Knock-in mice expressing germline-reverted broadly neutralizing HIV antibodies

表达种系恢复的广泛中和艾滋病毒抗体的敲入小鼠

• 批准号: 10190786
• 负责人: DAVID NEMAZEE
• 金额: $ 67.91万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-05 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10190786
• 关键词: Address; Affinity; Animal Model; Animals; Antibodies; Antibody Binding Sites; Antigens; B-Lymphocytes; Binding; Binding Sites; Cell Count; Cell Line; Cells; Chronic; Clinical Trials; Data; Dose; Engineering; Epitopes; Funding; Gene Mutation; Gene Targeting; Genes; Genetic; Goals; HIV; HIV Antibodies; HIV vaccine; Human; Immune response; Immunization; Immunoglobulin Class Switching; In Vitro; Individual; Infection; Infection Control; Knock-in; Knock-in Mouse; Knowledge; Memory B-Lymphocyte; Modeling; Monkeys; Mus; Mutate; Mutation; Pathway interactions; Patients; Pattern; Peptides; Peripheral Blood Mononuclear Cell; Physiological; Plasma Cells; Polysaccharides; Predictive Value; Primates; Production; Seeds; Site; Somatic Mutation; Sorting - Cell Movement; Specificity; Speed; Structure; Study models; Testing; Vaccination; Vaccine Design; Vaccine Research; Vaccines; Variant; Virus; base; competitive environment; cost; design; engineering design; env Glycoproteins; global health; human model; improved; in vivo; insertion/deletion mutation; insight; mouse model; neutralizing antibody; novel vaccines; optimism; response; simian human immunodeficiency virus; tool; vaccination strategy; vaccine candidate; vaccine trial

This is a renewal R01 application to exploit knock-in mouse models and hypermutating human B cell lines carrying germline-reverted broadly neutralizing antibodies (gl-bNAbs) against HIV Env with the goal to speed HIV vaccine research. Developing an HIV vaccine is a major global health objective as no effective vaccine yet exists. bnAbs to HIV Env glycoprotein can likely control infection but eliciting bnAbs by vaccination is challenging because of Env genetic variability and the fact that responses to conserved Env epitopes are weak. bnAbs are made naturally in some patients, but usually only after years of infection, when they provide no real protection. However, many individual bnAbs can provide passive protection in animal models, suggesting that elicitation of these antibodies could be protective, particularly if bnAbs to several conserved sites can be raised simultaneously. To begin to investigate the best ways to elicit bnAbs by vaccination, we have previously generated B cell lines and knock-in mice carrying inferred gl-bnAbs for VRC01, PGT121, b12, 4E10 and other specificities. gl-bnAb mouse models are valuable for antigen design and vaccination studies because they carry B cells with the potential to become bnAbs after appropriate stimulation and somatic mutation. Placement of the V(D)J genes into the physiological Ig loci allows the knock-in B cells to undergo normal H-chain class switching and V-region hypermutation. Moreover, knock-in mice allow vaccination studies in a convenient, relatively low-cost mouse model. One can seed B cells from these mice at physiologically low numbers in otherwise WT mice and assess their responses in the face of competition from non bnAb clones. In these cell lines and mice, we have been carrying out studies to evaluate whether the imputed gl-bnAbs develop normally and to assess various vaccine candidates and immunization strategies. We have tested so-called germline-targeting immunogens engineered to bind better to gl-bnAbs than immunogens derived from WT Env. In the present proposal we focus on two aspects of vaccine design: the use of validated, rather than inferred, gl-bnAb precursors and an assessment of the ease of selectability of beneficial mutations using somatically-mutating human gl-bnAb cell lines and gl-bnAb mice. We hypothesize that by taking these approaches we will identify preferred gl-bnAbs that most effectively mature to become bnAbs and preferred immunogens or selection paths that should most effectively promote bnAb production when tested in primates. Because the problem of poor precursor affinity to antigen and uncertain paths to affinity maturation by somatic mutation are limitations to all immune responses, knowledge obtained here should be applicable to a variety of vaccine targets.

这是一种续签R01应用程序，用于利用敲门式小鼠模型和超重耗电的人类B细胞系 携带种系的广泛中和抗体（GL-BNABS）针对HIV Env，以速度 HIV疫苗研究。开发艾滋病毒疫苗是全球主要的健康目标，因为没有有效的疫苗 但是存在。 BNABS to HIV ENV糖蛋白可能会控制感染，但通过疫苗引起BNAB是 由于ENV遗传变异性以及对保守的Env表位的反应是挑战性的是 虚弱的。在某些患者中，BNAB自然是生产的，但通常只有在感染了多年后，当它们提供 没有真正的保护。但是，许多单独的BNAB可以在动物模型中提供被动保护， 暗示这些抗体的启发可能具有保护性，尤其是在几个保守的bnabs 站点可以同时提高。为了开始研究通过疫苗引起BNAB的最佳方法，我们 以前已经生成了携带VRC01，PGT121的GL-BNAB的B细胞系和敲入小鼠 B12、4E10和其他特殊性。 GL-BNAB小鼠模型对于抗原设计和疫苗接种很有价值 研究是因为它们携带B细胞具有适当刺激后成为BNAB的潜力 躯体突变。将V（d）J基因放置在生理IG基因座中，使B细胞敲入B细胞 经历正常的H链类切换和V区超名。此外，敲入鼠标允许 在方便，相对低成本的小鼠模型中进行的疫苗接种研究。一个可以从这些小鼠中播种B细胞 在生理学上，否则WT小鼠的数量很少，并在竞争中评估他们的反应 来自非BNAB克隆。在这些细胞系和小鼠中，我们一直在进行研究以评估是否是否 估算的GL-BNAB正常发展并评估各种候选疫苗和免疫 策略。我们已经测试了所谓的靶向种系免疫原，该免疫原为粘合更好地与GL-BNAB结合 比源自WT Env的免疫原子。在本提案中，我们关注疫苗设计的两个方面： 使用经过验证的，而不是推断的GL-BNAB前体，并评估可选性的易度性 使用体体突变​​的人GL-BNAB细胞系和GL-BNAB小鼠的有益突变。我们 假设通过采用这些方法，我们将确定最有效成熟的首选GL-BNAB 成为BNAB和首选免疫原或选择路径，应最有效地促进BNAB 在灵长类动物中测试时产生。因为前体亲和力较差的问题和不确定的问题 通过躯体突变的亲和力成熟的途径是对所有免疫反应的局限性，获得的知识 这里应适用于各种疫苗靶标。