Comparison of the protective efficacy of DNA and baculovirus-derived protein vaccines for EBOLA virus in guinea pigs
Comparison of the protective efficacy of DNA and baculovirus-derived protein vaccines for EBOLA virus in guinea pigs
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DOI：
10.1016/s0168-1702(02)00338-6
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发表时间：
2003-04-01
期刊：
影响因子：
5
通讯作者：
Schmaljohn, CS
中科院分区：
文献类型：
--
作者：
Mellquist-Riemenschneider, JL;Garrison, AR;Schmaljohn, CS
丝状病毒埃博拉病毒（EBOV）和马尔堡病毒（MARV）引起人类严重出血热，目前尚无疫苗。在此之前，研究发现，先用表达MARV糖蛋白（GP）基因的DNA疫苗启动剂量，然后用杆状病毒衍生的重组GP蛋白增强，可使豚鼠获得保护性免疫（Hevey等人，2001年）。疫苗20,568-593)。为了确定类似的初始-增强疫苗方法是否对EBOV有效，我们产生并鉴定了表达全长EBOV GP（GP(1,2)）或末端缺失GP（GPa-）的重组杆状病毒，并检测了它们在豚鼠中的免疫原性。正如预期的那样，感染GPa-重组蛋白的细胞比感染GP（1,2）重组蛋白的细胞分泌更多的GP(1)。在凝集素结合研究中，发现昆虫细胞培养衍生的病毒粒子GP与哺乳动物细胞衍生的病毒粒子GP不同，因为它们没有复杂/杂交的n链聚糖或含有唾液酸的聚糖。尽管存在这些差异，杆状病毒衍生的GP能够将单克隆抗体结合到EBOV GP上的五个不同的表位上，这表明蛋白质的抗原结构保持完整。作为杆状病毒衍生蛋白引发细胞介导免疫反应能力的衡量标准，我们在培养的人树突状细胞中评估了GPa-蛋白对t细胞的刺激能力。与对照组相比，细胞毒性的增加表明杆状病毒蛋白有能力引起细胞介导的免疫反应。单独接种杆状病毒衍生的GPs或DNA引物-杆状病毒蛋白增强方案的豚鼠通过ELISA和斑块减少中和试验检测产生抗体反应；然而，当蛋白质单独或与DNA疫苗结合使用时，保护作用不完全。这些数据表明，对MARV有效的疫苗方法对豚鼠的EBOV无效。Elsevier Science B.V.出版
The filoviruses Ebola virus (EBOV) and Marburg virus (MARV) cause severe hemorrhagic fever in humans for which no vaccines are available. Previously, a priming dose of a DNA vaccine expressing the glycoprotein (GP) gene of MARV followed by boosting with recombinant baculovirus-derived GP protein was found to confer protective immunity to guinea pigs (Hevey et al., 2001. Vaccine 20, 568-593). To determine whether a similar prime-boost vaccine approach would be effective for EBOV, we generated and characterized recombinant baculoviruses expressing full-length EBOV GP (GP(1,2)) or a terminally-deleted GP (GPa-) and examined their immunogenicity in guinea pigs. As expected, cells infected with the GPa- recombinant secreted more GP(1) than those infected with the GP(1,2) recombinant. In lectin binding studies, the insect cell culture-derived GPs were found to differ from mammalian cell derived virion GP, in that they had no complex/hybrid N-linked glycans or glycans containing sialic acid. Despite these differences, the baculovirus-derived GPs were able to bind monoclonal antibodies to five distinct epitopes on EBOV GP, indicating that the antigenic structures of the proteins remain intact. As a measure of the ability of the baculovirus-derived proteins to elicit cell-mediated immune responses, we evaluated the T-cell stimulatory capacity of the GPa- protein in cultured human dendritic cells. Increases in cytotoxicity as compared to controls suggest that the baculovirus proteins have the capacity to evoke cell-mediated immune responses. Guinea pigs vaccinated with the baculovirus-derived GPs alone, or in a DNA prime-baculovirus protein boost regimen developed antibody responses as measured by ELISA and plaque reduction neutralization assays; however, incomplete protection was achieved when the proteins were given alone or in combination with DNA vaccines. These data indicate that a vaccine approach that was effective for MARV is not effective for EBOV in guinea pigs. Published by Elsevier Science B.V.