Complete humanization of the mouse immunoglobulin loci enables efficient therapeutic antibody discovery
Complete humanization of the mouse immunoglobulin loci enables efficient therapeutic antibody discovery
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DOI：
10.1038/nbt.2825
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发表时间：
2014-04-01
影响因子：
46.9
通讯作者：
Bradley, Allan
中科院分区：
文献类型：
--
作者：
Lee, E-Chiang;Liang, Qi;Bradley, Allan
如果用感兴趣的抗原免疫，具有大部分未重排的人免疫球蛋白基因座的转基因小鼠可以产生完全人抗原特异性抗体;几种这样的抗体在临床上使用。然而，常规转基因技术固有的技术限制和人与小鼠免疫球蛋白恒定区之间的序列差异限制了这些小鼠的实用性。在这里，使用胚胎干细胞中基因组工程的重复循环，我们已经将整个人免疫球蛋白可变基因库（2.7 Mb）插入小鼠基因组中，使小鼠恒定区保持完整。这些转基因小鼠是可行的和可繁殖的，具有类似于野生型小鼠的免疫系统。抗原免疫导致产生具有长的人样互补决定区3（CDR 3 H）、宽的表位覆盖和体细胞超突变的强特征的高亲和力抗体。这些小鼠为发现治疗性人类单克隆抗体提供了一个强大的系统;作为人类抗体反应的替代读数，它们也可以帮助疫苗设计工作。
If immunized with an antigen of interest, transgenic mice with large portions of unrearranged human immunoglobulin loci can produce fully human antigen-specific antibodies; several such antibodies are in clinical use. However, technical limitations inherent to conventional transgenic technology and sequence divergence between the human and mouse immunoglobulin constant regions limit the utility of these mice. Here, using repetitive cycles of genome engineering in embryonic stem cells, we have inserted the entire human immunoglobulin variable-gene repertoire (2.7 Mb) into the mouse genome, leaving the mouse constant regions intact. These transgenic mice are viable and fertile, with an immune system resembling that of wild-type mice. Antigen immunization results in production of high-affinity antibodies with long human-like complementarity-determining region 3 (CDR3H), broad epitope coverage and strong signatures of somatic hypermutation. These mice provide a robust system for the discovery of therapeutic human monoclonal antibodies; as a surrogate readout of the human antibody response, they may also aid vaccine design efforts.