猪可溶性CD40L三聚体蛋白的原核表达及其对口蹄疫病毒样颗粒疫苗免疫效果评价

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中图分类号：S855 文献标志码：A 文章编号：0366-6964（2025）10-5072-12

Abstract： This study aimed to evaluate the immune-enhancing effect of porcine soluble CD4OL trimer protein on the Virus-like particles（VLPs） vaccine of Foot-and-mouth disease （FMD）. In this study，the plasmid containing porcine CD40 ligand （CD40L） trimer（5B CD40L） gene was expressed in Escherichia coliBL21（DE3）. The purified recombinant 5B CD4OL protein was coupled to FITC and reacted with PBMCs. The biological activity of CD4OL protein reacting with monocytes was studied by fluorescence microscopy. The guinea pigs were further immunized with 5B CD4OL，VLPs and 5B CD40L-VLPs，and the effects of 5B CD40L on the immune effect of VLPs vaccne were evaluated by neutralizing antibodies，splenic lymphocyte proliferation and real-time RT-qPCR. SDS-PAGE and Western blot results showed that the size of recombinant 5B CD40L protein expressed was about ，The optimal expression conditions were 16∘C OD600nm （20 value was O. 9，and concentration was 0.5mmolL-1 after IPTG induction for 12h . Flow cytometry analysis showed that the recombinant CD40L protein could bind to CD4O protein molecules on the surface of porcine peripheral blood mononuclear cells （PBMCs）. The combination of 5B CD4OL and VLPs can accelerate the reaction of specific antibody and neutralizing antibody，and improve the proliferation level of spleen lymphocytes. In the 5B CD40L-VLPs group，the IL-2 and INF-γ mRNA of spleen cells are significantly higher than those in the VLPs group. In summary，5B CD4OL has the ability to enhance humoral and cellular immune responses of FMD VLPs vaccine，and has the potential as a vaccine adjuvant.

Keywords：5B CD4oL;Foot-and-mouth disease;immune effect ∗ Corresponding authors： MU Suyu，E-mail： mutou6633937 126.com;SUN Shiqi，E-mail：sun shiqi@ caas. cn

CD40L，又称CD154或gp39，由于其结构与肿瘤坏死因子（tumornecrosisfactor，TNF）基因超家族其他成员的高度同源性，被归类为TNF超家族成员，是一种II型跨膜蛋白[1]。（剩余19733字）