整合狂犬病病毒G蛋白的假病毒构建及应用

Construction and application of pseudovirus integrating rabies virus G protein

目的 构建表达狂犬病病毒(rabies virus, RABV)G蛋白的假病毒,为RABV中和抗体检测及疫苗的研发提供有效实验工具。方法 通过无缝克隆技术构建表达RABV G蛋白的假病毒p-RABV-G,通过抗体中和试验评估狂犬疫苗接种者体内的中和抗体水平。结果 成功在293T细胞内进行假病毒p-RABV-G的包装,p-RABV-G可以表达G蛋白,并能高效感染Huh7.5细胞。4位狂犬疫苗接种者21 d的血清对p-RABV-G的抑制效果处于最优水平,在1∶625倍数的稀释条件下对p-RABV-G的抑制率高达95.79%、98.26%、98.42%和98.88%。结论 成功构建表达RABV G蛋白的假病毒p-RABV-G可用于狂犬疫苗的开发与中和抗体的筛选。在狂犬疫苗接种后的111 d,本文调查的4位接种者中的其中3位体内血清仍可对p-RABV-G产生抑制效果。

Objective To construct a pseudovirus expressing RABV G proteinand provide an effective experimental tool for the detection of RABV neutralizing antibody and the development of vaccine. Methods Seamless cloning technology was used to construct the pseudovirus p-RABV-G which expressed RABV G protein, and antibody neutralization assay was operated to evaluate the neutralizing antibody level of vaccine recipients. Results Pseudovirus p-RABV-G was packaged in 293T cells, p-RABV-G could express G protein and efficiently infect Huh7.5 cells. The inhibition of p-RABV-G by the serum of 4 rabies vaccine recipients on day 21 was at the optimal level, and the inhibition rates of p-RABV-G were as high as 95.79%, 98.26%, 98.42% and 98.88% under the dilution condition of 1∶625. Conclusion The constructed pseudovirus p-RABV-G expressing RSBV G protein in this study could be used in the development of rabies vaccine and the screening of neutralizing antibodies. 111 days after vaccination, 3 serum of of the 4 recipients still inhibited p-RABV-G.