Topalian S L, Muul L M, Solomon D, Rosenberg S A
J Immunol Methods. 1987 Aug 24;102(1):127-41. doi: 10.1016/s0022-1759(87)80018-2.
The potential utility of tumor-infiltrating lymphocytes (TIL) in the adoptive immunotherapy of human tumors has been suggested by murine experiments showing these cells to be 50-100 times more powerful than LAK cells in treating advanced metastatic disease. A method for the large-scale expansion of human TIL for the use of these cells in clinical trials is described in this report. TIL were successfully expanded on an experimental scale from 24 of 25 consecutive human tumors, including six melanomas, ten sarcomas, and eight adenocarcinomas. Tumors were digested enzymatically to yield single cell suspensions which were cultured in RPMI 1640 medium with 10% human serum and 1000 U/ml recombinant interleukin-2. Lymphocytes constituted from 3% to 74% of single cell tumor suspensions, and expanded from 2.9-fold to 9.1 X 10(8)-fold over a culture period ranging from 14 to 100 days. Nine of 24 TIL cultures lysed fresh autologous tumor targets in 4 h chromium release assays. Cell surface phenotyping identified cultured TIL as activated cytotoxic/suppressor T cells. Subsequently, large-scale expansion of TIL was successful in generating more than 10(10) lymphocytes in five of eight consecutive cases. Clinical trials employing the adoptive transfer of expanded TIL to patients with metastatic disease have begun.
小鼠实验表明肿瘤浸润淋巴细胞（TIL）在人类肿瘤的过继性免疫治疗中具有潜在效用，这些实验显示在治疗晚期转移性疾病时，这些细胞的效力比LAK细胞强50至100倍。本报告描述了一种大规模扩增人TIL以供这些细胞用于临床试验的方法。TIL在实验规模上成功地从25例连续的人类肿瘤中的24例扩增得到，包括6例黑色素瘤、10例肉瘤和8例腺癌。肿瘤经酶消化产生单细胞悬液，将其在含有10%人血清和1000 U/ml重组白细胞介素-2的RPMI 1640培养基中培养。淋巴细胞占单细胞肿瘤悬液的3%至74%，在14至100天的培养期内扩增了2.9倍至9.1×10⁸倍。在4小时铬释放试验中，24例TIL培养物中有9例裂解了新鲜的自体肿瘤靶标。细胞表面表型分析确定培养的TIL为活化的细胞毒性/抑制性T细胞。随后，在连续8例中的5例中成功进行了TIL的大规模扩增，产生了超过10¹⁰个淋巴细胞。采用将扩增的TIL过继转移给转移性疾病患者的临床试验已经开始。
