Department of Molecular and Cellular Biology, Centro Nacional de Biotecnología, Consejo Superior de Investigaciones Científicas (CNB-CSIC), Campus de Cantoblanco, 28049 Madrid, Spain.
Biocomputing Unit, Centro Nacional de Biotecnología, Consejo Superior de Investigaciones Científicas (CNB-CSIC), Campus de Cantoblanco, 28049 Madrid, Spain.
Viruses. 2019 Feb 16;11(2):160. doi: 10.3390/v11020160.
The development of an effective Human Immunodeficiency Virus (HIV) vaccine that is able to stimulate both the humoral and cellular HIV-1-specific immune responses remains a major priority challenge. In this study, we described the generation and preclinical evaluation of single and double modified vaccinia virus Ankara (MVA)-based candidates expressing the HIV-1 clade C membrane-bound gp145(ZM96) trimeric protein and/or the Gag(ZM96)-Pol-Nef(CN54) (GPN) polyprotein that was processed to form Gag-induced virus-like particles (VLPs). In vitro characterization of MVA recombinants revealed the stable integration of HIV-1 genes without affecting its replication capacity. In cells that were infected with Env-expressing viruses, the gp145 protein was inserted into the plasma membrane exposing critical epitopes that were recognized by broadly neutralizing antibodies (bNAbs), whereas Gag-induced VLPs were released from cells that were infected with GPN-expressing viruses. VLP particles as well as purified MVA virions contain Env and Gag visualized by immunoelectron microscopy and western-blot of fractions that were obtained after detergent treatments of purified virus particles. In BALB/c mice, homologous MVA-gp145-GPN prime/boost regimen induced broad and polyfunctional Env- and Gag-specific CD4 T cells and antigen-specific T follicular helper (Tfh) and Germinal Center (GC) B cells, which correlated with robust HIV-1-specific humoral responses. Overall, these results support the consideration of MVA-gp145-GPN vector as a potential vaccine candidate against HIV-1.
开发一种能够刺激体液和细胞 HIV-1 特异性免疫反应的有效人类免疫缺陷病毒 (HIV) 疫苗仍然是一个主要的优先挑战。在这项研究中,我们描述了表达 HIV-1 群 C 膜结合 gp145(ZM96)三聚体蛋白和/或 Gag(ZM96)-Pol-Nef(CN54)(GPN)多蛋白的单一和双重改良安卡拉痘苗病毒 (MVA) 候选物的产生和临床前评估,该多蛋白被加工形成 Gag 诱导的病毒样颗粒 (VLPs)。MVA 重组体的体外特性分析表明,HIV-1 基因的稳定整合不会影响其复制能力。在表达Env 的病毒感染的细胞中,gp145 蛋白插入质膜,暴露出被广泛中和抗体 (bNAb) 识别的关键表位,而 Gag 诱导的 VLPs 从感染 GPN 表达病毒的细胞中释放出来。电镜免疫和经去污剂处理后获得的纯化病毒粒子的 Western blot 分析表明,VLP 颗粒以及纯化的 MVA 病毒粒子都含有 Env 和 Gag。在 BALB/c 小鼠中,同源 MVA-gp145-GPN 初免/加强方案诱导了广泛的和多功能的 Env 和 Gag 特异性 CD4 T 细胞以及抗原特异性滤泡辅助 T 细胞 (Tfh) 和生发中心 (GC) B 细胞,这与强大的 HIV-1 特异性体液反应相关。总的来说,这些结果支持将 MVA-gp145-GPN 载体作为一种针对 HIV-1 的潜在候选疫苗。
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