Haas G P, Solomon D, Rosenberg S A
Surgery Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892.
Cancer Immunol Immunother. 1990;30(6):342-50. doi: 10.1007/BF01786883.
Tumor-infiltrating lymphocytes (TIL) were isolated from 15 of 20 surgical specimens of transitional cell carcinoma of the urinary bladder, prostate cancer, testicular cancer, Wilms tumor and adrenal cancer. Expansion was carried out in four different culture conditions, each containing 1000 U/ml interleukin-2: RPMI medium with or without 20% (by volume) of lymphokine-activated killer cell (LAK) supernatant and AIM V medium with or without 20% LAK supernatant. The resultant cell populations were then assayed for cytotoxicity against a variety of autologous and allogeneic tumor targets and phenotypic analysis was performed with fluorescein-labeled monoclonal antibodies. TIL growth was unrelated to the initial percentage of lymphocytes or tumor cells present in the enzymatically dispersed specimens or whether fresh or cryopreserved tissue was utilized. Better growth was seen in AIM V than in RPMI medium (P = 0.013); the beneficial effect of the addition of LAK supernatant to RPMI was indicated (P = 0.065), and the addition of LAK supernatant to AIM V did not improve the ability to culture TIL (P = 0.5) from these cancers. TIL in long-term culture were predominantly CD3+. The ratio of CD4+/CD8+ cells varied with time in culture and culture medium, but most cultures eventually became CD4+. Cells bearing B cell, natural killer cell, and macrophage markers disappeared early in culture. Overall 14/15 TIL samples were lytic against one of the autologous and allogeneic targets tested, but specific lysis against the autologous tumor from which it was derived was seen in only one TIL culture originating from a bladder cancer. Our results suggest that TIL can be expanded to therapeutic levels from a variety of urological malignancies and that their potential role in future therapy should be further explored.
从20份手术标本(包括膀胱癌、前列腺癌、睾丸癌、肾母细胞瘤和肾上腺癌的移行细胞癌)中选取15份分离肿瘤浸润淋巴细胞(TIL)。在四种不同的培养条件下进行扩增,每种条件均含有1000 U/ml白细胞介素-2:含或不含20%(体积分数)淋巴因子激活的杀伤细胞(LAK)上清液的RPMI培养基,以及含或不含20% LAK上清液的AIM V培养基。然后检测所得细胞群体对多种自体和异体肿瘤靶标的细胞毒性,并用荧光素标记的单克隆抗体进行表型分析。TIL的生长与酶分散标本中淋巴细胞或肿瘤细胞的初始百分比无关,也与使用新鲜组织还是冷冻保存组织无关。在AIM V培养基中观察到的生长情况优于RPMI培养基(P = 0.013);向RPMI培养基中添加LAK上清液显示出有益效果(P = 0.065),而向AIM V培养基中添加LAK上清液并未提高培养这些癌症来源的TIL的能力(P = 0.5)。长期培养的TIL主要为CD3+。CD4+/CD8+细胞的比例随培养时间和培养基而变化,但大多数培养物最终变为CD4+。带有B细胞、自然杀伤细胞和巨噬细胞标志物的细胞在培养早期消失。总体而言,14/15的TIL样本对所测试的一种自体和异体靶标具有细胞毒性,但仅在源自膀胱癌的一个TIL培养物中观察到对其来源的自体肿瘤的特异性细胞毒性。我们的结果表明,TIL可从多种泌尿生殖系统恶性肿瘤中扩增至治疗水平,其在未来治疗中的潜在作用应进一步探索。