Mice with megabase humanization of their immunoglobulin genes generate antibodies as efficiently as normal mice
Mice with megabase humanization of their immunoglobulin genes generate antibodies as efficiently as normal mice
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DOI：
10.1073/pnas.1324022111
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发表时间：
2014-04-08
影响因子：
11.1
通讯作者：
Yancopoulos, George D.
中科院分区：
文献类型：
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作者：
Murphy, Andrew J.;Macdonald, Lynn E.;Yancopoulos, George D.
经过基因工程改造使其 Ig 基因人源化的小鼠能够在小鼠背景（HumAb 小鼠）内产生人类抗体反应，从而为产生全人类治疗抗体提供了宝贵的平台。不幸的是，现有的 HumAb 小鼠不具备功能齐全的免疫系统，这可能是由于其基因人源化的方式所致。迄今为止，HumAb 小鼠是通过破坏内源性小鼠 Ig 基因并同时在不同的随机位置引入人类 Ig 转基因来产生的。 KO+转基因人源化。正如我们在配套论文中所描述的，我们试图通过使用独特的原位人源化方法，用人类对应物精确替换 6 Mb 的小鼠 Ig 重链和 kappa 轻链可变区种系基因片段，同时保持小鼠恒定区完整，从而使小鼠在体液反应中更有效地使用人类可变区片段。我们推断引入的人类可变区基因片段将在其新的遗传位置中无区别地发挥作用，而保留的小鼠恒定区将允许在小鼠环境中实现最佳的相互作用和所得抗体的选择。我们证明，这些小鼠被称为 VelocImmune 小鼠，因为它们是使用 VelociGene 技术产生的，能够有效产生人：小鼠混合抗体（可快速转化为完全人抗体），并且具有与 WT 小鼠无法区分的功能齐全的体液免疫系统。 10 种不同的全人源抗体快速进入人体临床试验，证实了 VelocImmune 方法的有效性。
Mice genetically engineered to be humanized for their Ig genes allow for human antibody responses within a mouse background (HumAb mice), providing a valuable platform for the generation of fully human therapeutic antibodies. Unfortunately, existing HumAb mice do not have fully functional immune systems, perhaps because of the manner in which their genetic humanization was carried out. Heretofore, HumAb mice have been generated by disrupting the endogenous mouse Ig genes and simultaneously introducing human Ig transgenes at a different and random location; KO-plus-transgenic humanization. As we describe in the companion paper, we attempted to make mice that more efficiently use human variable region segments in their humoral responses by precisely replacing 6 Mb of mouse Ig heavy and kappa light variable region germ-line gene segments with their human counterparts while leaving the mouse constant regions intact, using a unique in situ humanization approach. We reasoned the introduced human variable region gene segments would function indistinguishably in their new genetic location, whereas the retained mouse constant regions would allow for optimal interactions and selection of the resulting antibodies within the mouse environment. We show that these mice, termed VelocImmune mice because they were generated using VelociGene technology, efficiently produce human: mouse hybrid antibodies (that are rapidly convertible to fully human antibodies) and have fully functional humoral immune systems indistinguishable from those of WT mice. The efficiency of the VelocImmune approach is confirmed by the rapid progression of 10 different fully human antibodies into human clinical trials.