Protective Human Monoclonal Antibodies to HIV-1

HIV-1 保护性人单克隆抗体

• 批准号: 8759759
• 负责人: Susan B Zolla-Pazner
• 金额: $ 79.96万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-12-01 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8759759
• 关键词: Active Immunization; Anti-Retroviral Agents; Antibodies; Area; Binding; Binding Sites; Biological Assay; CCR5 gene; Cameroon; Cell fusion; Cells; Characteristics; Complement; Complex; Derivation procedure; Development; Enzyme-Linked Immunosorbent Assay; Epitope Mapping; Epitopes; Fc Receptor; Foundations; Funding; Generations; Grant; HIV; HIV Antibodies; HIV Envelope Protein gp120; HIV Infections; HIV vaccine; HIV-1; Health; Human; Hybridomas; Immunization; Immunologics; Immunotherapy; Individual; Infection; Knowledge; Lead; Mannose; Maps; Measures; Mediating; Membrane; Methods; Nature; Participant; Passive Immunization; Pathway interactions; Patients; Pattern; Peripheral Blood Mononuclear Cell; Pharmaceutical Preparations; Population; Prevention; Production; Radiolabeled; Reagent; Resistance; Serum; Specificity; Structure; Techniques; Testing; Toxin; Tube; United States National Institutes of Health; V3 Loop; Vaccine Design; Vaccines; Viral; Virus; Virus Replication; Work; base; cell transformation; cross reactivity; design; drug development; human monoclonal antibodies; insight; mutant; neutralizing antibody; novel; prevent; radiotracer; screening

DESCRIPTION (provided by applicant): Attempts to induce broadly reactive and potent protective anti-HIV antibodies (Abs) by immunization of humans have been unsuccessful. The optimal vaccine-induced Abs are usually considered to be "neutralizing Abs" that have been defined by studies of several human monoclonal Abs (mAbs) as targeting epitopes in the membrane proximal external region of gp41 and in various regions of gp120 including the CD4 binding site, a complex of high mannose residues, the bridging sheet, and the V3 loop. However, HIV-infected individuals are able to produce both neutralizing Abs that target additional epitopes and "non-neutralizing" Abs that block virus replication through pathways other than that mediated by pH-independent virus/cell fusion. These latter "non-neutralizing" Abs inhibit viral replication through a variety of mechanisms such as those mediated by Fc receptor-bearing cells and by complement. Based on these findings, we propose to focus efforts on identifying new protective human mAbs, i.e., both "conventional" neutralizing mAbs specific for previously unrecognized envelope (Env) epitopes, and human mAbs that inhibit virus replication via "non-neutralizing" pathways. For this, hybridomas will be generated from cells of US and Cameroon subjects infected with HIV strains carrying clade B and A Envs, respectively. Three new screening methods will be used for mAb selection, and a variety of methods will be employed to test the ability of the new mAbs to inhibit virus replication. The work is divided into three aims: 1) Identification of new neutralizing mAbs which target previously unrecognized epitopes, and selection of mAbs with virus inhibitory activities that function through "non-neutralizing" pathways. Pseudoviruses, Env-expressing cells and gp120 reagents will be used in several screening assays, each chosen to maximize the different types and cross-reactivity of mAbs to be selected, and to determine if there are epitopes that are uniquely or preferentially targeted by newly transmitted viruses. 2) Delineation of the inhibitory activities of these mAbs, of their breadth against viruses and pseudoviruses from multiple clades, and of the different patterns of reactivity between the clade B- and A- derived mAbs. These comparisons, together with descriptions of the immunologic nature and specificities of the mAbs, will provide the first comprehensive and systematic information about the characteristics of mAbs with different protective activities. And, 3) Mapping of the epitopes targeted by the new mAbs. Immunologic, virologic and physicochemical techniques will be used to delineate mAb epitopes, the specific interactions between mAb and Env, and quaternary Env structures targeted by mAbs. The reagents generated and the knowledge gained will illuminate mechanisms of virus neutralization and pathways that interrupt virus replication, and will inform the design of reagents for use in active immunization (vaccines), passive immunization, and immunotherapy.

