Direct Sequencing of Antibodies of the Influenza Immune Response

流感免疫反应抗体的直接测序

• 批准号: 9409463
• 负责人: Adrian Guthals
• 金额: $ 22.5万
• 依托单位: MAPP BIOPHARMACEUTICAL, INC.
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2018-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9409463
• 关键词: Direct; Sequencing; Antibodies; Influenza; Immune

Abstract The annual epidemics of influenza result in a substantial number of hospitalizations with an estimated 3 to 5 million cases of severe disease, and 300,000 to 500,000 deaths globally. During the 20th century, three major influenza pandemics have occurred with a total mortality of 50–100 million people. Recently, new influenza monoclonal antibodies (mAbs) have been identified that are capable of neutralizing a wide range of viruses. In the last eight years, several broadly neutralizing, stem-reactive antibodies have been identified. Of these stem antibodies, only one recognized all influenza A subtypes however this breadth of recognition was only elicited after in vitro mutagenesis. This suggests that an epitope to guide a universal vaccine strategy has yet be defined. Stem antibodies could provide universal influenza coverage since they are potentially capable of neutralizing both influenza A and B viruses. One strategy to identify neutralizing stem-specific antibodies involves immortalized B cells from immunized individuals. B cell-derived mAbs that cross react with the H5 and H7 HA subtypes have been shown to yield high affinity stem-specific mAbs that can bind to all influenza A HA subtypes. An alternative strategy, that doesn’t involve B cell cloning, starts with an antigen specific population of stem binding polyclonal antibodies (pAbs) that can then be directly sequenced by proteomic tandem mass spectrometry (MS/MS). These pAbs are a preferred source for mAb discovery since the peripheral antibody population, not the peripheral B cell population, is representative of the full humoral immune response. Direct sequencing of peripheral pAbs can rapidly mine this immune diversity to yield mAbs against novel and unique epitopes.

抽象的 每年流感的流行导致大量患者住院治疗， 据估计，全球有 3 至 500 万重症病例，30 万至 50 万例死亡。 20世纪期间，发生了3次重大流感大流行，总死亡人数为 最近，新型流感单克隆抗体 (mAb) 已被用于 50-1 亿人。 在过去的八年里，人们发现了能够中和多种病毒的物质。 已经鉴定出几种广泛中和的干反应性抗体。 抗体，只有一种能够识别所有甲型流感亚型，但是这种识别范围 仅在体外诱变后才被引发，这表明存在指导通用的表位。 疫苗策略尚未确定。干抗体能否提供通用流感疫苗。 覆盖范围，因为它们有可能中和甲型和乙型流感病毒。 鉴定中和干细胞特异性抗体的策略涉及来自以下来源的永生化 B 细胞 与 H5 和 H7 HA 亚型发生交叉反应的 B 细胞衍生的单克隆抗体。 已被证明可产生可与所有甲型流感 HA 结合的高亲和力茎特异性单克隆抗体 另一种不涉及 B 细胞克隆的替代策略是从抗原开始。 特定群体的茎结合多克隆抗体 (pAb)，然后可以直接 通过蛋白质组串联质谱法 (MS/MS) 测序这些 pAb 是优选的。 mAb 发现的来源是外周抗体群体，而不是外周 B 细胞 群体，代表了完整的体液免疫反应。 外周多克隆抗体可以快速挖掘这种免疫多样性，产生针对新颖和独特的单克隆抗体 表位。