Das Gupta T K, Cohen E P, Richards J M
Hum Gene Ther. 1997 Sep 20;8(14):1701-14. doi: 10.1089/hum.1997.8.14-1701.
A cell line (UISO-H-MEL-2) was established from the neoplastic cells of a patient with malignant melanoma during the natural course of the patient's treatment. The melanoma cells express defined MHC Class I histocompatibility determinants including determinants specified by the HLA-A2 Class I allele, along with a common melanoma-associated T-cell epitope derived from the tyrosinase gene. The gene for human interleukin-2 (IL-2) was transduced into the cells with a provirus (pZipNeoSVIL-2), packaged in GP + envAM12 cells. Integration of the IL-2 gene into genomic DNA of the transduced cells and its expression were established. The IL-2-secreting cell line (UISO-H-MEL-2-IL-2) was found to be free of recombinant retroviruses and other infectious agents. The IL-2-secreting cells will be subjected to 5000 rads X-irradiation and administered to 12 informed patients with metastatic malignant melanoma in a Phase I toxicity study. The dose of X-irradiation was sufficient to inactivate one hundred percent of the cells, but insufficient to completely inhibit IL-2 synthesis during a fourteen-day period of analysis. Patients who have failed all standard forms of treatment will become eligible for inclusion in the study if they develop metastatic melanoma, and if their tumor cells express products of the tyrosinase gene. The patients will differ with the cellular immunogen at no less than three of six MHC Class I alleles, but will share identity at the HLA-A2 Class I allele. The patient's antimelanoma immune response to the injected cells will be determined by both in vivo and in vitro parameters. Background studies performed in inbred mice indicate that X-irradiated IL-2-secreting cells that express both melanoma-associated antigens and allogeneic Class I histocompatibility antigens are more antigenic in terms of their capacity to induce an antimelanoma response than X-irradiated IL-2-secreting melanoma cells. Of significance for the future potential of this form of therapy in melanoma patients, the period of survival of mice was established melanoma treated with the IL-2-secreting allogeneic cells was significantly (P < 0.001) longer than that of untreated animals, or animals treated with X-irradiated melanoma cells. An analogous protocol was reviewed and approved by the Recombinant DNA Advisory Committee of the National Institutes of Health.
在一名恶性黑色素瘤患者治疗的自然病程中，从其肿瘤细胞建立了一个细胞系（UISO-H-MEL-2）。黑色素瘤细胞表达特定的MHC I类组织相容性决定簇，包括由HLA-A2 I类等位基因指定的决定簇，以及一个源自酪氨酸酶基因的常见黑色素瘤相关T细胞表位。用前病毒（pZipNeoSVIL-2）将人白细胞介素-2（IL-2）基因转导到细胞中，并包装在GP + envAM12细胞中。确定了IL-2基因整合到转导细胞的基因组DNA中及其表达情况。发现分泌IL-2的细胞系（UISO-H-MEL-2-IL-2）不含重组逆转录病毒和其他感染因子。在一项I期毒性研究中，将分泌IL-2的细胞进行5000拉德的X射线照射后，给予12名知情的转移性恶性黑色素瘤患者。X射线照射剂量足以使100%的细胞失活，但不足以在为期14天的分析期内完全抑制IL-2的合成。如果所有标准治疗形式均失败的患者发生转移性黑色素瘤，且其肿瘤细胞表达酪氨酸酶基因的产物，则有资格纳入该研究。患者在六个MHC I类等位基因中至少三个上与细胞免疫原不同，但在HLA-A2 I类等位基因上相同。将通过体内和体外参数确定患者对注射细胞的抗黑色素瘤免疫反应。在近交系小鼠中进行的背景研究表明，表达黑色素瘤相关抗原和同种异体I类组织相容性抗原的经X射线照射的分泌IL-2的细胞，就其诱导抗黑色素瘤反应的能力而言，比经X射线照射的分泌IL-2的黑色素瘤细胞更具抗原性。对于这种治疗形式在黑色素瘤患者中的未来潜力具有重要意义的是，用分泌IL-2的同种异体细胞治疗的黑色素瘤小鼠的存活期明显（P < 0.001）长于未治疗的动物或用经X射线照射的黑色素瘤细胞治疗的动物。一项类似的方案已由美国国立卫生研究院的重组DNA咨询委员会审查并批准。
