汉滩病毒G2重组腺病毒的表达及其基因免疫的研究

目的:在VeroE6细胞中表达汉滩病毒(HTNV)囊膜糖蛋白G2重组腺病毒(AdenoG2),并探讨其诱导免疫应答特性。方法:以HTNVAdenoG2病毒原种(滴度约1×1013pfu/L)感染VeroE6细胞,用IFA法检测其表达产物。以表达产物免疫BALB/c小鼠后,用ELISA、微量细胞培养中和试验及淋巴细胞增殖试验检测体液及细胞免疫应答。结果:用HTNVAdenoG2感染VeroE6细胞后,可检测到HTNV糖蛋白G2的表达。用HTNVAdenoG2免疫小鼠后,可诱导产生抗HTNV糖蛋白G2的特异性抗体,抗体效价为1∶40。微量细胞培养中和试验的结果表明,HTNVAdenoG2还可刺激小鼠产生低水平的中和抗体;但淋巴细胞的增殖反应不明显。结论:在VeroE6细胞中成功地表达了HTNV糖蛋白G2。以HTNVAdenoG2免疫小鼠后,主要刺激小鼠产生特异性的抗HTNV体液免疫应答,而特异性的细胞免疫应答不明显。本研究结果为HTNV基因工程疫苗的研制提供了实验依据。

Expression and genetic immunization of hantaan virus G2 recombinant adenovirus

AIM: To express hantaan virus(HTNV) envelope glycoprotein G(2) recombinant adenovirus(Adeno-G(2)) in vero E6 cells and explore its property of inducing immune response. METHODS: Vero E6 cells were infected with the HTNV Adeno-G(2) (100 MOI). The expression of Adeno-G(2) in the infected Vero E6 cells was detected by IFA. BALB/c mice were immunized with HTNV Adeno-G(2), then the immune response to Adeno-G(2) was tested by ELISA, microcell-culture neutralizing experiment and lymphocyte proliferation test (MTT colorimetry). RESULTS: IFA detection showed the expression of Adeno-G(2) in the infected Vero E6 cells. The titer of specific antibody was 1:40; The low-titer neutralization antibody was also detected. But the lymphocyte proliferation reaction was not notable. CONCLUSION: The HTNV Adeno-G(2) can stimulate BALB/c mice to develop specific humoral immune response instead of specific cell-mediated immunity. This study provides the experimental basis for the development of gene engineering vaccine of HFRS.