Ebola virus-like particles produced in insect cells exhibit dendritic cell stimulating activity and induce neutralizing antibodies.
Ebola virus-like particles produced in insect cells exhibit dendritic cell stimulating activity and induce neutralizing antibodies.
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DOI：
10.1016/j.virol.2006.03.021
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发表时间：
2006-08
期刊：
影响因子：
3.7
通讯作者：
L. Ye;Jianguo Lin;Yuliang Sun;Soumaya Bennouna;Michael K. Lo;Qingyang Wu;Z. Bu;B. Pulendran;R. Compans;Chinglai Yang
中科院分区：
文献类型：
--
作者：
L. Ye;Jianguo Lin;Yuliang Sun;Soumaya Bennouna;Michael K. Lo;Qingyang Wu;Z. Bu;B. Pulendran;R. Compans;Chinglai Yang
产生了表达埃博拉病毒 VP40 (rBV-VP40) 或 GP (rBV-GP) 蛋白的重组杆状病毒 (rBV)。如电子显微镜所示，rBV-VP40 感染 Sf9 昆虫细胞导致细胞表面丝状颗粒的组装和出芽。通过将 rBV-VP40 和 rBV-GP 共感染 Sf9 细胞产生埃博拉病毒样颗粒 (VLP)，并通过 SDS-PAGE 和蛋白质印迹分析证明埃博拉 GP 掺入 VLP 中。昆虫细胞的重组杆状病毒感染产生了高水平的 VLP，其被证明可以刺激人类树突状细胞分泌细胞因子，类似于哺乳动物细胞中产生的 VLP。通过免疫小鼠评估昆虫细胞中产生的埃博拉病毒样颗粒的免疫原性。抗体反应分析表明，大多数 GP 特异性抗体属于 IgG2a 亚型，而没有诱导显着水平的 GP 特异性 IgG1 亚型抗体，表明诱导了 Th1 偏向的免疫反应。此外，来自埃博拉VLP免疫小鼠的血清能够在单轮感染测定中阻断埃博拉GP假型HIV病毒的感染，表明诱导了针对埃博拉GP蛋白的中和抗体。这些结果表明，使用重组杆状病毒在昆虫细胞中生产埃博拉VLP代表了开发针对埃博拉病毒感染的疫苗的一种有前途的方法。
Recombinant baculoviruses (rBV) expressing Ebola virus VP40 (rBV-VP40) or GP (rBV-GP) proteins were generated. Infection of Sf9 insect cells by rBV-VP40 led to assembly and budding of filamentous particles from the cell surface as shown by electron microscopy. Ebola virus-like particles (VLPs) were produced by coinfection of Sf9 cells with rBV-VP40 and rBV-GP, and incorporation of Ebola GP into VLPs was demonstrated by SDS-PAGE and Western blot analysis. Recombinant baculovirus infection of insect cells yielded high levels of VLPs, which were shown to stimulate cytokine secretion from human dendritic cells similar to VLPs produced in mammalian cells. The immunogenicity of Ebola VLPs produced in insect cells was evaluated by immunization of mice. Analysis of antibody responses showed that most of the GP-specific antibodies were of the IgG2a subtype, while no significant level of IgG1 subtype antibodies specific for GP was induced, indicating the induction of a Th1-biased immune response. Furthermore, sera from Ebola VLP immunized mice were able to block infection by Ebola GP pseudotyped HIV virus in a single round infection assay, indicating that a neutralizing antibody against the Ebola GP protein was induced. These results show that production of Ebola VLPs in insect cells using recombinant baculoviruses represents a promising approach for vaccine development against Ebola virus infection.