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目的从马尔堡病毒GP蛋白上寻找与GP免疫原性相当的小片段用于疫苗设计,以克服全长蛋白缺陷。方法基于GP(aa:1~681)结构及功能分区,构建3个片段[GP1△(aa:25~239)、GPM(aa:250~520)、GP2△(aa:436~648)],经原核表达后免疫小鼠,不同时间检测血清特异抗体滴度,检测脾淋巴细胞增殖指数及测定细胞因子浓度。结果 ELISA检测结果显示,GP2△组与GP混合组(GP1△+GPM+GP2△)产生体液免疫能力相当,明显高于GP1△与GPM组(P<0.05);Con A非特异刺激后,GP2△组脾淋巴细胞的SI值高于GP混合组(P<0.05),GP混合组特异刺激后,GP2△组脾淋巴细胞的刺激指数(SI)值低于GP混合组(P<0.05);细胞因子检测中,GP2△组的白细胞介素2(IL-2)及干扰素γ(IFN-γ)浓度均高于对照组,低于GP混合组(P<0.05)。结论马尔堡病毒GP2△片段免疫小鼠能诱导与完整GP蛋白相当的体液免疫,同时也能诱导一定的细胞免疫,可用来替代完整GP用于疫苗设计。
OBJECTIVE: To search for a small fragment of immunogenicity equivalent to GP from Marburg virus GP protein for vaccine design to overcome full-length protein defects. Methods Three fragments (GP1 △ (aa: 25 ~ 239), GPM (aa: 250 ~ 520) and GP2 △ (aa: 436 ~ 648) were constructed based on the structure and functional partition of GP (aa: 1-681) After the prokaryotic expression, the mice were immunized, the serum specific antibody titers were detected at different times, the splenic lymphocyte proliferation index and cytokines concentration were measured. Results The results of ELISA showed that the humoral immunity of GP2 △ group and GP mixed group was quite higher than that of GP1 △ and GPM group (P <0.05). After Con A non-specific stimulation, GP2 The SI value of splenic lymphocytes in △ group was higher than that in GP group (P <0.05). The spleen lymphocyte stimulating index (SI) of GP2 △ group was lower than that of GP group (P <0.05) after stimulated by GP. In cytokines, the concentrations of IL-2 and IFN-γ in GP2 △ group were higher than those in control group and GP group (P <0.05). Conclusion The Marburg virus GP2 △ fragment immunized mice can induce humoral immunity equivalent to the intact GP protein and induce certain cellular immunity, which can be used to replace the complete GP for vaccine design.