马流产沙门菌重组鞭毛蛋白FliC和FljB免疫原性比较

Comparison of immunogenicity between recombinant flagellins C and B of Salmonella abortus equi

摘要:

为评价与比较马流产沙门菌 (Salmonella abortus equi) 2种不同鞭毛蛋白FliC (Flagellin C) 和FljB (Flagellin B) 的免疫原性，为进一步利用这两种蛋白奠定实验基础，本研究诱导表达及纯化FliC和FljB蛋白，将纯化后的蛋白分别免疫小鼠，对免疫后小鼠的抗体水平、滴度及抗体亚型进行检测，攻击小鼠后进行免疫相关受体检测及病理组织学观察。结果显示，成功诱导表达了重组蛋白FliC和FljB，纯化后蛋白的相对分子量分别为52 kDa和42 kDa。两种蛋白分别免疫小鼠，产生较高水平的特异性抗体IgG，FljB免疫组抗体水平高于FliC免疫组，且抗体亚型以IgG1为主。FljB免疫组攻击保护率为87.5%，高于FliC免疫组的75%，病理组织学及脏器荷菌数检测结果显示，FljB免疫组效果优于FliC免疫组；FljB免疫组诱导TLR2、TLR4、MHC-I、TCR (T细胞抗原受体) 的水平均高于FliC免疫组。该结果表明，FljB免疫组诱导小鼠免疫应答的水平高于FliC免疫组。

Abstract:

To evaluate and compare of the immunogenicity differences of flagellins FliC and FljB of Salmonella abortus equi, and lay the experimental foundation for the further utilization of the two recombinant proteins, FliC and FljB recombinant proteins were induced, expressed and purified. The purified FliC and FljB were used to immunize mice separately. The antibody level, titer and subtype of mice serum were detected after immunization. Immune-related receptors and histopathological changes were observed in immunized mice after challenged. The recombinant proteins FliC and FljB were successfully induced and expressed. Proteins of about 52 kDa and 42 kDa were purified. High levels of specific IgG antibodies were induced in mice immunized with these two proteins, the antibody level of FljB-immunized group was higher than that of FliC-immunized group, and IgG1 was the dominant subtype of antibody. The challenge protection rate of the FljB-immunized group was 87.5%, higher than that of FliC immunized group. Bacterial loads and observation pathological of FljB-immunized group were better than that of FliC immunized group, the levels of TCR2, TCR4, MHC-I and TCR induced by FljB-immunized group were higher than those of FliC-immunized group. The of immune response induced by FljB group was better than that of FliC group.