描述（由申请人提供）：通过人类免疫诱导广泛反应性和强效保护性抗hiv抗体（Abs）的尝试尚未成功。最佳的疫苗诱导抗体通常被认为是“中和抗体”，通过对几种人单克隆抗体（mab）的研究，这些抗体被定义为靶向gp41的膜近端外区和gp120的各个区域的表位，包括CD4结合位点、高甘糖残基复合物、桥接片和V3环。然而，hiv感染的个体既能够产生靶向额外表位的中和抗体，也能够产生通过不依赖ph的病毒/细胞融合介导的途径阻断病毒复制的“非中和”抗体。后一种“非中和”抗体通过多种机制抑制病毒复制，如由Fc受体承载细胞和补体介导的机制。基于这些发现，我们建议将重点工作集中在鉴定新的保护性人类单抗上，即“传统”的针对先前未被识别的包膜（Env）表位特异性的中和单抗，以及通过“非中和”途径抑制病毒复制的人单抗。为此，杂交瘤将从分别携带进化支B和A Envs的HIV株感染的美国和喀麦隆受试者的细胞中产生。将采用三种新的筛选方法进行单抗的筛选，并采用多种方法测试新单抗抑制病毒复制的能力。这项工作分为三个目标：1)鉴定新的靶向以前未被识别的表位的中和单抗，以及选择通过“非中和”途径发挥作用的具有病毒抑制活性的单抗。假病毒、表达env的细胞和gp120试剂将用于几种筛选试验，每种选择都是为了最大限度地提高所选单克隆抗体的不同类型和交叉反应性，并确定是否存在新传播病毒唯一或优先靶向的表位。2)描述了这些单抗的抑制活性，它们对来自多个分支的病毒和假病毒的抑制广度，以及B和A分支衍生的单抗的不同反应模式。这些比较，连同对单克隆抗体的免疫学性质和特异性的描述，将提供关于具有不同保护活性的单克隆抗体特征的第一个全面和系统的信息。3)新单克隆抗体靶向的抗原表位定位。免疫学、病毒学和物理化学技术将用于描述单抗表位、单抗与Env之间的特异性相互作用以及单抗靶向的四级Env结构。生成的试剂和获得的知识将阐明病毒中和的机制和中断病毒复制的途径，并将为设计用于主动免疫（疫苗）、被动免疫和免疫治疗的试剂提供信息。

项目成果

Thermodynamic signatures of the antigen binding site of mAb 447-52D targeting the third variable region of HIV-1 gp120.

• DOI: 10.1021/bi400645e
• 发表时间: 2013-09-10
• 期刊: BIOCHEMISTRY
• 影响因子: 2.9
• 作者: Killikelly, April;Zhang, Hui-Tang;Spurrier, Brett;Williams, Constance;Gorny, Miroslaw K.;Zolla-Pazner, Susan;Kong, Xiang-Peng
• 通讯作者: Kong, Xiang-Peng

Epitope mapping of conformational V2-specific anti-HIV human monoclonal antibodies reveals an immunodominant site in V2.

• DOI: 10.1371/journal.pone.0070859
• 发表时间: 2013
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Mayr LM;Cohen S;Spurrier B;Kong XP;Zolla-Pazner S
• 通讯作者: Zolla-Pazner S

Characterization of a discontinuous epitope of the HIV envelope protein gp120 recognized by a human monoclonal antibody using chemical modification and mass spectrometric analysis.

使用化学修饰和质谱分析对人单克隆抗体识别的 HIV 包膜蛋白 gp120 的不连续表位进行表征。

• DOI: 10.1016/j.jasms.2010.03.031
• 发表时间: 2010
• 期刊: Journal of the American Society for Mass Spectrometry
• 影响因子: 3.2
• 作者: Hager-Braun,Christine;Hochleitner,ElisabethO;Gorny,MiroslawK;Zolla-Pazner,Susan;Bienstock,RachelleJ;Tomer,KennethB
• 通讯作者: Tomer,KennethB

Identification of antibody glycosylation structures that predict monoclonal antibody Fc-effector function.

鉴定预测单克隆抗体FC效应功能的抗体糖基化结构。

• DOI: 10.1097/qad.0000000000000444
• 发表时间: 2014-11-13
• 期刊: AIDS (London, England)
• 作者: Chung AW;Crispin M;Pritchard L;Robinson H;Gorny MK;Yu X;Bailey-Kellogg C;Ackerman ME;Scanlan C;Zolla-Pazner S;Alter G
• 通讯作者: Alter G

Clonal analysis of human anti-V3 monoclonal antibodies selected by a V3 tetramer.

由 V3 四聚体选择的人抗 V3 单克隆抗体的克隆分析。

• DOI: 10.3233/hab-130264
• 发表时间: 2012
• 期刊: Human antibodies
• 作者: Li,Liuzhe;Wang,Xiao-Hong;Banerjee,Sagarika;Volsky,Barbara;Williams,Constance;Moody,MAnthony;Zolla-Pazner,Susan;Gorny,MiroslawK
• 通讯作者: Gorny,MiroslawK