HIV-2 ENVELOPE GLYCOPROTEINS AND CELL FUSION

HIV-2 包膜糖蛋白和细胞融合

• 批准号: 3791255
• 负责人: RICHARD W COMPANS
• 金额: --
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3791255
• 关键词: AIDS; T lymphocyte; acidity /alkalinity; antibody; cell fusion; electron microscopy; gene expression; genetic manipulation; glycoproteins; human immunodeficiency virus 2; immunofluorescence technique; laboratory rabbit; membrane proteins; molecular cloning; point mutation; protein sequence; surface antigens; synthetic peptide; vaccinia virus; virus assembly; virus cytopathogenic effect; virus envelope; virus protein

Human immunodeficiency virus-2 (HIV-2) has been isolated from West Africans. When compared to HIV-1, the nucleotide sequence, in vitro properties, and clinical behavior of HIV-2 define it as a related but unique virus. HIV-1 is a cause of acquired immunodeficiency syndrome (AIDS), wasting, neurologic syndromes, and death in much of the world. HIV-2 may be less pathogenic. This is a proposal to study structural and functional properties of the envelope glycoproteins of HIV-2, answering questions important to the biology, pathogenesis, immunology, and therapy of all HIV infection. The particular emphasis on HIV-2 envelope glycoproteins and on HIV-induced syncytium formation in CD4-positive lymphocytes reflects: 1) the proposed major role of HIV-induced syncytia in lymphocyte depletion and subsequent AIDS; 2) the report of an isolate of HIV-2 which does not cause syncytium formation and which is comprised of a mixture of viruses with full-length and truncated transmembrane glycoproteins; and 3) preliminary reports of a reduced incidence of AIDS in West Africans infected with HIV-2 when compared to HIV-1 infected patients elsewhere. The specific objectives are to: 1) express the envelope gene of known and novel isolates of HIV-2 in eukaryotic cells using a recombinant vaccinia virus system; employ the recombinants in the experiments below; make polyclonal rabbit antisera to the recombinant HIV-2 glycoproteins; 2) study fusion activity of different HIV-2 isolates, also characterizing the pH optimum; host cell dependence of cleavage; effects of potential fusion inhibitors; and possible existence of a host cell fusion receptor distinct from the binding receptor; 3) predict, through deduced amino acid sequence comparison, the crucial molecular features in HIV-2 isolates observed to lack fusion, and test these predictions by site-directed mutagenesis studies; 4) study the biologic results of naturally occurring and experimental truncation of the transmembrane glycoprotein of HIV-2 specifically evaluating fusion, oligomerization, polarized expression, lateral mobility, and assembly into virions; and 5) develop conditions for large scale production and crystallization of HIV-2 envelope glycoproteins. The network of information and materials, the physical proximity of the laboratories of the investigators, and the complementary research projects encourage innovative collaboration in this PEBRA. As indicated in the proposal, this project will involve extensive collaborative efforts with other PEBRA investigators.

人类免疫缺陷病毒 2 (HIV-2) 已从 西非人。 与 HIV-1 相比，核苷酸 HIV-2 的序列、体外特性和临床行为 将其定义为一种相关但独特的病毒。 HIV-1 是导致 获得性免疫缺陷综合症（艾滋病）、消瘦、神经系统 世界大部分地区的综合症和死亡。 HIV-2可能较少 致病的。 这是研究结构和功能的提案 HIV-2 包膜糖蛋白的特性，回答 对生物学、发病机制、免疫学等重要的问题 治疗所有艾滋病毒感染。 特别强调 HIV-2 包膜糖蛋白和 HIV 诱导的合胞体形成 在CD4阳性淋巴细胞中反映：1）所提出的主要作用 HIV诱导的合胞体在淋巴细胞耗竭和随后的过程中的作用 艾滋病; 2) HIV-2 分离株的报告，该分离株不会引起 合胞体形成，它由以下物质的混合物组成： 具有全长和截短跨膜的病毒 糖蛋白； 3) 发病率降低的初步报告 与其他国家相比，感染 HIV-2 的西非人患艾滋病的比例 其他地方感染了 HIV-1 的患者。 具体目标是： 1) 表达已知和新的HIV-2分离株的包膜基因 在真核细胞中使用重组痘苗病毒系统； 在下面的实验中使用重组体；制作 重组HIV-2糖蛋白的多克隆兔抗血清； 2) 研究不同HIV-2分离株的融合活性，同时 表征最佳 pH 值；裂解的宿主细胞依赖性； 潜在融合抑制剂的影响；以及可能存在的 宿主细胞融合受体不同于结合受体； 3） 通过推论氨基酸序列比较，预测 观察到 HIV-2 分离株缺乏关键分子特征 融合，并通过定点突变测试这些预测 研究； 4）研究自然发生的生物学结果 跨膜糖蛋白的实验截断 HIV-2 专门评估融合、寡聚、极化 表达、横向移动性和组装成病毒体；和 5) 开发大规模生产和结晶的条件 HIV-2 包膜糖蛋白。 信息网络和 材料、实验室的物理距离 研究人员和补充研究项目鼓励 PEBRA 中的创新合作。 如中所示 建议，该项目将涉及广泛的协作努力 与其他 PEBRA 调查员一起。