Characterization of a Single-Cycle Rabies Virus-Based Vaccine Vector
Characterization of a Single-Cycle Rabies Virus-Based Vaccine Vector
复制标题
DOI：
10.1128/jvi.01870-09
复制
发表时间：
2010-03-01
影响因子：
5.4
通讯作者：
Schnell, Matthias J.
中科院分区：
文献类型：
--
作者：
Gomme, Emily A.;Faul, Elizabeth J.;Schnell, Matthias J.
基于狂犬病病毒(RV)的重组载体已经在小鼠和猴子模型中证明了它们在产生长期的抗原特异性免疫反应方面的有效性。然而，由于载体的致病性，具有复制能力的病毒载体构成了重大的安全问题。轮状病毒的致病性在很大程度上归因于其糖蛋白(RV-G)，它促进了RV的附着和进入宿主细胞。我们通过从表达HIV-1 Gag的RV疫苗载体(SPBN-Delta G-Gag)中缺失G基因，研制出了一种活的单周期RV。在稳定表达RV-G的细胞上通过SPBN-Delta G-GAG可以有效地增殖G缺失的RV。比较单循环RV和复制能力对照(BNSP-GAG)的体内免疫原性数据显示，RV特异性抗体较低；然而，两种疫苗载体的总体同型特征(IgG2a/IgG1)相似。尽管有这种差异，免疫SPBN-Delta G-Gag和BNSP-Gag的小鼠获得了类似水平的Gag特异性CD8(+)T细胞反应，通过主要组织相容性复合体I类Gag-四聚体染色、伽马干扰素-酶联免疫斑点试验和细胞毒性T细胞试验来测量。此外，对于两种轮状病毒疫苗载体，这些细胞反应在低至103个病灶形成单位的免疫滴度下保持相同。CD8(+)T细胞反应通过单周期RV和异种水泡性口炎病毒糖蛋白的增强而显著增强。这些发现表明，单周期轮状病毒是未来开发HIV-1和其他传染病疫苗的一种有效的替代复制能力的轮状病毒载体。
Recombinant rabies virus (RV)-based vectors have demonstrated their efficacy in generating long-term, antigen-specific immune responses in murine and monkey models. However, replication-competent viral vectors pose significant safety concerns due to vector pathogenicity. RV pathogenicity is largely attributed to its glycoprotein (RV-G), which facilitates the attachment and entry of RV into host cells. We have developed a live, single-cycle RV by deletion of the G gene from an RV vaccine vector expressing HIV-1 Gag (SPBN-Delta G-Gag). Passage of SPBN-Delta G-Gag on cells stably expressing RV-G allowed efficient propagation of the G-deleted RV. The in vivo immunogenicity data comparing single-cycle RV to a replication-competent control (BNSP-Gag) showed lower RV-specific antibodies; however, the overall isotype profiles (IgG2a/IgG1) were similar for the two vaccine vectors. Despite this difference, mice immunized with SPBN-Delta G-Gag and BNSP-Gag mounted similar levels of Gag-specific CD8(+) T-cell responses as measured by major histocompatibility complex class I Gag-tetramer staining, gamma interferon-enzyme-linked immunospot assay, and cytotoxic T-cell assay. Moreover, these cellular responses were maintained equally at immunization titers as low as 103 focus-forming units for both RV vaccine vectors. CD8(+) T-cell responses were significantly enhanced by a boost with a single-cycle RV complemented with a heterologous vesicular stomatitis virus glycoprotein. These findings demonstrate that single-cycle RV is an effective alternative to replication-competent RV vectors for future development of vaccines for HIV-1 and other infectious diseases.