Generation of Human Antigen-Specific Monoclonal IgM Antibodies Using Vaccinated "Human Immune System'' Mice
Generation of Human Antigen-Specific Monoclonal IgM Antibodies Using Vaccinated "Human Immune System'' Mice
复制标题
DOI：
10.1371/journal.pone.0013137
复制
发表时间：
2010-10-04
期刊：
影响因子：
3.7
通讯作者：
Spits, Hergen
中科院分区：
文献类型：
--
作者：
Becker, Pablo D.;Legrand, Nicolas;Spits, Hergen
背景资料：抗体的被动转移不仅提供针对疾病的即时短期保护，而且还可以用作治疗工具。然而，鼠单克隆抗体（mAb）的“人源化”是耗时且昂贵的过程，其具有潜在改变抗原特异性和/或亲和力的固有缺点。人B细胞的永生化代表了获得人mAb的替代方案，但依赖于来自接种疫苗的个体或康复期患者的生物样品的可用性。在这项工作中，我们描述了一种新的方法来产生完全人单克隆抗体相结合的人源化小鼠模型与新的B细胞immortalization technique.Methodology/Principal Findings：移植后与CD 34（+）CD 38（-）人造血祖细胞，BALB/c Rag 2（-/-）IL-2 R γ c（-/-）小鼠获得了人类免疫系统和港口B细胞与不同的IgM库。然后用两种商业疫苗抗原，破伤风类毒素和B型肝炎表面抗原免疫“人免疫系统”小鼠。用人B细胞淋巴瘤（BCL）-6和BCL-XL基因逆转录病毒转导分选的人CD 19（+）CD 27（+）B细胞，随后在CD 40-配体和IL-21存在下培养。该方法允许产生分泌免疫球蛋白的稳定的B细胞受体阳性B细胞。我们恢复稳定的B细胞克隆，产生特异性的破伤风类毒素和肝炎B表面抗原的IgM抗体，respectively.Conclusion/Significance：这项工作提供了这种新的方法的实用性的概念验证的基础上的人源化小鼠的免疫的快速生成的人单克隆抗体对广泛的抗原。
Background: Passive transfer of antibodies not only provides immediate short-term protection against disease, but also can be exploited as a therapeutic tool. However, the 'humanization' of murine monoclonal antibodies (mAbs) is a time-consuming and expensive process that has the inherent drawback of potentially altering antigenic specificity and/or affinity. The immortalization of human B cells represents an alternative for obtaining human mAbs, but relies on the availability of biological samples from vaccinated individuals or convalescent patients. In this work we describe a novel approach to generate fully human mAbs by combining a humanized mouse model with a new B cell immortalization technique.Methodology/Principal Findings: After transplantation with CD34(+)CD38(-) human hematopoietic progenitor cells, BALB/c Rag2(-/-)IL-2R gamma c(-/-) mice acquire a human immune system and harbor B cells with a diverse IgM repertoire. "Human Immune System'' mice were then immunized with two commercial vaccine antigens, tetanus toxoid and hepatitis B surface antigen. Sorted human CD19(+)CD27(+) B cells were retrovirally transduced with the human B cell lymphoma (BCL)-6 and BCL-XL genes, and subsequently cultured in the presence of CD40-ligand and IL-21. This procedure allows generating stable B cell receptor-positive B cells that secrete immunoglobulins. We recovered stable B cell clones that produced IgM specific for tetanus toxoid and the hepatitis B surface antigen, respectively.Conclusion/Significance: This work provides the proof-of-concept for the usefulness of this novel method based on the immunization of humanized mice for the rapid generation of human mAbs against a wide range of antigens.