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目的研究重组表达的新疆出血热病毒(XHFV)核蛋白和糖蛋白截短片段在刺激机体免疫应答中的作用。方法采用基因重组技术和亲和层析技术对XHFV截短的核蛋白(NP2)、糖蛋白片段(Gc和Gn)进行表达和纯化,并通过蛋白免疫及核酸初次免疫、蛋白加强免疫的方式免疫BALB/c小鼠,通过ELISA检测鼠血清抗体效价,采用T淋巴细胞增殖试验检测细胞免疫应答的效果,对上述表达产物刺激小鼠免疫应答的作用进行初步评价。结果利用原核表达系统成功表达纯化出XHFV核蛋白和糖蛋白截短片段。重组蛋白免疫BALB/c小鼠后能刺激机体产生Ig G(效价为1∶12 800以上),并能与相应的抗XHFV核蛋白及糖蛋白的多克隆抗体发生特异性反应。各组均能有效地激发较强的细胞免疫应答,其中p VAX1-Gc联合r Gc实验组小鼠脾脏淋巴细胞增殖明显,与对照组相比,差异具有统计学意义(P<0.01)。结论获得的重组XHFV核蛋白、糖蛋白片段能有效刺激小鼠的体液和细胞免疫应答。
Objective To investigate the role of recombinant XHFV nucleoprotein and glycoprotein truncated fragments in stimulating the immune response in vivo. Methods Recombinant protein and affinity chromatography were used to express and purify the truncated nucleoprotein (NP2) and glycoprotein fragments (Gc and Gn) of XHFV. BALB / c mice were immunized with protein immunization and primary immunization / c mice. Serum antibody titers were measured by ELISA. The effect of cellular immune response was detected by T lymphocyte proliferation test. The effects of the above expression products on the immune response in mice were preliminarily evaluated. Results The recombinant protein of XHFV and glycoprotein truncated fragments were successfully expressed and purified by using prokaryotic expression system. The recombinant protein can stimulate the body to produce Ig G (titer of 1:12 800) after immunizing BALB / c mice and can specifically react with the corresponding polyclonal antibodies against XHFV nucleoprotein and glycoprotein. Each group can effectively stimulate a strong cellular immune response, in which p VAX1-Gc combined with r Gc mice spleen lymphocyte proliferation significantly, compared with the control group, the difference was statistically significant (P <0.01). Conclusion The obtained recombinant XHFV nucleoprotein and glycoprotein fragments can effectively stimulate humoral and cellular immune responses in mice.