香菇多糖联合丙肝核酸疫苗诱导抗原特异性免疫应答研究  

Antigen-specific immunity generated by lentinan plus HCV NS3 DNA vaccine

作　　者：邹镇[1] 彭佳蓓[1] 谢潜红[1] 向明[1] 

机构地区：[1]华中科技大学同济药学院药理室,武汉430030

出　　处：《中国免疫学杂志》2008年第4期348-351,共4页Chinese Journal of Immunology

基　　金：湖北省卫生厅科研基金项目(编号:NX200601)

摘　　要：目的:探讨香菇多糖(Lentinan,LNT)联合丙型肝炎病毒(HCV)非结构蛋白NS3核酸疫苗诱导NS3抗原特异性免疫作用。方法:将不同剂量香菇多糖与等量HCVNS3质粒(100μg/只),采用初次加强免疫法联合免疫BALB/c小鼠。第二次免疫10天后取小鼠脾脏,制备脾细胞悬液,与NS3抗原多肽共育作为效应细胞。另培养NS3抗原致敏的P815细胞作为靶细胞。采用MTT法分别检测NS3抗原特异性T细胞增殖反应和细胞毒T细胞特异性杀伤反应(CTL)。同时采用ELISA方法检测血清中细胞因子IL-4和IFN-γ分泌水平。结果:研究发现高剂量及中剂量香菇多糖与NS3质粒联合在效靶细胞比为50:1时,可明显加强NS3抗原特异性CTL反应,与单用NS3质粒相比有显著差异(P<0.01和P<0.05);中剂量和低剂量香菇多糖与NS3质粒联用明显增强NS3抗原特异性T细胞增殖反应(均为P<0.01);细胞因子分析显示中剂量和低剂量香菇多糖与NS3质粒联用使IFN-γ分泌增加(均为P<0.01)。结论:采用初次加强免疫法将中剂量香菇多糖(0.1ml/10g)与HCVNS3质粒联合免疫BALB/c小鼠,可激发明显增强的抗原特异性T细胞增殖和CTL反应,并伴有大量的IFN-γ分泌,提示香菇多糖可作为丙肝病毒NS3核酸疫苗的免疫佐剂,加强抗原特异性免疫应答效应。Objective：To investigate the effects of lentinan combined with HCV NS3 DNA vaccine on the antigen specific immune response.Methods：BALB/c mice were ip with different dose of lentinan plus the same dose of NS3 plasmid（100 μg/per mouse） in a prime and boost immunization strategy.10 days after the second injection,spleens were taken.The cell suspension was prepared and cultured with NS3 peptide as the stimulant.In addition,NS3 cocultured P815 cells were used as target cells.NS3 specific T cell proliferation and cytotoxicity T lymphocyte reaction（CTL）were analyzed by MTT methods,and the cytokines IL-4 and IFN-γ in serum were detected by ELISA technique.Results：It was showed that the high and middle dose of lentinan plus NS3 plasmid specifically stimulated NS3 specific CTL reaction at the ratio of effector to target as 50∶1,compared with the single use of NS3 plasmid,P〈0.01 and P〈0.05,respectively.T cell proliferation was significantly promoted in the middle dose and low dose of lentinan plus NS3 plasmid groups（both were P〈0.01）.Cytokine analysis showed that IFN-γ secretion were increased in both middle dose and low dose of lentinan plus with NS3 plasmid groups as well（both were P〈0.01）.Conclusion：Immunization of BALB/c mice with HCV NS3 plasmid plus the middle dose of lentinan（0.1 ml/10 g） could induce strong antigen specific CTL reaction and T cell proliferation,accompanying with increased IFN-γ secretion.It is suggested that lentinan could be used as adjuvant for HCV NS3 vaccine to provoke antigen specific immune response.

关 键 词：丙型肝炎 非结构蛋白3 香菇多糖 核酸疫苗 

分 类 号：R392[医药卫生—免疫学]