Moldoveanu Z, Porter D C, Lu A, McPherson S, Morrow C D
Department of Microbiology, University of Alabama at Birmingham 35294, USA.
Vaccine. 1995 Aug;13(11):1013-22. doi: 10.1016/0264-410x(95)00018-v.
Several viruses have been exploited for the development of recombinant vaccine vectors in which to express foreign proteins. Recently, we have described a system utilizing the RNA virus, poliovirus. We have constructed poliovirus genomes in which regions of the capsid have been substituted with gene fragments of the HIV gag and env genes. A complementation system has been designed to encapsidate defective genomes by providing the capsid protein in trans from a recombinant vaccinia virus (VV-P1). Serial passage in the presence of VV-P1 resulted in the generation of stocks of these encapsidated replicons. Infection of cells with these encapsidated replicons resulted in the expression of the recombinant protein as a fusion protein with the poliovirus capsid proteins VP4 and VP1. In this study, we have utilized encapsidated replicons which express the HIV-1-gag capsid protein (p24) as well as 1.5 kb of the HIV-1 env gene. Stocks of these encapsidated replicons were obtained by 20 serial passages in the presence of VV-P1. In addition, passage of the encapsidated replicons in the presence of poliovirus type 2 Lansing resulted in the encapsidation of the replicons by the capsid proteins provided by poliovirus. The administration of the type 2 Lansing/encapsidated replicons expressing HIV-1 gag in BALB/c mice by intramuscular, intrarectal, or intragastric routes resulted in the generation of antibodies in the serum and secretions against both poliovirus and HIV-1 gag. To prove that the replicons alone are immunogenic, we administered replicons expressing either HIV-1 gag or env to transgenic mice which expressed the receptor for poliovirus type 1. Immunization of these mice by the intramuscular route resulted in the generation of serum antibodies specific for poliovirus as well as for HIV-1 antigens. The results obtained led us to the conclusion that the replicons are immunogenic when given alone or in the presence of poliovirus. These results are important for the use of the poliovirus replicons as a recombinant vaccine vector.
几种病毒已被用于开发用于表达外源蛋白的重组疫苗载体。最近，我们描述了一种利用RNA病毒脊髓灰质炎病毒的系统。我们构建了脊髓灰质炎病毒基因组，其中衣壳区域已被HIV gag和env基因的基因片段所取代。设计了一种互补系统，通过从重组痘苗病毒（VV-P1）反式提供衣壳蛋白来包装缺陷基因组。在VV-P1存在下连续传代导致产生这些包装好的复制子的毒株。用这些包装好的复制子感染细胞导致重组蛋白作为与脊髓灰质炎病毒衣壳蛋白VP4和VP1的融合蛋白表达。在本研究中，我们利用了表达HIV-1-gag衣壳蛋白（p24）以及1.5 kb HIV-1 env基因的包装好的复制子。这些包装好的复制子的毒株是在VV-P1存在下通过20次连续传代获得的。此外，在脊髓灰质炎病毒2型兰辛株存在下传代包装好的复制子，导致复制子被脊髓灰质炎病毒提供的衣壳蛋白包装。通过肌肉内、直肠内或胃内途径将表达HIV-1 gag的2型兰辛株/包装好的复制子给予BALB/c小鼠，导致血清和分泌物中产生针对脊髓灰质炎病毒和HIV-1 gag的抗体。为了证明单独的复制子具有免疫原性，我们将表达HIV-1 gag或env的复制子给予表达1型脊髓灰质炎病毒受体的转基因小鼠。通过肌肉内途径免疫这些小鼠导致产生针对脊髓灰质炎病毒以及HIV-1抗原的血清抗体。所获得的结果使我们得出结论，复制子单独给予或在脊髓灰质炎病毒存在下给予时具有免疫原性。这些结果对于将脊髓灰质炎病毒复制子用作重组疫苗载体很重要。
