Stolfi V M, Milsom J W, Finke J H, Fazio V W, Fiocchi C
Department of Colorectal Surgery, Cleveland Clinic Foundation, Ohio 44195.
J Surg Res. 1992 Jan;52(1):39-45. doi: 10.1016/0022-4804(92)90276-6.
Adoptive immunotherapy with tumor-infiltrating lymphocytes (TIL) and interleukin-2 (IL2) can induce regression of tumor metastases in animal models and in human metastatic malignant melanoma. We investigated the potential of colorectal cancer TIL as a source of killer cells and the effect of tumor necrosis factor alpha (TNF alpha) in combination with IL2 on their cytotoxic activity. Tumor-infiltrating lymphocytes were isolated from surgical specimens using a mechanical and enzymatic dissociation process. Autologous lamina propria mononuclear cells (LPMC) were used as control. Tumor-infiltrating lymphocytes and LPMC were cultured in the presence of IL2 with/without TNF alpha (1000 U/ml each) for 5 to 8 weeks. Cytotoxicity (% lysis) was tested against Daudi target cells in a 4-hr 51Cr-release assay. The combination of IL2 and TNF alpha resulted in a significantly greater-fold expansion of TIL than IL2 alone (P less than 0.01). Lamina propria mononuclear cells expanded less than TIL, and TNF alpha had an inhibitory effect on their growth (P less than 0.05). Tumor-infiltrating lymphocytes and LPMC showed comparable cytotoxicity when cultured with IL2 alone. However, the addition of TNF alpha augmented the killer activity of TIL while inhibiting that of LPMC (P = 0.035). These results indicate that TNF alpha selectively increases the IL2-induced growth and cytotoxic function of colorectal cancer TIL, but not those of gut mucosal lymphoid cells, suggesting that TIL and LMPC differ in their response to TNF alpha. Therefore, this combination of cytokines may hold more promise than single agents for the immunotherapy of colorectal cancers with TIL.
采用肿瘤浸润淋巴细胞（TIL）和白细胞介素-2（IL2）进行过继性免疫疗法可在动物模型和人类转移性恶性黑色素瘤中诱导肿瘤转移灶消退。我们研究了结直肠癌TIL作为杀伤细胞来源的潜力，以及肿瘤坏死因子α（TNFα）与IL2联合使用对其细胞毒性活性的影响。使用机械和酶解过程从手术标本中分离肿瘤浸润淋巴细胞。自体固有层单核细胞（LPMC）用作对照。肿瘤浸润淋巴细胞和LPMC在有/无TNFα（各1000 U/ml）存在的情况下于IL2中培养5至8周。在4小时的51Cr释放试验中针对Daudi靶细胞测试细胞毒性（%裂解率）。IL2和TNFα联合使用导致TIL的扩增倍数显著高于单独使用IL2（P<0.01）。固有层单核细胞的扩增少于TIL，且TNFα对其生长有抑制作用（P<0.05）。单独用IL2培养时，肿瘤浸润淋巴细胞和LPMC表现出相当的细胞毒性。然而，添加TNFα增强了TIL的杀伤活性，同时抑制了LPMC的杀伤活性（P = 0.035）。这些结果表明，TNFα选择性地增加IL2诱导的结直肠癌TIL的生长和细胞毒性功能，但不增加肠道黏膜淋巴细胞的生长和细胞毒性功能，提示TIL和LMPC对TNFα的反应不同。因此，对于用TIL进行结直肠癌免疫治疗，这种细胞因子组合可能比单一药物更有前景。
