Protective Human Monoclonal Antibodies to HIV1

HIV1 保护性人单克隆抗体

• 批准号: 7237936
• 负责人: Susan B Zolla-Pazner
• 金额: $ 65.24万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-05-01 至 2008-08-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7237936
• 关键词: Antibodies; Antibody Binding Sites; Antigens; Binding; Binding Sites; Biochemical; Biological Assay; Categories; Cells; Chemicals; Codon Nucleotides; Complement; Complementarity Determining Regions; Complex; Data; Development; Directed Molecular Evolution; Dissection; Engineering; Epitope Mapping; Epitopes; Excision; Glycoproteins; HIV; HIV Infections; HIV vaccine; HIV-1; Human; Immunologics; Individual; Libraries; Luciferases; Measures; Methods; Modification; Molecular; Mutagenesis; Nature; Numbers; Parents; Passive Immunization; Peptide Phage Display Library; Peptides; Phage Display; Play; Proteins; Random Peptide Libraries; Reagent; Resistance; Roentgen Rays; Role; Screening procedure; Serum; Structural Protein; Structure; Sum; Techniques; Technology; Testing; Vaccines; Virion; Virus; Virus Diseases; arginyllysine; base; cross reactivity; design; env Genes; human monoclonal antibodies; improved; intermolecular interaction; neutralizing antibody; neutralizing monoclonal antibodies; prophylactic; research study; virus identification

DESCRIPTION (provided by applicant): Currently available human monoclonal antibodies (mAbs) which neutralize primary isolates of HIV are directed against only a few regions of the HIV envelope glycoproteins. Some primary isolates are resistant to many of these mAbs but can be neutralized by pooled sera from HIV-infected individuals, suggesting that there may be additional neutralizing epitopes which have not yet been identified. To develop new mAbs with broad and potent neutralizing activity whose activity will complement those of previously described mAbs, which can be used for passive immunization, and whose epitopes can be characterized and incorporated into the design of candidate HIV vaccines, we propose to: (a) Produce human mAbs with potent and broad neutralizing activity against HIV-1 primary isolates that target previously unidentified epitopes. Existing human anti-HIV mAbs have been selected on the basis of their binding activity. We propose to select anti-HIV mAbs based on their function, i.e., their ability to neutralize pseudovirions containing env genes of various primary isolates. (b) Delineate the breadth of the neutralizing activity of the new mAbs selected with the neutralization screen. (c) Use a variety of immunologic, biochemical, molecular and physical techniques to identify and characterize the epitopes recognized by the broadest and most potent of the mAbs. Epitope mapping techniques will include the use of an antigen fragment library, random peptide phage display libraries, epitope excision/extraction and Arg/Lys chemical modification methods, and X-ray crystallographic analyses. These studies will elucidate the nature of the epitopes recognized by the best of the neutralizing mAbs, providing critical information about the structure of epitopes that induce protective Abs. These data will also elucidate the structure of the mAb paratopes, the intermolecular interactions between epitopes and paratopes, and the molecular basis for the cross-reactivity between diverse strains of HIV. (d) Use a "focused mutagenesis" technique to engineer two selected mAbs in order to improve their potency and breadth, and subsequently determine the structure of the epitopes recognized by these engineered mAbs. The results of these studies should provide both reagents for passive immunization and data on the nature of antigenic determinants of HIV glycoproteins useful for designing immunogens that will induce broadly neutralizing Abs.

描述（由申请人提供）：目前可用的中和HIV初级分离株的人单克隆抗体（mAb）仅针对HIV包膜糖蛋白的少数区域。 一些初级分离株对许多单克隆抗体具有抗性，但可以被 HIV 感染者的混合血清中和，这表明可能存在其他尚未确定的中和表位。 为了开发具有广泛和有效的中和活性的新单克隆抗体，其活性将补充先前描述的可用于被动免疫的单克隆抗体，并且其表位可以被表征并纳入候选HIV疫苗的设计中，我们建议：（a）生产针对针对先前未识别的表位的针对HIV-1初级分离株具有有效和广泛的中和活性的人单克隆抗体。 现有的人类抗 HIV 单克隆抗体是根据其结合活性进行选择的。 我们建议根据抗 HIV 单克隆抗体的功能来选择，即它们中和含有各种初级分离株的 env 基因的假病毒粒子的能力。 (b) 描绘通过中和筛选选择的新单克隆抗体的中和活性的广度。 (c) 使用各种免疫学、生物化学、分子和物理技术来鉴定和表征最广泛和最有效的单克隆抗体所识别的表位。 表位作图技术将包括使用抗原片段库、随机肽噬菌体展示库、表位切除/提取和Arg/Lys化学修饰方法以及X射线晶体学分析。 这些研究将阐明最好的中和单克隆抗体所识别的表位的性质，提供有关诱导保护性抗体的表位结构的关键信息。 这些数据还将阐明 mAb 互补位的结构、表位和互补位之间的分子间相互作用，以及不同 HIV 毒株之间交叉反应的分子基础。 (d) 使用“集中诱变”技术对两种选定的单克隆抗体进行工程改造，以提高其效力和广度，并随后确定这些工程单克隆抗体识别的表位的结构。 这些研究的结果应该提供用于被动免疫的试剂和关于HIV糖蛋白抗原决定簇性质的数据，这些数据可用于设计可诱导广泛中和抗体的免疫原。