从表达组成性激活 STAT5 的 B 细胞中分离出的抗原特异性单克隆抗体。

Antigen-specific monoclonal antibodies isolated from B cells expressing constitutively active STAT5.

机构信息

Department of Cell Biology and Histology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands.

出版信息

PLoS One. 2011 Apr 15;6(4):e17189. doi: 10.1371/journal.pone.0017189.

BACKGROUND

Fully human monoclonal antibodies directed against specific pathogens have a high therapeutic potential, but are difficult to generate.

METHODOLOGY/PRINCIPAL FINDINGS: Memory B cells were immortalized by expressing an inducible active mutant of the transcription factor Signal Transducer and Activator of Transcription 5 (STAT5). Active STAT5 inhibits the differentiation of B cells while increasing their replicative life span. We obtained cloned B cell lines, which produced antibodies in the presence of interleukin 21 after turning off STAT5. We used this method to obtain monoclonal antibodies against the model antigen tetanus toxin.

CONCLUSIONS/SIGNIFICANCE: Here we describe a novel and relatively simple method of immortalizing antigen-specific human B cells for isolation of human monoclonal antibodies. These results show that STAT5 overexpression can be employed to isolate antigen specific antibodies from human memory B cells.

背景

针对特定病原体的全人源单克隆抗体具有很高的治疗潜力，但难以产生。

方法/主要发现：通过表达转录因子信号转导和转录激活因子 5（STAT5）的诱导型活性突变体，使记忆 B 细胞永生化。活性 STAT5 抑制 B 细胞的分化，同时增加其复制寿命。我们获得了克隆的 B 细胞系，在关闭 STAT5 后，在白细胞介素 21 的存在下产生针对模型抗原破伤风毒素的抗体。我们使用这种方法获得了针对破伤风毒素的单克隆抗体。

结论/意义：在这里，我们描述了一种新颖且相对简单的永生化抗原特异性人 B 细胞的方法，用于分离人单克隆抗体。这些结果表明，STAT5 的过表达可用于从人记忆 B 细胞中分离抗原特异性抗体。