天坛痘病毒载体改造及其对HIV重组疫苗免疫原性的影响

发布时间：2018-12-14 03:40

【摘要】： 痘病毒因其能诱导持久的体液免疫应答和强大的细胞免疫应答、在细胞质中复制而无整合入基因组的危险、可容纳大片段的外源基因等优点,被应用于HIV疫苗研究中。而提高痘病毒载体疫苗激起的免疫应答和克服预存载体反应一直是痘病毒载体疫苗临床试验中有待解决的问题。本研究选择HIV-1 CN54的gag-pol和env作为免疫原,引入小鼠的IL-12、IL-15、GM-CSF后检测痘病毒重组疫苗的免疫原性。结果是三种细胞因子没有影响细胞和体液免疫应答的强度。细胞因子mIL-15延长了痘病毒载体疫苗激起的细胞反应时间；mGM-CSF延长了痘病毒载体疫苗激起的体液反应时间并小幅度延长了细胞免疫应答持续时间；细胞因子mIL-12、VTKgpemIL-15+ VTKgpemGM-CSF联合免疫和VTKgpemIL-12和VTKgpemGM-CSF联合免疫都对细胞和体液免疫无影响。缺失A33能抑制胞间传播,增加EEV的分泌,增强胞外远距离传播。改变体内传播方式,也许能克服预存载体反应。实验证明,缺失A33能部分克服预存载体反应。同时,在未实验小鼠中VTKgpe△A33诱导出更高的体液免疫应答。并在VTKgpe二次加强后,由于前次免疫未产生A33抗体而增强VTKgpe胞间传播,一定程度加强了细胞免疫应答和抑制了针对载体的体液免疫应答。本研究为最终获得免疫原性好和能克服抗载体反应的复制型天坛载体打下了基础。
[Abstract]:Because of its ability to induce persistent humoral immune response and strong cellular immune response, to replicate in the cytoplasm without the risk of integration into the genome, and to accommodate large fragments of foreign genes, poxvirus has been used in the study of HIV vaccine. Improving the immune response and overcoming the preexisting vector response are the problems to be solved in the clinical trials of poxvirus vector vaccine. In this study, gag-pol and env of HIV-1 CN54 were selected as immunogen, and the immunogenicity of recombinant vaccine of poxvirus was detected by introducing IL-12,IL-15,GM-CSF of mice. The result was that the three cytokines did not affect the intensity of cellular and humoral immune responses. The cytokine mIL-15 prolonged the cell reaction time stimulated by the vector vaccine, mGM-CSF prolonged the humoral reaction time and prolonged the cell immune response time to a small extent. Cytokine mIL-12,VTKgpemIL-15 VTKgpemGM-CSF combined immunization and VTKgpemIL-12 and VTKgpemGM-CSF combined immunization had no effect on cellular and humoral immunity. Deletion of A 33 inhibited intercellular transmission, increased EEV secretion and enhanced extracellular long distance transmission. Changing the mode of transmission in vivo may overcome the pre-storage carrier reaction. Experiments show that missing A 33 can partially overcome the pre-storage carrier reaction. At the same time, VTKgpe A 33 induced a higher humoral immune response in untested mice. After the second strengthening of VTKgpe, the cellular immune response of VTKgpe was enhanced and the humoral immune response against the vector was inhibited to a certain extent because the previous immunization did not produce A33 antibody. This study laid a foundation for obtaining the replicative Temple of Heaven vector which has good immunogenicity and can overcome the anti-carrier reaction.
【学位授予单位】：中国疾病预防控制中心
【学位级别】：博士
【学位授予年份】：2010
【分类号】：R392

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