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构建西尼罗病毒（West nile virus,WNV）与基孔肯雅病毒（Chikungunya virus, CHIKV）的新型双价重组DNA疫苗并在小鼠中评价其免疫原性。本研究以WNV的prM-E蛋白和CHIKV的C-E3-E2-6K-E1或E3-E2-6K-E1蛋白为靶抗原，构建了WNV和CHIKV双价重组DNA疫苗VRC-PrME-2A-CE和VRC-PrME-2A-E。转染HEK293T细胞后通过Western blot验证目的蛋白的表达。两剂次同源免疫小鼠，通过ELISA和Elispot进行体液与细胞免疫检测，并与表达WNV-XJ11129-3株的prM-E蛋白的WNV单价DNA疫苗VRC-prME和表达CHIKV 181/clone25的C-E3-E2-6K-E1蛋白的CHIKV单价DNA疫苗VRC-CE进行比较。结果显示，双价DNA疫苗构建成功。重组双价DNA疫苗单剂或加强免疫小鼠后均能诱导较高水平抗原特异性IgG，且与单价疫苗滴度相当。细胞免疫检测结果显示：加强免疫后VRC-PrME-2A-CE表现出针对CHIKV E1和CHIKV E2最高的分泌IFN-γ的T细胞水平，但与单价疫苗VRC-CE无统计学差异。本研究成功构建两款针对WNV和CHIKV的双价DNA疫苗，免疫小鼠后可以诱导产生良好的体液和细胞免疫应答，本研究为新型WNV和CHIKV双价基因工程疫苗的研发应用提供了实验参考。
Abstract：To develop novel bivalent recombinant DNA vaccines against both West Nile Virus(WNV) and Chikungunya virus(CHIKV),then evaluate its immunogenicity in mice. In this study, the prME protein of WNV and the C-E3-E2-6K-E1 or E3-E2-6K-E1 protein were used as target antigens to construct the bivalent recombinant DNA vaccines, named VRC-prME-2A-CE and VRC-prME-2A-E. The expression of the target proteins was confirmed by Western blot after transfection of HEK 293T cells. Mice were immunized twice via intramuscularly with these DNA vaccines, and humoral and cellular immune responses were assessed by ELISA and Elispot, respectively. The results were compared with those of the monovalent DNA vaccines VRC-prME(expressing the prME protein alone) and VRC-CE(expressing the C-E3-E2-6K-E1 protein alone). The results showed that the bivalent DNA vaccines were successfully constructed. Both single and booster immunizations with the bivalent DNA vaccines induced high levels of antigen-specific IgG in mice, comparable to those induced by the monovalent vaccines. Elispot assays revealed that the bivalent vaccines VRC-prME-2A-CE exhibited the highest cellular immune response levels against CHIKV E1 and CHIKV E2 after booster immunization, but there is no statistical difference compared to the monovalent vaccine VRC-CE.. In conclusion, two novel bivalent DNA vaccines targeting WNV and CHIKV were successfully developed. These vaccines induced robust humoral and cellular immune responses in mice, providing experimental evidence for the development and application of novel vaccines against WNV and CHIKV.
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基本信息:
DOI：10.13242/j.cnki.bingduxuebao.250137
中图分类号:R392
引用信息:
[1]舒畅,李梦哲,翟程程,等.西尼罗病毒和基孔肯雅病毒双价重组DNA疫苗的构建及在小鼠中诱导的免疫应答研究[J].病毒学报,2025,41(03):713-724.DOI:10.13242/j.cnki.bingduxuebao.250137.
基金信息:
国家重点研发计划项目（2022YFC2304100）,题目：新冠与流感等大流行风险病原体多肽疫苗的研制~~
2025-04-16
2025
2025-04-30
2025-06-03
2025
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2025-05-14
2025-05-14
2025-05-14