Isaguliants Maria G, Petrakova Natalia V, Kashuba Elena V, Suzdaltzeva Yulia G, Belikov Sergey V, Mokhonov Vladislav V, Prilipov Alexei G, Matskova Ludmila, Smirnova Irina S, Jolivet-Reynaud Colette, Nordenfelt Erik
Ivanovsky Institute of Virology, 123 098 Moscow, Russia.
Vaccine. 2004 Apr 16;22(13-14):1656-65. doi: 10.1016/j.vaccine.2003.09.047.
Numerous attempts to induce immunity against HCV core (HCV-C) by DNA immunization met serious difficulties in optimizing T-helper cell and antibody responses. Immunomodulatory properties of HCV-C could be blamed that seem to be dependent on the genotype of HCV source. Here, we characterized HCV-C gene from HCV 1b isolate 274933RU. Eukaryotic expression of HCV-C was effectively driven by CMVIE, while human elongation factor 1 alpha promoter directed low levels of HCV-C expression. C57BL/6 mice were immunized with CMVIE-driven HCV-C gene, and assessed for specific antibody production, T-cell proliferation and cytokine secretion. The number and proportion of CD19+, CD3+, CD3+/CD4+, and CD3+/CD8+ splenocytes in HCV-C gene recipients was evaluated by flow cytometry. A significant mounting drop in CD3+/CD4+ T-cell counts occurred in HCV-C gene-recipients as compared to the controls. Despite that, 75% of mice exhibited core-specific cellular reactivity revealed as high proliferative responses to HCV-C and HCV-C peptides. Stimulated T-cells secreted predominantly IFN-gamma and IL-2. A shift of epitope specificity was observed with the early response being broad, and the late limited to the HCV-C C-terminus. Thus, we demonstrate both T-cell immunogenicity and T-cell modulation by core of HCV 1b. Immune modulation by HCV core may affect host ability to mount long-lasting cellular and antibody response and should be dealt with in designing core-based HCV vaccines.
通过DNA免疫诱导针对丙型肝炎病毒核心蛋白（HCV-C）的免疫反应的众多尝试，在优化T辅助细胞和抗体反应方面遇到了严重困难。HCV-C的免疫调节特性可能是原因所在，这似乎取决于HCV来源的基因型。在此，我们对来自HCV 1b分离株274933RU的HCV-C基因进行了表征。CMVIE可有效驱动HCV-C的真核表达，而人延伸因子1α启动子指导的HCV-C表达水平较低。用CMVIE驱动的HCV-C基因免疫C57BL/6小鼠，并评估其特异性抗体产生、T细胞增殖和细胞因子分泌情况。通过流式细胞术评估HCV-C基因受体中CD19 +、CD3 +、CD3 + / CD4 +和CD3 + / CD8 +脾细胞的数量和比例。与对照组相比，HCV-C基因受体中的CD3 + / CD4 + T细胞计数显著下降。尽管如此，75%的小鼠表现出核心特异性细胞反应性，表现为对HCV-C和HCV-C肽的高增殖反应。受刺激的T细胞主要分泌IFN-γ和IL-2。观察到表位特异性发生了转变，早期反应广泛，后期则局限于HCV-C的C末端。因此，我们证明了HCV 1b核心蛋白具有T细胞免疫原性和T细胞调节作用。HCV核心蛋白的免疫调节可能会影响宿主产生持久细胞和抗体反应的能力，在设计基于核心蛋白的HCV疫苗时应予以考虑。
