Lymphocyte Display: A Novel Antibody Selection Platform Based on T Cell Activation
Lymphocyte Display: A Novel Antibody Selection Platform Based on T Cell Activation
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DOI：
10.1371/journal.pone.0007174
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发表时间：
2009-09-24
期刊：
影响因子：
3.7
通讯作者：
Alvarez-Vallina, Luis
中科院分区：
文献类型：
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作者：
Alonso-Camino, Vanesa;Sanchez-Martin, David;Alvarez-Vallina, Luis
自其出现以来，展示技术已被证明可用于选择针对多种靶标的抗体;然而，用目前可用的平台选择的大多数抗体需要进一步修饰以用于人类，并且限于可获得的抗原。此外，这些平台不太适合于体内选择。我们在这里提出了一种新的基于细胞的抗体展示平台，它利用T淋巴细胞的功能能力。作为嵌合免疫受体（CIR）介导信号传导的一部分，T淋巴细胞表面上抗体的展示可以理想地将抗原-抗体相互作用与T细胞表型的可证实的变化联系起来，这是由于早期T细胞活化标志物CD 69的随后表达。在该概念验证中，使用经慢病毒转导以在表面上表达肿瘤特异性CIR的人T细胞系针对表达癌胚抗原的人肿瘤细胞系进行体外选择。基于CIR和肿瘤抗原之间的有效相互作用，我们证明了将CIR介导的活化与CD 69（+）T细胞的FACS分选相结合，可以分离肿瘤特异性细胞表面抗原的结合剂，两轮后富集因子至少为10（3）倍，从而产生表达肿瘤特异性CIR的同质T细胞群。
Since their onset, display technologies have proven useful for the selection of antibodies against a variety of targets; however, most of the antibodies selected with the currently available platforms need to be further modified for their use in humans, and are restricted to accessible antigens. Furthermore, these platforms are not well suited for in vivo selections. We present here a novel cell based antibody display platform, which takes advantage of the functional capabilities of T lymphocytes. The display of antibodies on the surface of T lymphocytes, as a part of a chimeric-immune receptor (CIR) mediating signaling, may ideally link the antigen-antibody interaction to a demonstrable change in T cell phenotype, due to subsequent expression of the early T cell activation marker CD69. In this proof-of-concept, an in vitro selection was carried out using a human T cell line lentiviral-transduced to express a tumor-specific CIR on the surface, against a human tumor cell line expressing the carcinoembryonic antigen. Based on an effective interaction between the CIR and the tumor antigen, we demonstrated that combining CIR-mediated activation with FACS sorting of CD69(+) T cells, it is possible to isolate binders to tumor specific cell surface antigen, with an enrichment factor of at least 10(3)-fold after two rounds, resulting in a homogeneous population of T cells expressing tumor-specific CIRs.