Targeting Immunogenicity to the MPER Hinge and C-helix for BNAb Elicitation

将免疫原性靶向 MPER 铰链和 C 螺旋以诱导 BNAb

• 批准号: 9982752
• 负责人: ELLIS L REINHERZ
• 金额: $ 174.47万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-05 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9982752
• 关键词: Adjuvant; Affinity; Albumins; Amino Acid Sequence; Antibodies; Antibody Affinity; Antibody Repertoire; Antibody Response; Antibody Specificity; Antigens; B-Lymphocytes; Binding; Biocompatible Materials; Biological Models; Biosensor; Bone Marrow; Cells; Chemicals; Chemistry; Clinical; Cohort Studies; Conserved Sequence; Cryoelectron Microscopy; Development; Dinucleoside Phosphates; Electron Microscopy; Failure; Formulation; Frequencies; Future; Geometry; Goals; HIV Envelope Protein gp120; HIV-1; Health; Human; Immersion; Immune; Immunization; Immunoglobulin G; Immunoglobulin Somatic Hypermutation; Individual; Infection; Link; Lipid Binding; Lipids; Liposomes; Love; Measurement; Mediating; Membrane; Membrane Lipids; Methods; Modification; Molecular Conformation; Molecular Immunology; Monoclonal Antibodies; Mus; Mutate; Mutation; Nature; Neutralization Tests; Pathway interactions; Patients; Peptide Nucleic Acids; Periodicity; Persons; Plasma Cells; Polymers; Polysaccharides; Pre-Clinical Model; Prevalence; Preventive; Preventive vaccine; Reaction; Retroviridae; Serum; Signal Transduction; Site; Specificity; Structure; Structure of germinal center of lymph node; T-Lymphocyte; Technology; United States National Institutes of Health; Vaccination; Vaccines; Variant; Viral; Virion; adduct; autoreactivity; base; biophysical properties; conformer; density; design; ethylene glycol; global health; gp160; humanized mouse; immunogenicity; lymph nodes; mutant; nanodisk; nanoscale; nanovaccine; neutralizing antibody; plasma cell development; pressure; receptor binding; response; stem; stereochemistry; synthetic peptide; vaccine development; vector

Project Summary - Overall While there is an obvious need for vaccines eliciting broadly neutralizing antibodies (BNAbs) against the highly mutable retrovirus HIV-1 to stem person-to-person and global spread, the means to achieve this goal have remained elusive. Antibodies produced against trimeric gp160 sites of vulnerability during natural infection drive retroviral mutation further, diversifying quasispecies in individuals. One apparent exception is the membrane proximal external region (MPER) site, which is stealth, largely immersed in lipid and only revealed during hemifusion/fusion. Hence, immune pressures are not confounding or contributing to viral escape. However, a paucity of naturally arising antibodies has been postulated to be a consequence of deletion at the earliest B cell checkpoints in view of lipid reactivity, polyspecificity and even autoreactivity of certain anti-MPER mAbs. To the contrary, our recent mouse immunogenicity studies have generated germinal center-derived and T cell-dependent, affinity-matured anti-MPER antibodies using liposome-arrayed MPER segments without clinical sequelae or significant anti-lipid reactivity. These findings are consistent with the re- evaluated greater prevalence of naturally arising human anti-MPER antibodies in HIV-1 patients. This PO1 focuses on engendering anti-MPER responses with potent BNAb specificities, creating immunogens that capture native MPER conformational states arising during transitions of trimer unwinding, going from the compact torus-like gp160 base to conformers on the flat viral membrane with maximum MPER exposure. This goal shall be achieved using molecular immunology approaches of the Reinherz group (DFCI) in Project 1 in conjunction with the biomaterials expertise of the Irvine group (MIT) in Project 2 and utilizing technology components A-D, NMR (Wagner, Harvard), electron microscopy (EM) (Walz, Rockefeller), EPR (Song, NHMFL) and microengraving (Love, MIT), respectively. Optimization of immunogen design will be achieved by orientation of the MPER on lipid membranes in nanovaccines through the use of synthetic peptide nucleic acid (PNA) stalks, acyl chain and TM segment linkers and vetted by NMR, EPR, EM and BIAcore measurements. In addition, optimal nanoscale MPER segment spacing and organization will engage uncommitted common ancestors (UCAs) of BNAbs. Vector space of antibody attachment will be controlled by biomaterial formulation of polymer poly(ethylene glycol) (PEG) "steric clouds" and utilization of nanodisc-embedded gp160 in heterologous immunization strategies. Approach angles of elicited mAbs rescued by single-cell microengraving will be assessed and compared with those of naturally arising BNAbs. Co-delivery of chemical adjuvants to activate the STING and ICOS pathways shall augment CD4 TFH in germinal centers to drive somatic hypermutation. Assessment of vaccine-induced long-term plasma cell development by analysis of bulk serum IgG and single bone marrow (BM) plasma cell microengraving using conventional mice as well as complete Ig- locus humanized mice (KyMouse) will allow for immunogen tuning in a relevant pre-clinical model system.

