论文部分内容阅读
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。
OBJECTIVE: To express the recombinant adenovirus (AdenoG2) of Hantaan virus (HTNV) glycoprotein G2 in VeroE6 cells and to investigate its immunological response. Methods: Vero E6 cells were infected with HTNVAdenoG2 virus (titer of about 1 × 1013 pfu / L) and their expression products were detected by IFA. After the BALB / c mice were immunized with the expressed product, the humoral and cellular immune responses were detected by ELISA, micro-cell culture neutralization assay and lymphocyte proliferation assay. Results: The expression of HTNV glycoprotein G2 was detected after infection of VeroE6 cells with HTNVAdenoG2. After immunization of mice with HTNVAdenoG2, a specific antibody against HTNV glycoprotein G2 can be induced with an antibody titer of 1:40. The results of micro-cell culture neutralization assay showed that HTNVAdenoG2 can stimulate mice to produce low levels of neutralizing antibodies; however, lymphocyte proliferation reaction was not obvious. Conclusion: HTNV glycoprotein G2 was successfully expressed in VeroE6 cells. After the mice were immunized with HTNVAdenoG2, the mice mainly stimulated to produce specific anti-HTNV humoral immune response, but the specific cellular immune response was not obvious. The results of this study provide experimental evidence for the development of HTNV genetic engineering vaccine.