Arienti F, Sulé-Suso J, Melani C, Maccalli C, Belli F, Illeni M T, Anichini A, Cascinelli N, Colombo M P, Parmiani G
Hum Gene Ther. 1994 Sep;5(9):1139-50. doi: 10.1089/hum.1994.5.9-1139.
Two human melanoma lines were transduced by a retroviral vector with the gene of the human interleukin-2 (IL-2) and characterized for their immunological properties in comparison with the parental lines. Transduction resulted in the production of biologically active IL-2 in the average amounts of 2,282 and 2,336 pg/ml per 10(5) cells per 24 hr over 3 and 2 months by the Me14932/IL-2 and the Me1B6/IL-2 lines, respectively. Melanoma-transduced cells lost their tumorigenicity in nude mice. No major changes in the phenotype were observed in IL-2 gene-transduced lines. In fact, more than 90% of cells expressed class I and II(DR) HLA, adhesion molecules, integrins, and melanoma-associated antigens. Irradiation with 100-400 Gy, while inhibiting tumor cell growth in vitro, allowed the release of IL-2 by the transduced cells for at least 5 weeks. The two melanoma lines also maintained susceptibility to lysis by lymphokine-activated killer (LAK) cells and by a HLA-A2-restricted melanoma-specific cytotoxic T lymphocyte (CTL) clone recognizing the melanoma antigen (Melan-A). In a limiting dilution assay, transduced, but not parental melanoma lines unless added with an amount of IL-2 comparable to that released by the transduced cells, were able to expand both nonspecific and melanoma-specific CTL precursors from autologous peripheral blood lymphocytes (PBL). In mixed lymphocytes-tumor cultures, IL-2 gene-transduced melanoma cells stimulated the expansion of major histocompatibility complex (MHC)-unrestricted effectors from autologous PBL, and of CD3+ CD8+ MHC-restricted CTL from tumor-invaded lymph nodes. These results indicate that IL-2 gene transduction does not alter significantly the expression of the immunologically relevant molecules of human melanoma lines while increasing their ability to stimulate both specific and nonspecific lymphocyte responses. These lines will be of value in the vaccination of melanoma patients.
用携带人白细胞介素 -2（IL -2）基因的逆转录病毒载体转导两个人黑色素瘤细胞系，并与亲代细胞系相比，对其免疫特性进行表征。转导导致Me14932/IL -2和Me1B6/IL -2细胞系在3个月和2个月的时间里，每10⁵个细胞每24小时分别平均产生2282和2336 pg/ml的生物活性IL -2。转导的黑色素瘤细胞在裸鼠中失去了致瘤性。在IL -2基因转导的细胞系中未观察到表型的重大变化。事实上，超过90%的细胞表达I类和II类（DR）HLA、黏附分子、整合素以及黑色素瘤相关抗原。用100 - 400 Gy照射，在抑制体外肿瘤细胞生长的同时，使转导细胞释放IL -2至少持续5周。这两个黑色素瘤细胞系对淋巴因子激活的杀伤（LAK）细胞和识别黑色素瘤抗原（Melan -A）的HLA -A2限制性黑色素瘤特异性细胞毒性T淋巴细胞（CTL）克隆的裂解仍保持敏感性。在有限稀释试验中，转导的黑色素瘤细胞系能够从自体外周血淋巴细胞（PBL）中扩增非特异性和黑色素瘤特异性CTL前体，而亲代黑色素瘤细胞系除非添加与转导细胞释放量相当的IL -2则不能。在混合淋巴细胞 -肿瘤培养中，IL -2基因转导的黑色素瘤细胞刺激了来自自体PBL的主要组织相容性复合体（MHC）非限制性效应细胞以及来自肿瘤浸润淋巴结的CD3⁺CD8⁺MHC限制性CTL的扩增。这些结果表明，IL -2基因转导不会显著改变人黑色素瘤细胞系免疫相关分子的表达，同时增强其刺激特异性和非特异性淋巴细胞反应的能力。这些细胞系在黑色素瘤患者的疫苗接种中将具有价值。
