体外合成JSRV mRNA及其体内表达特异性中和抗体的研究

Synthesis of JSRV mRNA in vitro and its expression of specific antibodies in mouse

目的:应用假病毒中和法检测体外合成JSRV-env mRNA免疫小鼠产生的特异性抗体。方法:优化PT7TS载体序列,插入目的序列经电泳测序验证后,体外转录合成JSRV-env mRNA,将mRNA与鱼精蛋白结合后皮下注射免疫BALB/c小鼠。采用慢病毒包装系统制作JSRV假病毒,测定假病毒滴度,取不同组免疫小鼠血清进行假病毒为基础的抗体中和实验,检测小鼠体内针对JSRV-env mRNA的特异性抗体。结果:成功体外合成具有稳定性的JSRV-env mRNA;重组了基于慢病毒质粒结构的JSRV假病毒,且具有感染能力;假病毒体外中和实验检测出JSRV-env mRNA免疫小鼠体内产生了针对JSRV的特异性抗体。结论:针对JSRV病毒包膜的信使RNA,经皮下注射到小鼠体内,能够表达产生针对JSRV-env的中和抗体。

Objective： To detect the specific antibodies of JSRV-env mRNA immunized mice by pseudovirus neutralization method.Methods： The PT7 TS vector was optimized before inserted in the target sequence. The recombinant plasmid of PT7TS-JSRV-env was verified by electrophoretic. After verifying its correctin in vitro,the JSRV-env mRNA was synthesized. The BALB/c mice were given subcutaneous injection with protamine and JSRV-env mRNA complex. The specific antibodies of JSRV-env mRNA immunized mice was determined by the pseudovirus neutralizing antibody test. The JSRV pseudovirus was produced by the lentivirus packaging system. Results：The JSRV-env mRNA was successfully synthesized in vitro. The JSRV pseudovirus was recombined and presented powerful infection ability. The neutralization test of the sample serum in immunized mice and JSRV pseudovirus verified the JSRV pseudovirus. Conclusion：JSRV-env producing antibodies can be expressed in mice by subcutaneous injections into JSRV-env eloped messenger RNA.