Elia Angela R, Circosta Paola, Sangiolo Dario, Bonini Chiara, Gammaitoni Loretta, Mastaglio Sara, Genovese Pietro, Geuna Massimo, Avolio Fabio, Inghirami Giorgio, Tarella Corrado, Cignetti Alessandro
1 Molecular Biotechnology Center, University of Torino , 10126 Turin, Italy .
Hum Gene Ther. 2015 Apr;26(4):220-31. doi: 10.1089/hum.2014.112. Epub 2015 Apr 13.
Cytokine-induced killer (CIK) cells consist of a heterogeneous population of polyclonal T lymphocytes displaying NK phenotype and HLA-unrestricted cytotoxic activity against a broad range of tumors. We sought to determine whether transduction of CIK cells with T cell receptor (TCR) genes specific for tumor-associated antigens could generate effector cells endowed with a double mechanism of tumor recognition. HLA-A2-restricted TCR-transduced (TD) CIK directed against the melanoma antigens Mart1 and NY-ESO1 were generated by lentiviral transduction and successfully expanded over a 3-4-week period. TD-CIK cells were both CD3(+)/CD56(-) and CD3(+)/CD56(+) (31±8% and 59±9%, respectively), indicating that both major histocompatibility complex (MHC)-restricted T cells and MHC-unrestricted CIK could be targeted by lentiviral transduction. At the end of the culture, the majority of both unmodified and TD-CIK displayed an effector memory phenotype, without considerable expression of replicative senescence and exhaustion markers. Functionally, TD-CIK specifically recognized tumor cells expressing the relevant antigen as well as maintained their MHC-unrestricted tumor activity. The cytotoxic activity of TD-CIK against HLA-A2(+) melanoma cell lines was significantly higher than the untransduced counterparts at a low effector:target ratio (cytotoxic activity of TD-CIK was from 1.9- to 4.3-fold higher than untransduced counterparts). TD-CIK were highly proficient in releasing high amount of IFN-γ upon antigen-specific stimulation and were able to recognize primary melanoma targets. In conclusion, we showed that (1) the reproducibility and simplicity of CIK transduction and expansion might solve the problem of obtaining adequate numbers of potent antitumor effector cells for adoptive immunotherapy; (2) the presence of both terminal effectors as well as of less differentiated progenitors might confer them long survival in vivo; and (3) the addition of an MHC-restricted antigen recognition allows not only targeting tumor surface antigens but also a wider range of cytoplasmic or nuclear antigens, involved in tumor proliferation and survival. TD-CIK cells with a double mechanism of tumor recognition are an attractive and alternative tool for the development of efficient cell therapeutic strategies.
细胞因子诱导的杀伤（CIK）细胞由一群异质性的多克隆T淋巴细胞组成，这些细胞表现出自然杀伤（NK）细胞表型，并对多种肿瘤具有HLA非限制性细胞毒性活性。我们试图确定用针对肿瘤相关抗原的T细胞受体（TCR）基因转导CIK细胞是否能产生具有双重肿瘤识别机制的效应细胞。通过慢病毒转导产生了针对黑色素瘤抗原Mart1和NY-ESO1的HLA-A2限制性TCR转导（TD）CIK细胞，并在3至4周的时间内成功扩增。TD-CIK细胞既有CD3(+)/CD56(-)细胞，也有CD3(+)/CD56(+)细胞（分别为31±8%和59±9%），这表明主要组织相容性复合体（MHC）限制性T细胞和MHC非限制性CIK细胞都可以被慢病毒转导。培养结束时，未修饰的CIK细胞和TD-CIK细胞大多表现出效应记忆表型，没有明显表达复制性衰老和耗竭标志物。在功能上，TD-CIK细胞能特异性识别表达相关抗原的肿瘤细胞，并保持其MHC非限制性肿瘤活性。在低效应细胞与靶细胞比例下，TD-CIK细胞对HLA-A2(+)黑色素瘤细胞系的细胞毒性活性显著高于未转导的对照细胞（TD-CIK细胞的细胞毒性活性比未转导的对照细胞高1.9至4.3倍）。TD-CIK细胞在抗原特异性刺激后能高效释放大量干扰素-γ，并能够识别原发性黑色素瘤靶细胞。总之，我们表明：（1）CIK细胞转导和扩增的可重复性和简便性可能解决过继性免疫治疗中获得足够数量强效抗肿瘤效应细胞的问题；（２）终末效应细胞以及分化程度较低的祖细胞的存在可能赋予它们在体内较长的存活时间；（３）添加MHC限制性抗原识别不仅可以靶向肿瘤表面抗原，还可以靶向更广泛的参与肿瘤增殖和存活过程的细胞质或核抗原。具有双重肿瘤识别机制的TD-CIK细胞是开发高效细胞治疗策略的一种有吸引力的替代工具。
