Discovery and characterization of SARS-CoV-2 reactive and neutralizing antibodies from humanized CAMouse(HG) mice through rapid hybridoma screening and high-throughput single-cell V(D)J sequencing.
Discovery and characterization of SARS-CoV-2 reactive and neutralizing antibodies from humanized CAMouse(HG) mice through rapid hybridoma screening and high-throughput single-cell V(D)J sequencing.
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DOI：
10.3389/fimmu.2022.992787
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发表时间：
2022
影响因子：
7.3
通讯作者：
Ge, Liangpeng
中科院分区：
文献类型：
--
作者：
Yang, Xi;Chi, Hang;Wu, Meng;Wang, Zhenshan;Lang, Qiaoli;Han, Qiuxue;Wang, Xinyue;Liu, Xueqin;Li, Yuanguo;Wang, Xiwen;Huang, Nan;Bi, Jinhao;Liang, Hao;Gao, Yuwei;Zhao, Yongkun;Feng, Na;Yang, Songtao;Wang, Tiecheng;Xia, Xianzhu;Ge, Liangpeng
关键词：
到目前为止，冠状病毒2019年大流行已造成5.32亿多人感染，630万人死亡。反应性和中和性的SARS-CoV-2全人抗体是有效的检测工具和治疗措施。在SARS-CoV-2感染过程中，会产生大量的SARS-CoV-2反应性抗体和中和抗体。大多数SARS-CoV-2反应性和中和性全人类抗体是从人类中分离出来的，通常由收敛的重链可变基因编码。然而，SARS-CoV-2病毒可以在复制过程中迅速变异，中和抗体的抗药性变种很容易存活并逃避免疫反应，特别是在面对人类这种集中的抗体反应时。因此，需要更多的工具来开发不同类型的完全人类抗体来弥补目前的不足。在本研究中，我们利用抗体人源化的CAMouseHG小鼠建立了一种快速的抗体发现方法，并通过高通量单细胞V(D)J测序分析检测了CAMouseHG小鼠来源的SARS-CoV-2 RBD反应性杂交瘤细胞的抗体库。用28d快速免疫法免疫CAMouseHG小鼠。经电融合和电融合后第12天的半固体培养筛选，根据SARS-CoV-2 RBD结合活性测定结果，获得171个杂交瘤克隆。在CAMouseHG小鼠杂交瘤克隆中发现IGHV6-1家族具有相当明显的偏好作用，这与新冠肺炎患者中发现的抗体有显著差异。经过进一步的病毒中和筛选和抗体竞争试验，我们获得了一种非竞争性双抗体鸡尾酒，该鸡尾酒对食蟹猴SARS-CoV-2具有很强的预防保护作用。这些结果表明，人源化的CAMouseHG小鼠不仅为获得完全的人反应性和中和性抗体提供了一个有价值的平台，而且具有不同于人类的抗体谱。因此，人源化CAMouseHG小鼠可以作为发现完全人类治疗性和诊断性抗体的良好补充工具。
The coronavirus disease 2019 pandemic has caused more than 532 million infections and 6.3 million deaths to date. The reactive and neutralizing fully human antibodies of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are effective detection tools and therapeutic measures. During SARS-CoV-2 infection, a large number of SARS-CoV-2 reactive and neutralizing antibodies will be produced. Most SARS-CoV-2 reactive and neutralizing fully human antibodies are isolated from human and frequently encoded by convergent heavy-chain variable genes. However, SARS-CoV-2 viruses can mutate rapidly during replication and the resistant variants of neutralizing antibodies easily survive and evade the immune response, especially in the face of such focused antibody responses in humans. Therefore, additional tools are needed to develop different kinds of fully human antibodies to compensate for current deficiency. In this study, we utilized antibody humanized CAMouseHG mice to develop a rapid antibody discovery method and examine the antibody repertoire of SARS-CoV-2 RBD-reactive hybridoma cells derived from CAMouseHG mice by using high-throughput single-cell V(D)J sequencing analysis. CAMouseHG mice were immunized by 28-day rapid immunization method. After electrofusion and semi-solid medium screening on day 12 post-electrofusion, 171 hybridoma clones were generated based on the results of SARS-CoV-2 RBD binding activity assay. A rather obvious preferential usage of IGHV6-1 family was found in these hybridoma clones derived from CAMouseHG mice, which was significantly different from the antibodies found in patients with COVID-19. After further virus neutralization screening and antibody competition assays, we generated a noncompeting two-antibody cocktail, which showed a potent prophylactic protective efficacy against SARS-CoV-2 in cynomolgus macaques. These results indicate that humanized CAMouseHG mice not only provide a valuable platform to obtain fully human reactive and neutralizing antibodies but also have a different antibody repertoire from humans. Thus, humanized CAMouseHG mice can be used as a good complementary tool in discovery of fully human therapeutic and diagnostic antibodies.
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DOI：
10.1056/nejmoa2029849
发表时间：
2021-01-21
期刊：
The New England journal of medicine
影响因子：
--
作者：
Chen P;Nirula A;Heller B;Gottlieb RL;Boscia J;Morris J;Huhn G;Cardona J;Mocherla B;Stosor V;Shawa I;Adams AC;Van Naarden J;Custer KL;Shen L;Durante M;Oakley G;Schade AE;Sabo J;Patel DR;Klekotka P;Skovronsky DM;BLAZE-1 Investigators
通讯作者：
BLAZE-1 Investigators
DOI：
10.1056/nejmoa2102685
发表时间：
2021-10-07
期刊：
The New England journal of medicine
影响因子：
--
作者：
Dougan M;Nirula A;Azizad M;Mocherla B;Gottlieb RL;Chen P;Hebert C;Perry R;Boscia J;Heller B;Morris J;Crystal C;Igbinadolor A;Huhn G;Cardona J;Shawa I;Kumar P;Adams AC;Van Naarden J;Custer KL;Durante M;Oakley G;Schade AE;Holzer TR;Ebert PJ;Higgs RE;Kallewaard NL;Sabo J;Patel DR;Dabora MC;Klekotka P;Shen L;Skovronsky DM;BLAZE-1 Investigators
通讯作者：
BLAZE-1 Investigators
影响因子：
9.5
作者：
Jin, Xiyun;Zhou, Wenyang;Jiang, Qinghua
通讯作者：
Jiang, Qinghua
影响因子：
64.5
作者：
Corti D;Purcell LA;Snell G;Veesler D
通讯作者：
Veesler D
影响因子：
32.4
作者：
Schultheiss, Christoph;Paschold, Lisa;Binder, Mascha
通讯作者：
Binder, Mascha