Krishnadas Deepa Kolaseri, Wang Yali, Sundaram Kumaran, Bai Fanqi, Lucas Kenneth G
Department of Pediatrics, Division of Hematology/Oncology, University of Louisville, Louisville, KY, USA.
Tumour Biol. 2017 Jul;39(7):1010428317701309. doi: 10.1177/1010428317701309.
The cancer germline antigens MAGE-A1, MAGE-A3, and NY-ESO-1 can be used to target relapsed or therapy-resistant malignant solid tumors, and previous studies have demonstrated that these antigens can be epigenetically upregulated on the surface of tumor cells following exposure to low-dose demethylating chemotherapy agents, such as decitabine. The extent to which cancer germline antigen cytotoxic T lymphocytes can be reliably expanded from healthy donors has not been well characterized, specifically in terms of whether these T cells consistently kill antigen-bearing targets or simply produce interferon-γ in the presence of the antigen. Cancer germline antigen cytotoxic T lymphocytes were generated using conventional method and high-density lymphocyte culture method. We demonstrate that there is no difference in the extent of antigen-specific killing with or without CD25 depletion when interleukin-21 is added to the cultures. Cancer germline antigen-specific killer cells could be expanded from 8/12 healthy donors using overlapping peptide mixes derived from MAGE-A1, MAGE-A3, and NY-ESO-1 and from 7/9 healthy donors using HLA-restricted epitopes. Furthermore, cytotoxic T lymphocyte derived from 4/5 patients displayed specific cytotoxicity of target cells expressing respective cancer germline antigen and HLA partially matched tumor lines. High-density lymphocyte culture prior to stimulation with cancer germline antigen peptides resulted in antigen-specific cytotoxic T lymphocyte from healthy donors and patients from whom cancer germline antigen cytotoxic T lymphocyte culture with conventional methods was not feasible. These data demonstrate that MAGE-A1-, MAGE-A3-, and NY-ESO-1-specific T cells with antigen-specific cytotoxicity can be cultured from healthy donors and patient-derived cells making adoptive immunotherapy with these cytotoxic T lymphocyte feasible.
癌症种系抗原MAGE-A1、MAGE-A3和NY-ESO-1可用于靶向复发或耐药的恶性实体瘤，先前的研究表明，在暴露于低剂量去甲基化化疗药物（如地西他滨）后，这些抗原可在肿瘤细胞表面发生表观遗传上调。从健康供体中可靠扩增癌症种系抗原细胞毒性T淋巴细胞的程度尚未得到充分表征，特别是这些T细胞是持续杀伤携带抗原的靶细胞，还是仅在抗原存在时产生干扰素-γ。使用传统方法和高密度淋巴细胞培养方法产生癌症种系抗原细胞毒性T淋巴细胞。我们证明，当向培养物中添加白细胞介素-21时，有无CD25去除的抗原特异性杀伤程度没有差异。使用源自MAGE-A1、MAGE-A3和NY-ESO-1的重叠肽混合物，可从8/12名健康供体中扩增出癌症种系抗原特异性杀伤细胞，使用HLA限制性表位可从7/9名健康供体中扩增出该细胞。此外，源自4/5名患者的细胞毒性T淋巴细胞对表达相应癌症种系抗原和HLA部分匹配肿瘤系的靶细胞显示出特异性细胞毒性。在用癌症种系抗原肽刺激之前进行高密度淋巴细胞培养，可从健康供体和患者来源的细胞中产生抗原特异性细胞毒性T淋巴细胞，而用传统方法进行癌症种系抗原细胞毒性T淋巴细胞培养对这些患者不可行。这些数据表明，具有抗原特异性细胞毒性的MAGE-A1、MAGE-A3和NY-ESO-1特异性T细胞可从健康供体和患者来源的细胞中培养出来，使得用这些细胞毒性T淋巴细胞进行过继性免疫治疗成为可能。
