Kucerova L, Altanerova V, Altaner C, Boris-Lawrie K
Cancer Research Institute, Slovak Academy of Sciences, SK-833 91 Bratislava, Slovakia.
J Virol. 1999 Oct;73(10):8160-6. doi: 10.1128/JVI.73.10.8160-8166.1999.
Infection with a replication-competent bovine leukemia virus structural gene vector (BLV SGV) is an innovative vaccination approach to prevent disease by complex retroviruses. Previously we developed BLV SGV that constitutively expresses BLV gag, pol, and env and related cis-acting sequences but lacks tax, rex, RIII, and GIV and most of the BLV long terminal repeat sequences, including the cis-acting Tax and Rex response elements. The novel SGV virus is replication competent and replicates a selectable vector to a titer similar to that of the parental BLV in cell culture. The overall goal of this study was to test the hypothesis that infection with BLV SGV is nonpathogenic in rabbits. BLV infection of rabbits by inoculation of cell-free BLV or cell-associated BLV typically causes an immunodeficiency-like syndrome and death by 1 year postinfection. We sought to evaluate whether in vivo transfection of BLV provirus recapitulates pathogenic BLV infection and to compare BLV and BLV SGV with respect to infection, immunogenicity, and clinical outcome. Three groups of rabbits were subjected to in vivo transfection with BLV, BLV SGV, or negative control DNA. The results of our 20-month study indicate that in vivo transfection of rabbits with BLV recapitulates the fatal BLV infection produced by cell-free or cell-associated BLV. The BLV-infected rabbits exhibited sudden onset of clinical decline and immunodeficiency-like symptoms that culminated in death. BLV and BLV SGV infected peripheral blood mononuclear cells and induced similar levels of seroconversion to BLV structural proteins. However, BLV SGV exhibited a reduced proviral load and did not trigger the immunodeficiency-like syndrome. These results are consistent with the hypothesis that BLV SGV is infectious and immunogenic and lacks BLV pathogenicity in rabbits, and they support the use of this modified proviral vector delivery system for vaccines against complex retroviruses like BLV.
用具有复制能力的牛白血病病毒结构基因载体(BLV SGV)进行感染是一种通过复杂逆转录病毒预防疾病的创新疫苗接种方法。此前我们开发了BLV SGV,其组成性表达BLV gag、pol和env以及相关的顺式作用序列,但缺乏tax、rex、RIII和GIV以及大部分BLV长末端重复序列,包括顺式作用的Tax和Rex反应元件。这种新型的SGV病毒具有复制能力,并且在细胞培养中能将一种可选择载体复制到与亲本BLV相似的滴度。本研究的总体目标是检验BLV SGV感染对兔子无致病性这一假设。通过接种无细胞BLV或细胞相关BLV对兔子进行BLV感染通常会导致类似免疫缺陷的综合征,并在感染后1年内死亡。我们试图评估BLV前病毒的体内转染是否能重现致病性BLV感染,并比较BLV和BLV SGV在感染、免疫原性和临床结果方面的差异。三组兔子分别接受了BLV、BLV SGV或阴性对照DNA的体内转染。我们为期20个月的研究结果表明,用BLV对兔子进行体内转染可重现由无细胞或细胞相关BLV产生的致命BLV感染。感染BLV的兔子表现出临床状况突然恶化和类似免疫缺陷的症状,最终导致死亡。BLV和BLV SGV感染外周血单核细胞,并诱导产生相似水平的针对BLV结构蛋白的血清转化。然而,BLV SGV的前病毒载量降低,并且未引发类似免疫缺陷的综合征。这些结果与BLV SGV具有感染性和免疫原性且在兔子中缺乏BLV致病性这一假设一致,并且支持使用这种改良的前病毒载体递送系统来开发针对像BLV这样的复杂逆转录病毒的疫苗。