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.

摘要 流感的年度流行导致大量的住院治疗， 据估计，全球有300万至500万例严重疾病和30万至50万例死亡。 在20世纪，发生了三次主要的流感大流行， 5000万到1亿人。最近，新的流感单克隆抗体（mAb）已被发现。 鉴定出能够中和多种病毒。在过去的八年里， 已经鉴定了几种广泛中和的茎反应性抗体。这些干 只有一种抗体识别所有甲型流感亚型，但这种识别范围 仅在体外诱变后才被引发。这表明，一个表位，以指导普遍的 疫苗战略尚未确定。干细胞抗体可以提供普遍的流感 因为它们可能能够中和甲型和B型流感病毒。一 鉴定中和B细胞特异性抗体的策略涉及来自 免疫个体。与H5和H7 HA亚型交叉反应的B细胞衍生mAb 已显示产生可结合所有甲型流感HA的高亲和力茎特异性mAb 亚型。另一种不涉及B细胞克隆的策略是从抗原开始 然后可以直接将干细胞结合多克隆抗体（pAb）的特异性群体 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，因为外周抗体群，而不是外周B细胞 群体，代表完整的体液免疫应答。直接测序 外周pAb可以快速挖掘这种免疫多样性，以产生针对新的和独特的 表位