摘要： 为了研究坦布苏病毒囊膜蛋白（E蛋白）的免疫原性，利用大肠杆菌表达系统高效表达坦布苏病毒囊膜蛋白（E蛋白）并对其诱导小鼠免疫应答进行了研究。结果表明，E蛋白免疫BALB/c小鼠后，免疫组小鼠血清中抗体效价均可达1∶51 200以上；同时，E蛋白可显著诱导血清中细胞因子IL-2、TNF-α和IFN-γ的表达，免疫组小鼠血清中IL-2、TNF-α的含量均极显著高于对照组（P<0.01），IFN-γ含量则显著高于对照组（P<0.05）；此外，与对照组相比，E蛋白还可极显著刺激小鼠脾脏淋巴细胞的增殖，上述结果表明，E蛋白免疫后，小鼠机体可产生显著的体液免疫和细胞免疫反应。攻毒保护性试验结果显示，攻毒后，经荧光定量RT-PCR检测，E蛋白免疫组小鼠脑、肝脏、脾脏、卵巢、肾脏中坦布苏病毒载量均显著低于对照组，表明免疫E蛋白可有效抑制坦布苏病毒在小鼠各个组织中的增殖。结果表明，大肠杆菌表达的坦布苏病毒E蛋白可作为坦布苏病毒亚单位疫苗的候选抗原，为进一步预防和控制坦布苏病毒病奠定理论基础。

关键词: 坦布苏病毒, 囊膜蛋白, 免疫应答, 亚单位疫苗

Abstract: In order to research the immunogenicity of Goose tembusu virus (GTV) envelope protein (E protein),GTV E protein was expressed in Escherichia coli, and administered to BALB/c mice via subcutaneous injection to study the immune responses. After immunization by recombinant E protein, the titres of neutralizing antibody in immunized BALB/c mice were all above 1:51 200. Cytokine analysis of the serum of mice immunized with recombinant E protein revealed antigen-specific IL-2, TNF-α and IFN-γ responses. IL-2 and TNF-α in serum from immunized mice were extremely significantly higher than control mice (P<0.01) and IFN-γ was significantly higher than that of control mice (P<0.05).Additionally, the proliferation of lymphocytes from spleen of mice immunized with recombinant E protein was stimulated. The results indicated that recombinant GTV E protein could effectively stimulate humoral and lymphocyte immune responses in mice. Moreover, when mice were challenged with GTV, by using Real-time RT-PCR, the loads of virus in tissues from mice immunized with recombinant E protein were significantly decreased, revealing that recombinant E protein contributed to protection against GTV challenge. This study demonstrated that the E. coli-expressed recombinant GTV E protein could serve as a potential subunit vaccine candidate against GTV and provided a theoretical basis for the prevention and control of Tembusu virus.

Key words: Tembusu virus, Envelope protein, Immune response, Subunit vaccine

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• S432.4+2