Mannosidase-stabilized glycan immunogens for elicitation of high mannose patch antibodies

甘露糖苷酶稳定的聚糖免疫原，用于引发高甘露糖贴片抗体

• 批准号: 10619737
• 负责人: Isaac Jonathan Krauss
• 金额: $ 24.38万
• 依托单位: BRANDEIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10619737
• 关键词: Affinity; Antibodies; Antibody Response; Antigens; Binding; Carbohydrates; Carrier Proteins; Collaborations; Disaccharides; Enzyme-Linked Immunosorbent Assay; Epitopes; Family; Family member; Glycoconjugates; Glycopeptides; Goals; HIV; HIV Antibodies; HIV Envelope Protein gp120; HIV Infections; Hydrophobicity; Immunization; Immunize; Infection; Laboratories; Length; Macaca mulatta; Mannose; Mannosidase; Memory B-Lymphocyte; Methods; Modification; Oryctolagus cuniculus; Oxygen; Patients; Peptides; Polysaccharides; Proteins; Serum; Structure; Structure of germinal center of lymph node; Sulfur; Surface; Testing; V3 Loop; Vaccination; Vaccines; Variant; Virus; X-Ray Crystallography; analog; arm; cross reacting material 197; glycosylation; immunogenicity; improved; in vivo; man; mannosyl(9)-N-acetylglucosamine2; neutralizing antibody; prevent; response

Project Summary/Abstract HIV broadly neutralizing antibodies (bnAbs) most commonly target the High Mannose Patch (HMP), a mannose-rich region of HIV Env in the vicinity of the V3 loop and the N332 glycan. HMP bnAbs (e.g., 2G12, PGT128, BG18) tend to recognize non-reducing-terminal Man1α-2Man moieties within oligomannose glycans; however, in immunizations with oligomannose glycoconjugates, antibodies are rarely raised against Man1a-2Man, and instead bind to core motifs of the glycan. We hypothesize that in vivo mannosidase trimming of Man1a-2Man termini is a major reason why antibodies do not develop against this motif. In Aim 1, we will prepare stabilized oligomannose glycans, in which a sulfur linkage prevents mannosidase cleavage of the Man1α-2Man, but structurally mimics the natural glycan and is recognized by bnAbs. In Aim 2, we will immunize rabbits with CRM197 glycoconjugates containing the stabilized or unstabilized glycans and compare the resulting antibodies’ HIV Env binding and neutralization activity. We will also intensively characterize the ability of these antibodies to bind Man1a- 2Man using an oligomannose-focused glycan array.

项目总结/摘要 HIV广泛中和抗体（bnAb）最常靶向高甘露糖斑块（HMP）， 在V3环和N332聚糖附近的HIV Env的富含甘露糖的区域。HMP bnAb（例如， 2G 12、PGT 128、BG 18）倾向于识别 寡甘露糖聚糖;然而，在用寡甘露糖糖缀合物的免疫中， 很少针对Man 1a-2 Man产生，而是与聚糖的核心基序结合。我们假设， Man 1a-2 Man末端的体内甘露糖苷酶修剪是抗体不产生的主要原因 反对这个主题。在目标1中，我们将制备稳定的低聚甘露糖聚糖，其中硫键 阻止Man 1 α-2Man的甘露糖苷酶裂解，但在结构上模拟天然聚糖， 被bnAbs识别。在目标2中，我们将用含有CRM 197的糖缀合物免疫兔。 稳定或不稳定的聚糖，并比较所得抗体的HIV Env结合和 中和活性我们还将集中表征这些抗体结合Man 1a的能力， 2 Man使用寡聚甘露糖聚焦聚糖阵列。