Transfer of a TCR gene derived from a patient with a marked antitumor response conveys highly active T-cell effector functions
Transfer of a TCR gene derived from a patient with a marked antitumor response conveys highly active T-cell effector functions
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DOI：
10.1089/hum.2005.16.457
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发表时间：
2005-04-01
影响因子：
4.2
通讯作者：
Morgan, RA
中科院分区：
文献类型：
--
作者：
Hughes, MS;Yu, YYL;Morgan, RA
高反应性抗MART-1 T细胞受体的α和β链的基因从T淋巴细胞中分离，所述T淋巴细胞介导转移性黑素瘤患者体内肿瘤消退。将这些基因克隆并插入基于MSCV的逆转录病毒载体中。转导后，在抗CD 3抗体刺激的原代T淋巴细胞中证实了大于50%的基因转移效率。TCR基因转导的T细胞的特异性和生物活性通过T细胞与用MART-1肽脉冲的刺激细胞共培养后的细胞因子产生来确定。干扰素-γ和粒细胞巨噬细胞集落刺激因子（GM-CSF）的产生与高活性MART-1特异性外周血淋巴细胞（PBL）在产生的细胞因子的量方面相当，并且转导的细胞在类似于细胞毒性T淋巴细胞（CTL）克隆的稀释度下识别肽脉冲细胞。通过脱粒标志物CD 107 a的动员、细胞溶解、细胞因子产生和HLA-A2阳性但非HLA-A2阴性黑色素瘤细胞系的增殖，证明了人类白细胞抗原（HLA）I类限制性识别。当用TCR基因转导肿瘤浸润淋巴细胞（TIL）时，获得了类似的数据，将以前的非反应性细胞转化为肿瘤反应性细胞。因此，TCR转导的T细胞是用于评估癌症患者的细胞转移疗法的有吸引力的候选者。
The genes for the alpha and beta chains of a highly reactive anti-MART-1 T-cell receptor were isolated from T-lymphocytes that mediated in vivo regression of tumor in a patient with metastatic melanoma. These genes were cloned and inserted into MSCV-based retroviral vectors. After transduction, greater than 50% gene transfer efficiency was demonstrated in primary T-lymphocytes stimulated by an anti-CD3 antibody. The specificity and biologic activity of TCR gene-transduced T-cells was determined by cytokine production after coculture of T-cells with stimulator cells pulsed with MART-1 peptide. The production of interferon-gamma and granulocyte macrophage-colony stimulating factor (GM-CSF) was comparable to highly active MART-1 specific peripheral blood lymphocytes (PBL) in the amount of cytokine produced and transduced cells recognized peptide pulsed cells at dilutions similar to cytotoxic T lymphocyte (CTL) clones. Human leukocyte antigen (HLA) class I restricted recognition was demonstrated by mobilization of degranulation marker CD107a, by cell lysis, by cytokine production, and by proliferation in the presence of HLA-A2-positive but not HLA-A2-negative melanoma cell lines. Similar data was obtained when tumor-infiltrating lymphocytes (TIL) were transduced with the TCR genes, converting previously nonreactive cells to tumor reactive cells. TCR-transduced T-cells are thus attractive candidates for evaluation in cell transfer therapies of patients with cancer.