Hammel J M, Tuck M K, Hain J M, Chang A E, Sondak V K
Department of Surgery, University of Michigan Medical Center, Ann Arbor.
J Immunother Emphasis Tumor Immunol. 1994 Jul;16(1):1-12. doi: 10.1097/00002371-199407000-00001.
Adoptive immunotherapy using in vitro activated T-cells can mediate the destruction of metastatic tumor deposits in animal models. These antitumor effector cells can be generated from mice with subcutaneous tumor by the sequential in vitro activation of tumor-draining lymph node (TDLN) cells with monoclonal antibody to the T-cell receptor complex (anti-CD3) and expansion in low concentrations of interleukin-2 (IL-2). In this animal model, the concomitant presence of visceral tumor can suppress the sensitization of tumor-reactive TDLN cells. We investigated whether IL-1 alpha added during in vitro activation and/or expansion of TDLNs could augment their antitumor activity in adoptive therapy. Mice were inoculated subcutaneously with MCA 205 tumor. TDLN cells were harvested and activated in vitro with 1 microgram/ml anti-CD3 for 2 days (anti-CD3 phase), followed by expansion in 10 U/ml IL-2 for 3 days (IL-2 phase). Experimental cultures had IL-1 (10-10,000 U/ml) added in either or both phases. After the 5-day culture period, cells were counted to determine in vitro cellular proliferation and then adoptively transferred to mice bearing 3-day established lung metastases to assess in vivo antitumor efficacy. IL-1 added during the anti-CD3 or IL-2 phase did not alter in vitro cellular proliferation. The presence of IL-1 during the anti-CD3 phase led to the generation of cells that were significantly more therapeutically efficacious than cells generated in the absence of IL-1. The effect of IL-1 during anti-CD3 activation appeared to be dose dependent in the concentration range 10-1,000 U/ml. The addition of IL-1 during the IL-2 phase only did not enhance the antitumor reactivity of the activated cells. The beneficial effect of IL-1 during the anti-CD3 activation phase was specific for the tumor against which the TDLN had been initially sensitized; also, there was no evidence that non-tumor bearer lymphocytes developed significant antitumor reactivity when "activated" with IL-1 and anti-CD3. Furthermore, addition of IL-1 during the anti-CD3 activation phase abrogated suppression induced by the concomitant presence of lung metastases during subcutaneous tumor growth. IL-1 appears to up-regulate the in vitro activation of tumor-reactive lymphocytes derived from TDLN, in both the presence and the absence of visceral metastases.
利用体外活化的T细胞进行过继性免疫治疗可介导动物模型中转移性肿瘤沉积物的破坏。这些抗肿瘤效应细胞可从皮下接种肿瘤的小鼠中产生，方法是先用抗T细胞受体复合物的单克隆抗体（抗CD3）对肿瘤引流淋巴结（TDLN）细胞进行体外顺序活化，然后在低浓度白细胞介素-2（IL-2）中进行扩增。在这个动物模型中，内脏肿瘤的同时存在可抑制肿瘤反应性TDLN细胞的致敏。我们研究了在TDLN的体外活化和/或扩增过程中添加IL-1α是否能增强其在过继性治疗中的抗肿瘤活性。给小鼠皮下接种MCA 205肿瘤。收集TDLN细胞，用1微克/毫升抗CD3在体外活化2天（抗CD3阶段），随后在10单位/毫升IL-2中扩增3天（IL-2阶段）。实验培养物在一个或两个阶段添加IL-1（10 - 10,000单位/毫升）。在5天培养期后，计数细胞以确定体外细胞增殖，然后将其过继转移到已形成3天肺转移的小鼠中，以评估体内抗肿瘤疗效。在抗CD3或IL-2阶段添加IL-1均未改变体外细胞增殖。在抗CD3阶段存在IL-1导致产生的细胞在治疗上比在无IL-1情况下产生的细胞更有效。在抗CD3活化过程中，IL-1在10 - 1,000单位/毫升浓度范围内的作用似乎呈剂量依赖性。仅在IL-阶段添加IL-1并不能增强活化细胞的抗肿瘤反应性。在抗CD3活化阶段，IL-1的有益作用对TDLN最初致敏的肿瘤具有特异性；此外，没有证据表明非荷瘤淋巴细胞在用IL-1和抗CD3“活化”时会产生显著的抗肿瘤反应性。此外，在抗CD3活化阶段添加IL-1可消除皮下肿瘤生长过程中肺转移同时存在所诱导的抑制作用。无论是否存在内脏转移，IL-1似乎都能上调源自TDLN的肿瘤反应性淋巴细胞的体外活化。
