Hsp70与汉滩病毒S基因融合蛋白的原核表达及免疫小鼠的初步研究

本文构建了hsp70与S基因的原核融合表达载体pGEX-4T-1/hsp70-S,在大肠杆菌中表达,并通过GSTrapFF柱进行了纯化。同时制备了NP和Hsp70两种纯化蛋白。分别用这三种纯化蛋白免疫BALB/c小鼠,结果表明纯化的NP和Hsp70-NP两种蛋白均可同时诱导产生抗汉滩病毒核蛋白(NP)抗体,且后者刺激产生的抗体效价明显高于前者。淋巴细胞增殖实验表明,两组免疫小鼠的脾细胞均能够对体外抗原刺激产生增殖反应,而Hsp70-NP组免疫小鼠脾细胞对NP的增殖指数明显高于NP组免疫组。结果显示,与单独用NP免疫小鼠相比,Hsp70-NP纯化蛋白可以刺激机体产生更强的抗汉滩病毒体液免疫应答和特异性淋巴细胞增殖反应。

Prokaryotic Expression and Purification of Hsp70-NP and Its Immunity

Recombinant fusion prokaryotic expression vector pGEX-4T-1/hsp70-S was constructed by cloning genes Hsp70 and Hantaan virus S gene coding region into pGEX-4T and was expressed in E.coli.The Hsp70-NP fusion protein was separated and purified with GSTrapFF purification system.We also constructed the prokaryotic expression vectors pGEX-4T-1/hsp70 and pET28a/S to prepare the purified protein Hsp70 and NP.Then BANB/c mice were vaccinated with the protein Hsp70-NP,Hsp70 and NP separately.ELISA results showed that both Hsp70-Npand NP could induce specific antibodies against NP.The specific antibody titers stimulated by Hsp70-NP were obviously higher than that of NP.The lymphoproliferative responses of spleen cells clearly showed that the spleen cells from both groups of mice were able to proliferate in the presence of NP protein.The stimulation indexes of splenocytes to NP induced by Hsp70-NP were significantly higher than that induced by NP.It indicated that linkage of Hsp70 to NP enhanced the immune response to Hantaan virus nucleocapsid protein.