项目总结 - 总体 虽然显然需要能够引发广泛中和抗体（BNAb）的疫苗 高度可变的逆转录病毒 HIV-1 可以阻止人际传播和全球传播，这是实现这一目标的手段 仍然难以捉摸。自然过程中针对三聚体 gp160 脆弱位点产生的抗体 感染进一步推动逆转录病毒突变，使个体准种多样化。一个明显的例外是 膜近端外部区域（MPER）位点，它是隐形的，大部分浸入脂质中，并且仅 在半融合/融合期间显示。因此，免疫压力不会混淆或促进病毒传播 逃脱。然而，天然产生的抗体的缺乏被认为是以下因素的结果： 鉴于脂质反应性、多特异性甚至自身反应性，在最早的 B 细胞检查点进行删除 某些抗 MPER 单克隆抗体。相反，我们最近的小鼠免疫原性研究已经产生了生发 使用脂质体阵列 MPER 获得中心衍生的 T 细胞依赖性、亲和力成熟的抗 MPER 抗体 没有临床后遗症或显着抗脂质反应的片段。这些发现与重新研究一致 评估了 HIV-1 患者中自然产生的人类抗 MPER 抗体的流行率。这个PO1 专注于产生具有有效 BNAb 特异性的抗 MPER 反应，创造免疫原 捕获三聚体解旋转变期间产生的天然 MPER 构象状态，从 紧凑的环面状 gp160 碱基与扁平病毒膜上的构象异构体具有最大的 MPER 暴露。这 目标应使用Reinherz小组（DFCI）的分子免疫学方法在项目1中实现 项目 2 中结合欧文集团 (MIT) 的生物材料专业知识并利用技术 成分 A-D、NMR（瓦格纳、哈佛）、电子显微镜 (EM)（沃尔兹、洛克菲勒）、EPR（宋、 NHMFL）和微雕刻（Love，麻省理工学院）。免疫原设计的优化将通过以下方式实现 通过使用合成肽核酸在纳米疫苗中的脂质膜上定位 MPER (PNA) 茎、酰基链和 TM 段接头，并通过 NMR、EPR、EM 和 BIAcore 测量进行审查。在 此外，最佳的纳米级 MPER 段间距和组织将涉及未承诺的共同点 BNAb 的祖先 (UCA)。抗体附着的载体空间将由生物材料配方控制 聚合物聚乙二醇（PEG）“空间云”的制备以及纳米盘嵌入的 gp160 在 异源免疫策略。通过单细胞微雕刻拯救引发的单克隆抗体的接近角度 将进行评估并与自然产生的 BNAb 进行比较。将化学助剂共同递送至 激活 STING 和 ICOS 途径将增强生发中心的 CD4 TFH 以驱动体细胞 超突变。通过分析大量血清评估疫苗诱导的长期浆细胞发育 使用传统小鼠以及完整 Ig- 进行 IgG 和单骨髓 (BM) 浆细胞微雕刻 位点人源化小鼠（KyMouse）将允许在相关的临床前模型系统中进行免疫原调整。

项目成果

Inadequate structural constraint on Fab approach rather than paratope elicitation limits HIV-1 MPER vaccine utility.

• DOI: 10.1038/s41467-023-42097-6
• 发表时间: 2023-11-08
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Tan K;Chen J;Kaku Y;Wang Y;Donius L;Khan RA;Li X;Richter H;Seaman MS;Walz T;Hwang W;Reinherz EL;Kim M
• 通讯作者: Kim M

Dynamic HIV-1 spike motion creates vulnerability for its membrane-bound tripod to antibody attack.

• DOI: 10.1038/s41467-022-34008-y
• 发表时间: 2022-10-27
• 期刊: Nature communications
• 影响因子: 16.6