Development of novel adoptive immunotherapy of malignant effusions using tumor cells genetically engineered to secrete interleukin-2

恶性积液是致死性癌症患者最常见的并发症之一，近年来建立有效的恶性积液治疗方法已成为世界各国关注的焦点。我们已经尝试开发一种新的恶性积液过继免疫疗法，利用肿瘤细胞刺激淋巴细胞分泌白细胞介素(IL)-2。首先，建立肿瘤细胞系（2个胃癌细胞系、2个结肠癌细胞系和1个胰腺癌细胞系），用人IL-2基因逆转录转导。mRNA分析检测转基因，ELISA和生物测定特异性IL-2检测IL-2分泌。其次，用mytomicin C灭活这些肿瘤细胞，并在体外刺激患者淋巴细胞。淋巴细胞进一步用固体相cd3 /IL-2系统培养，扩增1000倍。淋巴细胞对肿瘤细胞具有明显的细胞毒性。最后对3例恶性积液患者进行了安全的治疗，其中2例获得了临床疗效。提示过继免疫疗法使用肿瘤细胞基因工程刺激淋巴细胞分泌白细胞介素(IL) -2可能有效治疗恶性积液。需要进一步的经验来确定这种新型治疗方式的有效性。

Malignant effusion is one of the most frequent complications in patients with fatal cancer and the establishment of effective treatment for malignant effusions is recently focuced in the world. We have attempted to develop the novel adoptive immunotherapy of malignant effusions using lymphocytes stimulated with tumor cells genetically engineered to secrete interleukin (IL)-2.First, tumor cell lines (2 gastric, 2 colon and 1 pancreatic cancer cell lines) were established and retrovirally transduced with human IL-2 gene. Transgene was detected by mRNA analysis and IL-2 secretion was confirmed by ELISA and bioassay specific IL-2. Second, these tumor cells were inactivated with mytomicin C and subjected for in vitro stimulation of patients' lymphocytes. The lymphocytes were further cultured with solid phase-CD3/IL-2 system which could make 1,000-fold expansion. The lymphocytes had a significantly potent cytotoxicity againt tumor cells. Finally, 3 patients with malignant effusion were treated with this system in safe and clinical efficacy was obtained in 2 patients.It is suggested that adoptive immunotherapy using lymphocytes stimulated with tumor cells genetically engineered to secrete interleukin (IL) -2 may effective for treatment of malignant effusions. Further experiences are warranted to make sure the efficacy of this novel treatment modality.