摘要
目的 建立西尼罗病毒(West Nile virus,WNV)假病毒的体内感染模型,并对抗体WNV-XH1进行体内中和活性评价.方法 利用前期建立的可包装WNV假病毒的稳定细胞株制备假病毒上清,将上清浓缩后感染BHK21细胞检测假病毒滴度.假病毒经腹腔注入C57BL/J小鼠体内后进行生物发光成像,观测小鼠体内假病毒感染情况.同时感染后取血,ELISA检测小鼠血清中NS1水平.利用建立的小鼠感染模型,评价抗体WNV-XH 1的体内功能活性.结果 C57BL/J小鼠感染WNV假病毒后,体内可检测到荧光,且感染假病毒的小鼠与未感染的小鼠相比外周血血清中WNV非结构蛋白NS1水平明显升高(1.453±0.09vs0.305±0.018).攻毒前静脉给予抗体WNV-XH 1后,小鼠体内荧光信号减弱,血清中NS1水平下降(0.384±0.015).结论 成功建立WNV假病毒体内感染模型,并证实抗体WNV-XH 1在体内对WNV假病毒的感染具有保护作用.
Abstract
Objective To establish an in vivo infection model of West Nile virus(WNV)pseudovirus and evaluate the neutralizing activity of antibody WNV-XH1.Methods A stable cell line that can package the WNV pseudovirus was established in the early stage to prepare the pseudovirus supernatant.The supernatant was concentrated and infected BHK21 cells to detect the titer of the pseudovirus.After intraperitoneal injection of the pseudovirus into C57BL/J mice,bioluminescence imaging was performed to observe the infection status of the pseudovirus in the mice.After simultaneous infection,blood was collected and ELISA was used to detect NS1 levels in mouse serum.The in vivo functional activity of antibody WNV-XH1 was evaluated using the established mouse infection model.Results Fluorescence was detected in C57BL/J mice infected with WNV pseudovirus,and the NS1 levels in the peripheral blood serum of mice infected with pseudovirus were significantly higher than those of non infected mice(1.453±0.09vs0.305± 0.018).After intravenous administration of WNV-XH1 antibody before the attack,the fluorescence signal in the mice decreased and the serum NS1 level decreased(0.384±0.015).Conclusions A successful in vivo infection model of WNV pseudovirus was established,and it was confirmed that the antibody WNV-XH1 had a protective effect against WNV pseudovirus infection in vivo.
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