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: 9751176
• 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 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9751176
• 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 Investigative Technique; 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.

项目摘要 - 总体 虽然显然需要疫苗引起广泛中和抗体（BNAB）的疫苗 高度可变的逆转录病毒HIV-1可以阻止人与人与全球传播，这是实现这一目标的手段 仍然难以捉摸。天然中脆弱性的三聚体GP160位点产生的抗体 感染进一步推动逆转录病毒突变，使个体中的准特性多样化。一个明显的例外是 隐形的膜近端外部区域（MPER）部位，主要浸入脂质中，仅浸入 在半分解/融合过程中揭示。因此，免疫压力不会混淆或导致病毒 逃脱。但是，已经假定自然产生的抗体很少是 鉴于脂质反应性，多元特异性甚至自动反应性，在最早的B细胞检查点上删除 某些抗污药mab。相反，我们最近的小鼠免疫原性研究产生了生发 中心衍生和T细胞依赖性的，亲和力成熟的抗MPer抗体，使用脂质体阵列MPER 没有临床后遗症或明显的抗脂反应性的片段。这些发现与重新 评估了HIV-1患者中人类抗MPER抗体的自然出现的较高患病率。这个po1 专注于通过有效的BNAB特异性发挥反MPER反应，从而产生免疫原分 捕获本地MPER构象状态在三聚体过渡期间产生 紧凑的圆环状GP160碱基，以最大的MPer暴露在平坦的病毒膜上。这 应使用Reinherz组（DFCI）项目1中的分子免疫学方法来实现目标 与项目2中尔湾集团（MIT）的生物材料专业知识结合并利用技术 组件A-D，NMR（Wagner，Harvard），电子显微镜（EM）（Walz，Rockefeller），EPR（歌曲，， NHMFL）和微磨碎（爱，麻省理工学院）。免疫原设计的优化将通过 通过使用合成肽核酸，MPER的方向在纳米酮中的脂质膜上取向 （PNA）茎，酰基链和TM段接头，并由NMR，EPR，EM和BIACORE测量审查。在 此外，最佳的纳米级MPER细分段和组织将参与不合时宜的共同点 BNABS的祖先（UCA）。抗体附着的矢量空间将由生物材料配方控制 聚合物聚（乙二醇）（PEG）的“空间云”和纳米盘填充的GP160的利用率 异源免疫策略。通过单细胞微磨格挽救的引发mab的接近角度 将评估并将其与自然产生的bnabs进行比较。化学佐剂的共同传递 激活STING和ICOS途径应在生发中心增加CD4 TFH以驱动体细胞 超名。通过分析散装血清评估疫苗诱导的长期浆细胞发育 使用常规小鼠以及完整的Ig- 基因座人性化小鼠（Kymouse）将允许在相关的临床前模型系统中进行免疫调节。