多价多表位流感核酸疫苗的构建及实验免疫研究

发布时间：2018-04-25 12:22

本文选题：流感病毒 + 表位　；参考：《吉林大学》2008年硕士论文

【摘要】： 本研究根据当前流感的流行现状和多个亚型高致病性禽流感病毒跨种传播的现实,成功应用生物信息学理论筛选出流感病毒的多个优势抗原表位(流感病毒内部保守的M、NP蛋白CTL表位,和H1、H7、H9 HA的Th细胞表位),设计并合成了针对多个亚型流感病毒的通用多表位基因盒Epi I和Epi II。以pVAX1真核表达质粒为载体构建与筛选出共表达NP与Epi I基因盒的DNA重组体pVAX1-NP-Epi I,共表达H5、H3亚型IV HA与Epi II基因的DNA重组体pVAX1-H5HA-Epi II-H3HA1-M2e。应用BALB/c小鼠动物模型进行了免疫原性初步研究,免疫结果显示重组DNA疫苗诱导机体产生了抗流感H1、H3、H5、H7和H9 HA的特异性体液和细胞免疫,其中诱导的H5HA、H3HA1抗体水平较高;淋巴细胞转化试验、淋巴细胞亚类FACS检测和γIFN-ELISPOT细胞免疫指标检测显示,多表位重组体均诱导了较强的细胞免疫,说明所构建的DNA重组体有良好的免疫原性,为最终获得能同时预防多种亚型流感病毒的多价疫苗奠定了基础。
[Abstract]:According to the current epidemic situation of influenza and the interspecies transmission of several subtypes of highly pathogenic avian influenza virus, Several dominant antigenic epitopes of influenza virus (CTL epitopes of MNP protein in influenza virus) were successfully screened by bioinformatics theory. The universal multiepitope gene cassette Epi I and Epi II for multiple subtypes of influenza virus were designed and synthesized. Using pVAX1 eukaryotic expression plasmid as the vector, the DNA recombinant pVAX1-NP-Epi I was constructed and screened, and the DNA recombinant pVAX1-H5HA-Epi II-H3HA1-M2eof H5H3 subtype IV HA and Epi II gene was coexpressed. The immunogenicity of BALB/c mouse model was studied. The results showed that the recombinant DNA vaccine induced specific humoral and cellular immunity against H1H3H3H5H5H7 and H9HA, and the antibody level of H5H3HA1 was higher than that of H9HA. Lymphocyte transformation test, lymphocyte subclass FACS detection and 纬 IFN-ELISPOT cell immune index detection showed that the multiepitope recombinant induced strong cellular immunity, indicating that the constructed DNA recombinant had good immunogenicity. It lays the foundation for obtaining multivalent vaccine which can prevent multiple subtypes of influenza virus at the same time.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R392

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