Arienti F, Sulé-Suso J, Belli F, Mascheroni L, Rivoltini L, Melani C, Maio M, Cascinelli N, Colombo M P, Parmiani G
Division of Experimental Oncology D, Istituto Nazionale Tumori, Milan, Italy.
Hum Gene Ther. 1996 Oct 20;7(16):1955-63. doi: 10.1089/hum.1996.7.16-1955.
We have immunized advanced melanoma patients with a HLA-A2-compatible human melanoma line genetically modified to release interleukin-2 (IL-2), to elicit or increase a T cell-mediated anti-melanoma response that may affect distant lesions. Twelve stage-IV patients were injected subcutaneously at days 1, 13, 26, and 55 with IL-2 gene-transduced and irradiated melanoma cells at doses of 5 or 15 x 10(7) cells. Both local and systemic toxicities were mild, consisting of transient erythema at the vaccination site; fever occurred in a minority of patients. Three mixed responses were recorded. Seven patients were evaluable for immunological studies. Mixed tumor-lymphocyte cultures carried out with different allogeneic HLA-A2-matched melanoma lines as stimulators and targets revealed an increase in the MHC-unrestricted, but no changes in the MHC-restricted, cytotoxicity in peripheral blood lymphocytes (PBL) obtained after vaccination as compared with those obtained before vaccination. Increased recognition of the tyrosinase 368-376 peptide occurred in post-vaccination PBL of one patient, whereas a weak increase in recognition of the gp100 280-288 peptide was detectable in another patient; these 2 patients also recognized the gp100 457-466 peptide. After in vitro, stimulation with the only available autologous melanoma line, CD4+ cells with autologous tumor-specific cytotoxicity and ability to release interferon-gamma (IFN-gamma) were found in post- but not in pre-vaccination PBL. In the same patient, as well as in another patient, limiting dilution analysis showed that vaccination resulted in an increased frequency of melanoma-specific cytotoxic T lymphocyte (CTL) precursors. These results indicate that vaccination with cells releasing IL-2 locally can expand a T cell response against antigen(s) of autologous, untransduced tumor, although this response occurred in a minority of the melanoma patients studied.
我们用一种经基因改造可释放白细胞介素-2（IL-2）的HLA-A2兼容人黑色素瘤细胞系对晚期黑色素瘤患者进行免疫接种，以引发或增强T细胞介导的抗黑色素瘤反应，该反应可能会影响远处病灶。12例IV期患者在第1、13、26和55天皮下注射剂量为5或15×10⁷个细胞的IL-2基因转导且经辐照的黑色素瘤细胞。局部和全身毒性均较轻，包括接种部位的短暂红斑；少数患者出现发热。记录到3例混合反应。7例患者可进行免疫学研究。用不同的同种异体HLA-A2匹配黑色素瘤细胞系作为刺激物和靶细胞进行的混合肿瘤-淋巴细胞培养显示，与接种前相比，接种后外周血淋巴细胞（PBL）中主要组织相容性复合体（MHC）非限制性细胞毒性增加，但MHC限制性细胞毒性无变化。一名患者接种后的PBL中对酪氨酸酶368 - 376肽的识别增加，而在另一名患者中可检测到对gp100 280 - 288肽的识别略有增加；这两名患者也识别gp100 457 - 466肽。在体外，用唯一可用的自体黑色素瘤细胞系刺激后，接种后而非接种前的PBL中发现具有自体肿瘤特异性细胞毒性和释放γ干扰素（IFN-γ）能力的CD4⁺细胞。在同一名患者以及另一名患者中，有限稀释分析表明接种导致黑色素瘤特异性细胞毒性T淋巴细胞（CTL）前体频率增加。这些结果表明，局部注射释放IL-2的细胞进行接种可扩大针对自体未转导肿瘤抗原的T细胞反应，尽管这种反应仅在少数研究的黑色素瘤患者中出现。
