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Updated: Jun 30, 2025

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在体内亲和力成熟的小鼠B细胞被重新编程以表达人类抗体

Yiming Yin^1,2, Yan Guo³, Yuxuan Jiang⁴

¹Department of Pediatrics, Boston Children's Hospital, Harvard Medical School, Boston, MA, USA. yiming.yin@childrens.harvard.edu.

Nature biomedical engineering

|March 15, 2024

概括

这项研究介绍了一种新的CRISPR编辑方法,用于小鼠B细胞表达人类抗体,保持关键功能,如亲和力成熟. 这一进步有助于开发疫苗和抗体疗法,治疗包括艾滋病毒和SARS-CoV-2在内的疾病.

科学领域:

免疫学免疫学免疫学
分子生物学分子生物学
生物技术是生物技术.

背景情况:

目前的CRISPR编辑小鼠B细胞用于人类抗体表达,破坏了重链局部,损害了体质突变和亲和力成熟.
开发保存B细胞功能的方法对于评估疫苗和创造有效的抗体疗法至关重要.

研究的目的:

开发一种基于CRISPR的新策略,同时用B细胞中的人类抗体变量链替换小鼠重链和卡帕链.
为了在基因编辑后保持基本的B细胞功能,包括体质突变和亲和力成熟.
评估这种方法在产生针对HIV-1和SARS-CoV-2的强效中和抗体方面的有效性.

主要方法:

利用Cas12a介导的基因编辑,在每个位点进行单一切割,5'同质臂与远端V段相辅相成.
编辑小鼠B细胞以表达人类广泛中和抗体 (HIV-1 10-1074和VRC26.25-y).
通过收养将编辑的B细胞转移到小鼠体内,接种疫苗,并分析抗体反应.

主要成果:

这种新的编辑策略成功地保留了B细胞功能,使得强大的超突变和强大的中和性血生成成为可能.
编辑的表达HIV-1抗体10-1074的B细胞产生了增强对各种HIV-1菌株中和性的变体,保持了低多活性和长半衰期.
这种方法改善了抗SARS-CoV-2抗体对抗Omicron变异的效力.

结论:

内源性重链和卡帕链的直接,同时更换可保持B细胞功能,促进体内亲和力成熟.
这种方法可以开发广泛中和,强效和生物可用的人类抗体,用于治疗.
该战略有望推动疫苗开发和针对传染病的基于抗体的疗法.

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