Veelken H, Mackensen A, Lahn M, Köhler G, Becker D, Franke B, Brennscheidt U, Kulmburg P, Rosenthal F M, Keller H, Hasse J, Schultze-Seemann W, Farthmann E H, Mertelsmann R, Lindemann A
Department of Hematology/Oncology, Freiburg University Medical Center, Germany.
Int J Cancer. 1997 Jan 27;70(3):269-77. doi: 10.1002/(sici)1097-0215(19970127)70:3<269::aid-ijc4>3.0.co;2-v.
Tumor cells transfected to express immunostimulatory cytokines, or admixed with similarly modified bystander cells, are able to induce immune responses against unmodified tumor cells in animal models. For treatment of human patients, a vaccine composed of autologous tumor cells and IL-2-secreting allogeneic fibroblasts was developed. Autologous tumor cells were isolated from biopsy specimens. A clone (KMST 6.14) of an immortalized human fibroblast line that stably secreted 5290 IU IL-2 per 10(6) cells and per 24 hr was obtained by cationic lipofection with an expression construct for human IL-2 and Neo(r). Fifteen patients with refractory malignant tumors received 3-4 injections of irradiated KMST6.14 and autologous tumor cells in a phase-I clinical trial. Increasing transient inflammatory responses without systemic toxicity developed at vaccination sites and after injections with irradiated tumor cells only (p < 0.05). These sites contained a dense infiltrate of CD3+ T cells with numbers of CD4+ helper cells exceeding those of CD8+ cytotoxic T cells (CTL). CD8+ T-cell lines isolated from vaccination sites of 2 malignant melanoma patients but not of renal-cell carcinoma patients exhibited a dominant lytic activity against autologous tumor cells in vitro. CD8+ T-cell clones established from the vaccination site of 1 of 2 renal-cell carcinoma patients preferentially lysed autologous and partially matched allogeneic renal-cell carcinoma cells. In conclusion, a vaccine composed of IL-2 gene-transfected allogeneic fibroblasts and autologous tumor cells is able to enhance specific anti-tumor T-cell responses in vivo without major side-effects. Malignant melanoma and renal-cell carcinoma appear to be promising entities for testing of similar approaches in future therapeutic trials.
转染以表达免疫刺激细胞因子的肿瘤细胞，或与经类似修饰的旁观者细胞混合，在动物模型中能够诱导针对未修饰肿瘤细胞的免疫反应。为治疗人类患者，研发了一种由自体肿瘤细胞和分泌白细胞介素 -2（IL-2）的同种异基因成纤维细胞组成的疫苗。自体肿瘤细胞从活检标本中分离得到。通过用人类IL-2和新霉素抗性（Neo(r)）的表达构建体进行阳离子脂质转染，获得了一种永生化人类成纤维细胞系的克隆（KMST 6.14），该克隆每10(6)个细胞每24小时稳定分泌5290国际单位的IL-2。15例难治性恶性肿瘤患者在一项I期临床试验中接受了3 - 4次照射后的KMST6.14和自体肿瘤细胞注射。仅在接种部位以及注射照射后的肿瘤细胞后，出现了逐渐增强的短暂炎症反应且无全身毒性（p < 0.05）。这些部位含有密集浸润的CD3 + T细胞，其中CD4 + 辅助性T细胞的数量超过CD8 + 细胞毒性T细胞（CTL）。从2例恶性黑色素瘤患者而非肾细胞癌患者的接种部位分离出的CD8 + T细胞系在体外对自体肿瘤细胞表现出显著的裂解活性。从2例肾细胞癌患者中的1例接种部位建立的CD8 + T细胞克隆优先裂解自体和部分匹配的同种异基因肾细胞癌细胞。总之，由IL-2基因转染的同种异基因成纤维细胞和自体肿瘤细胞组成的疫苗能够在体内增强特异性抗肿瘤T细胞反应且无主要副作用。恶性黑色素瘤和肾细胞癌似乎是未来治疗试验中测试类似方法的有前景的对象。
