Lafreniere R, Borkenhagen K, Bryant L D, Ng E
Oncology Research Group, University of Calgary, Alberta, Canada.
J Biol Response Mod. 1989 Jun;8(3):238-51.
Tumor-infiltrating lymphocytes (TILs) have potent antitumor effects in murine models of advanced disease. Although these cells are 50-100 times more potent than lymphokine-activated killer cells against micrometastases, their antitumor benefits are virtually nonexistent against large tumor burdens unless cyclophosphamide is added to the immunotherapy regimen. In an effort to determine the effects of cyclophosphamide on TILs obtained from tumor-bearing animals, we harvested tumors from animals having received either 0, 50, or 100 mg/kg cyclophosphamide intravenously and investigated the expansion, cytotoxicity, and phenotypic expression of these TILs co-cultured in vitro with recombinant interleukin-2. TILs obtained from animals given cyclophosphamide, demonstrated a greater fold expansion than TILs obtained from normal animals (mean fold expansion on day 59 of culture: 85, 400, and 150 for TILs obtained from animals given 0, 50, and 100 mg/kg cyclophosphamide, n = 3 consecutive experiments). In addition, these TILs demonstrated enhanced cytotoxicity compared to controls [effector to target ratio 4:1, day 16 TILs, % lysis: 24, 35, and 45%, cyclophosphamide 0, 50, and 100 mg/kg, respectively, against the MCA-102 sarcoma, natural killer (NK) insensitive tumor; 29, 38, and 56% against the YAC-1 lymphoma, NK sensitive tumor]. Similar results were seen with day 34 and day 59 TILs. When phenotypic analysis was performed, TILs obtained from animals given cyclophosphamide consistently demonstrated a greater percent expression of the Thy1.2 and Lyt-2 antigens up to day 59 of culture when the experiments were terminated. The NK cell marker 49H.8 was expressed on the majority of TILs and its expression did not change with respect to the cyclophosphamide concentration. The increase in TIL number and cytotoxicity seen with cyclophosphamide given intravenously before tumor harvest could have important ramifications for human immunotherapy with TILs.
肿瘤浸润淋巴细胞（TILs）在晚期疾病的小鼠模型中具有强大的抗肿瘤作用。尽管这些细胞针对微转移灶的效力比淋巴因子激活的杀伤细胞强50至100倍，但除非在免疫治疗方案中加入环磷酰胺，否则它们对大肿瘤负荷几乎没有抗肿瘤益处。为了确定环磷酰胺对从荷瘤动物获得的TILs的影响，我们从静脉注射了0、50或100mg/kg环磷酰胺的动物身上采集肿瘤，并研究了这些TILs与重组白细胞介素-2在体外共培养时的扩增、细胞毒性和表型表达。从给予环磷酰胺的动物获得的TILs，其扩增倍数比从正常动物获得的TILs更大（培养第59天的平均扩增倍数：给予0、50和100mg/kg环磷酰胺的动物所获得的TILs分别为85、400和150，n = 3次连续实验）。此外，与对照组相比，这些TILs表现出增强的细胞毒性[效靶比4:1，第16天的TILs，裂解率%：分别为24、35和45%，环磷酰胺剂量为0、50和100mg/kg，针对MCA - 102肉瘤（自然杀伤（NK）不敏感肿瘤）；针对YAC - 1淋巴瘤（NK敏感肿瘤）分别为29、38和56%]。在第34天和第59天的TILs中也观察到了类似结果。当进行表型分析时，在实验终止时，直到培养第59天，从给予环磷酰胺的动物获得的TILs始终显示出更高百分比的Thy1.2和Lyt - 2抗原表达。NK细胞标志物49H.8在大多数TILs上表达，其表达相对于环磷酰胺浓度没有变化。在肿瘤收获前静脉给予环磷酰胺后，TIL数量和细胞毒性的增加可能对人类TIL免疫治疗具有重要意义。
