Expression and immunogenicity of human immunodeficiency virus type 1 Gag expressed by a replication-competent rhabdovirus-based vaccine vector
Expression and immunogenicity of human immunodeficiency virus type 1 Gag expressed by a replication-competent rhabdovirus-based vaccine vector
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DOI：
10.1128/jvi.75.18.8724-8732.2001
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发表时间：
2001-09-01
影响因子：
5.4
通讯作者：
Schnell, MJ
中科院分区：
文献类型：
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作者：
McGettigan, JP;Sarma, S;Schnell, MJ
以人类免疫缺陷病毒1型(HIV-1)Gag蛋白为表达载体，构建了具有复制能力的横纹病毒载体，并对其在小鼠体内诱导细胞免疫应答进行了研究。我们之前描述了一种基于狂犬病病毒(RV)疫苗株的表达HIV-1gp160的载体。重组轮状病毒能够在小鼠中诱导针对HIV-1包膜蛋白的强大体液和细胞免疫反应(M.J.Schnell等人，Proc.娜塔莉。阿卡德。SCI。美国97：3544-3549,2000；J.P.McGettigan等，J.Virol。75：4430-4434,2001)。最近的研究表明，HIV-1 Gag蛋白是细胞介导的宿主免疫防御的另一个重要靶点。在这里，我们发现HIV-1Gag可以通过轮状病毒在人和非人类细胞系上有效地表达。用表达HIV-1 Gag的重组轮状病毒感染HeLa细胞后，免疫染色和Western blotting均显示HIV-1前体蛋白P55得到了高效表达。此外，HIV-1p24抗原捕获酶联免疫吸附试验和电子显微镜显示，感染细胞上清液中除弹状RV颗粒外，还有效地释放了HIV-1病毒样颗粒。为了初步筛选这种新疫苗载体的免疫原性，BALB/c小鼠接受了表达HIV-1 Gag的重组RV的单次免疫。免疫小鼠产生了针对HIV-1 Gag的CDS细胞毒性T淋巴细胞反应。此外，表达HIV-1 Gag的RV免疫小鼠后，26.8%的CD8(+)T细胞在与表达HIV-1 Gag的重组痘苗病毒攻击后产生γ-干扰素。这些结果进一步证实和扩展了基于RV的载体作为潜在的HIV-1疫苗的效力。
A replication-competent rhabdovirus-based vector expressing human immunodeficiency virus type 1 (HIV-1) Gag protein was characterized on human cell lines and analyzed for the induction of a cellular immune response in mice. We previously described a rabies virus (RV) vaccine strain-based vector expressing HIV-1 gp160. The recombinant RV was able to induce strong humoral and cellular immune responses against the HIV-1 envelope protein in mice (M. J. Schnell et al., Proc. Natl. Acad. Sci. USA 97:3544-3549, 2000; J. P. McGettigan et al., J. Virol. 75:4430-4434, 2001). Recent research suggests that the HIV-1 Gag protein is another important target for cell-mediated host immune defense. Here we show that HIV-1 Gag can efficiently be expressed by RV on both human and nonhuman cell lines. Infection of HeLa cells with recombinant RV expressing HIV-1 Gag resulted in efficient expression of HIV-1 precursor protein p55 as indicated by both immunostaining and Western blotting. Moreover, HIV-1 p24 antigen capture enzyme-linked immunosorbent assay and electron microscopy showed efficient release of HIV-1 virus-like particles in addition to bullet-shaped RV particles in the supernatants of the infected cells. To initially screen the immunogenicity of this new vaccine vector, BALB/c mice received a single vaccination with the recombinant RV expressing HIV-1 Gag. Immunized mice developed a vigorous CDS' cytotoxic T-lymphocyte response against HIV-1 Gag. In addition, 26.8% of CD8(+) T cells from mice immunized with RV expressing HIV-1 Gag produced gamma interferon after challenge with a recombinant vaccinia virus expressing HIV-1 Gag. These results further confirm and extend the potency of RV-based vectors as a potential HIV-1 vaccine.