Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, Colorado, USA.
J Virol. 2013 Jul;87(14):7940-51. doi: 10.1128/JVI.00752-13. Epub 2013 May 8.
Feline immunodeficiency virus (FIV) is a lentivirus that causes AIDS in domestic cats, similar to human immunodeficiency virus (HIV)/AIDS in humans. The FIV accessory protein Vif abrogates the inhibition of infection by cat APOBEC3 restriction factors. FIV also encodes a multifunctional OrfA accessory protein that has characteristics similar to HIV Tat, Vpu, Vpr, and Nef. To examine the role of vif and orfA accessory genes in FIV replication and pathogenicity, we generated chimeras between two FIV molecular clones with divergent disease potentials: a highly pathogenic isolate that replicates rapidly in vitro and is associated with significant immunopathology in vivo, FIV-C36 (referred to here as high-virulence FIV [HV-FIV]), and a less-pathogenic strain, FIV-PPR (referred to here as low-virulence FIV [LV-FIV]). Using PCR-driven overlap extension, we produced viruses in which vif, orfA, or both genes from virulent HV-FIV replaced equivalent genes in LV-FIV. The generation of these chimeras is more straightforward in FIV than in primate lentiviruses, since FIV accessory gene open reading frames have very little overlap with other genes. All three chimeric viruses exhibited increased replication kinetics in vitro compared to the replication kinetics of LV-FIV. Chimeras containing HV-Vif or Vif/OrfA had replication rates equivalent to those of the virulent HV-FIV parental virus. Furthermore, small interfering RNA knockdown of feline APOBEC3 genes resulted in equalization of replication rates between LV-FIV and LV-FIV encoding HV-FIV Vif. These findings demonstrate that Vif-APOBEC interactions play a key role in controlling the replication and pathogenicity of this immunodeficiency-inducing virus in its native host species and that accessory genes act as mediators of lentiviral strain-specific virulence.
猫免疫缺陷病毒(FIV)是一种慢病毒,可导致家猫患艾滋病,类似于人类免疫缺陷病毒(HIV)/人类艾滋病。FIV 的辅助蛋白 Vif 消除了猫 APOBEC3 限制因子对感染的抑制作用。FIV 还编码一种多功能 OrfA 辅助蛋白,具有类似于 HIV Tat、Vpu、Vpr 和 Nef 的特征。为了研究 vif 和 orfA 辅助基因在 FIV 复制和致病性中的作用,我们在两个具有不同疾病潜力的 FIV 分子克隆之间生成嵌合体:一种高度致病的分离株,在体外快速复制,并与体内显著的免疫病理学相关,FIV-C36(这里称为高毒力 FIV [HV-FIV])和一种低致病性株,FIV-PPR(这里称为低毒力 FIV [LV-FIV])。使用 PCR 驱动的重叠延伸,我们生成了病毒,其中来自 HV-FIV 的 vif 或 orfA 或这两个基因替换了 LV-FIV 中的相应基因。与灵长类慢病毒相比,FIV 更容易产生这些嵌合体,因为 FIV 辅助基因开放阅读框与其他基因的重叠很少。与 LV-FIV 的复制动力学相比,所有三种嵌合病毒在体外的复制动力学都有所增加。含有 HV-Vif 或 Vif/OrfA 的嵌合体的复制率与毒力 HV-FIV 亲本病毒相当。此外,猫 APOBEC3 基因的小干扰 RNA 敲低导致 LV-FIV 和编码 HV-FIV Vif 的 LV-FIV 之间的复制率均等化。这些发现表明,Vif-APOBEC 相互作用在控制这种免疫缺陷诱导病毒在其天然宿主物种中的复制和致病性方面发挥着关键作用,并且辅助基因是调节慢病毒株特异性毒力的介质。