词：乙型肝炎表面抗原 重组人白细胞介素12 外周血单个核细胞 细胞免疫 Γ干扰素 

分 类 号：R446.62[医药卫生—诊断学]