Molecular and Immunological Studies on Feline Immunodeficiency Virus

猫免疫缺陷病毒的分子和免疫学研究

基本信息

批准号：
07306014
负责人：
MIKAMI Takeshi
金额：
$ 8.9万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07306014/
关键词：
FIV evolution quantification neutralizing antibody AP-1 vif ORF-A CD8 ORF-1

项目摘要

The purpose of the project is to clarify the genetic properties of feline immunodeficiency virus (FIV) and to know the pathogenetic mechanism of the virus in vivo by studying comprehensively from the molecular biological and immunological aspects. The study is still continuing but we obtained the following results.1) We isolated four and five FIV strains from cats in Taiwan and Argentina, respectively. Sequence analyzes of their env V3-V5 region revealed that Taiwanese isolates belonged to subtype C and four of the five Argentine isolates formad a new cluster designated as subtype E.2) We established novel methods to measure both the viral burden in the peripheral blood mononuclear cells and the neutralizing antibody titer against FIV.And then, we applied the methods to analyzes of FIV-infected cats.3) We obtained monoclonal antibodies (mAbs) recognizing feline CD4 and CD8 alpha molecules. Further, we cloned the cDNA of feline CD8 alpha and beta chains and expressed them in COS-7 cells … More , and analyzed the sequences of the cDNA and the mAbs.4) We cloned genes of feline Fas antigen and Fas ligand. The Fas gene was 942 bp long and has a homology with human and murine homologues at 55.6 and 47.9% at amino acid level, respectively. The Fas ligand gene was 840 bp long and has a homology with human and murine homologues at 88.7 and 78.2% at amino acid level, respectively. The Fas antigen and the ligand were expressed in various tissues and several cell lines. Further we obtained some evidence that the Fas antigen-ligand system was associated with the apoptosis observed in FIV-infected cats.5) We constructed FIV mutants which deleted AP-1 binding site, vif and ORF-A genes, and inoculated them into SPF cats. We found that vif gene is quite important for the viral infectivity and growth ability in vivo. On the other hand, we also found that the AP-1 binding site has a little responsibility for the viral growth ability, and ORF-A gene is dispensable but responsible for the viral growth ability significantly in vivo. Less

该项目的目的是阐明猫免疫缺陷病毒（FIV）的遗传特性，并通过从分子生物学和免疫学方面进行彻底研究，了解体内病毒的致病机制。这项研究仍在继续，但我们获得了以下结果。1）我们分别从台湾和阿根廷的猫分离了四个和五个FIV菌株。对其Env V3-V5区域的序列分析表明，台湾分离株属于亚型C，而五个阿根廷分离株中的四个则形成了一个新的簇，该簇被指定为亚型E.2），我们建立了新的方法，以测量外周核细胞中的病毒性燃烧，并在外周核细胞和中性抗体中对抗体的中和抗体进行了反对的方法，我们对fiv的中性序列进行了分析。我们获得了识别猫CD4和CD8α分子的单克隆抗体（mAb）。此外，我们克隆了猫CD8α和β链的cDNA，并在COS-7细胞中表达了它们……更多，并分析了cDNA和mabs的序列。4）我们克隆了猫Feline Fash抗原和Fas配体的基因。 FAS基因的长度为942 bp，在氨基酸水平上的人和鼠同源物的同源物分别为55.6和47.9％。 FAS配体基因长840 bp，并且在氨基酸水平上的人和鼠同源物的同源物分别为88.7和78.2％。 FAS抗原和配体以各种时间和几个细胞系表示。此外，我们获得了一些证据，表明FAS抗原配体系统与FIV感染的CAT中观察到的凋亡有关。5）我们构建了FIV突变体，这些突变体删除了AP-1结合位点，VIF和ORF-A基因，并将其接种到SPF CAT中。我们发现VIF基因对于体内病毒感染和生长能力非常重要。另一方面，我们还发现，AP-1结合位点对病毒生长能力有很少的责任，ORF-A基因具有可分配的，但负责体内病毒生长能力。较少的