Generation of dual specific bivalent BiTEs (dbBIspecific T-cell engaging antibodies) for cellular immunotherapy
Generation of dual specific bivalent BiTEs (dbBIspecific T-cell engaging antibodies) for cellular immunotherapy
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DOI：
10.1186/s12885-019-6056-8
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发表时间：
2019-09-05
期刊：
影响因子：
3.8
通讯作者：
Shively, John E.
中科院分区：
文献类型：
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作者：
Kujawski, Maciej;Li, Lin;Shively, John E.
双特异性T细胞接合抗体（BiTES），包含双重抗-CD 3和抗-肿瘤抗原scFv片段，是用于治疗癌症的重要治疗剂。BiTES的双scFv构建体需要适当的蛋白质折叠，而它们的小分子尺寸导致快速的肾清除。方法将完整的（150 kDa）抗CEA的抗肿瘤抗原抗体以高产量（约150 kDa）连接到细胞上。30%）与完整的（150 kDa）抗鼠和抗人CD 3抗体连接，以形成双特异性二价BiTES（dbBiTES，300 kDa）。dbBiTE在体外通过EM、流式细胞术和细胞毒性进行测试，在体内通过PET肿瘤成像和重定向T细胞疗法进行测试。结果联锁铰链区是兼容的结构模型，适合300 kDa的颗粒的电子显微镜照片。与完整的抗CEA抗体相比，dbBiTES表现出高的体外细胞毒性、高的体内肿瘤靶向，如PET成像所证明的，以及重定向的dbBiTE包被的T细胞（1微克/1000万个细胞），其在CEA转基因小鼠中体内杀死CEA（+）靶细胞。结论dbBiTE介导的T细胞治疗是一种有效的靶向杀伤肿瘤细胞的方法。
Background Bispecific T-cell engaging antibodies (BiTES), comprising dual anti-CD3 and anti-tumor antigen scFv fragments, are important therapeutic agents for the treatment of cancer. The dual scFv construct for BiTES requires proper protein folding while their small molecular size leads to rapid kidney clearance. Methods An intact (150 kDa) anti-tumor antigen antibody to CEA was joined in high yield (ca. 30%) to intact (150 kDa) anti-murine and anti-human CD3 antibodies using hinge region specific Click chemistry to form dual-specific, bivalent BiTES (dbBiTES, 300 kDa). dbBiTEs were tested in vitro by EM, flow cytometry and cell cytoxicity and in vivo by PET tumor imaging and redirected T-cell therapy. Results The interlocked hinge regions are compatible with a structural model that fits the electron micrographs of 300 kDa particles. Compared to intact anti-CEA antibody, dbBiTES exhibit high in vitro cytotoxicity, high in vivo tumor targeting as demonstrated by PET imaging, and redirected dbBiTE coated T-cells (1 microgram/10 million cells) that kill CEA(+) target cells in vivo in CEA transgenic mice. Conclusion dbBiTE redirected T-cell therapy is a promising, efficient approach for targeting and killing cancer cells.