Labarriere N, Pandolfino M C, Raingeard D, Le Guiner S, Diez E, Le Dréan E, Dreno B, Jotereau F
Institut de Biologie, INSERM U463 and Faculté des Sciences de Nantes, France.
Int J Cancer. 1998 Oct 5;78(2):209-15. doi: 10.1002/(sici)1097-0215(19981005)78:2<209::aid-ijc15>3.0.co;2-5.
Several tumor antigens have been described as candidates for immunotherapy. Our study compared HLA-A2-restricted epitopes from 5 antigens commonly expressed by melanomas, i.e., Melan-A/MART-1 peptides (26-35 and 27-35), tyrosinase (368-376), gp-100 (280-288), MAGE-3 (271-279) and NA17-A (1-10), for their relative capacity to promote the development of cytotoxic and cytokine-producing specific CD8+ lymphocytes within melanoma-invaded lymph nodes. We used short-term cultured melanoma-invaded lymph node lymphocytes (MILLs) and tested responses developed by these cells to peptide-pulsed TAP-deficient T2 cells. We measured both the lytic response developed by MILLs and the fraction of these cells that secreted interferon-gamma (IFN-gamma), as deduced from intracellular cytokine labeling. Reverse transcription-polymerase chain reaction (RT-PCR) was used to analyze the expression of the 5 antigens within melanoma-invaded lymph nodes. Melan-A/MART-1, tyrosinase and gp-100 peptides were recognized by MILLs derived, respectively, from 8 of 20, 5 of 19 and 4 of 20 melanoma-invaded lymph nodes expressing these antigens. Most MILLs specific for Melan-A/MART-1 and tyrosinase exhibited both lysis and IFN-gamma responses, whereas most of those specific for gp-100 developed only lysis. Weak lysis without IFN-gamma secretion was developed against NA17-A and MAGE-3 peptides by MILLs from, respectively, 3 of 9 and 2 of 14 lymph nodes expressing these antigens. Our data show a prevalence of both cytotoxic and IFN-gamma-secreting effector T cells specific for differentiation antigens within HLA-A2 melanoma-invaded lymph nodes, which makes these antigens attractive targets for specific immunotherapy.
几种肿瘤抗原已被描述为免疫治疗的候选对象。我们的研究比较了黑色素瘤通常表达的5种抗原的HLA - A2限制性表位，即黑色素瘤抗原A/MART - 1肽（26 - 35和27 - 35）、酪氨酸酶（368 - 376）、糖蛋白100（280 - 288）、黑色素瘤相关抗原3（MAGE - 3）（271 - 279）和NA17 - A（1 - 10），比较它们在促进黑色素瘤浸润淋巴结内细胞毒性和产生细胞因子的特异性CD8 +淋巴细胞发育方面的相对能力。我们使用短期培养的黑色素瘤浸润淋巴结淋巴细胞（MILLs），并测试这些细胞对肽脉冲处理的TAP缺陷型T2细胞产生的反应。我们测量了MILLs产生的裂解反应以及这些细胞中分泌干扰素 - γ（IFN - γ）的比例，这是通过细胞内细胞因子标记推断出来的。逆转录 - 聚合酶链反应（RT - PCR）用于分析黑色素瘤浸润淋巴结内这5种抗原的表达。黑色素瘤抗原A/MART - 1、酪氨酸酶和糖蛋白100肽分别被来自表达这些抗原的20个黑色素瘤浸润淋巴结中的8个、19个中的5个和20个中的4个的MILLs识别。大多数对黑色素瘤抗原A/MART - 1和酪氨酸酶特异的MILLs表现出裂解和IFN - γ反应，而大多数对糖蛋白100特异的MILLs仅产生裂解反应。来自分别表达这些抗原的9个淋巴结中的3个和14个淋巴结中的2个的MILLs对NA17 - A和MAGE - 3肽产生了无IFN - γ分泌的微弱裂解反应。我们的数据显示，在HLA - A2黑色素瘤浸润淋巴结内，对分化抗原特异的细胞毒性和分泌IFN - γ的效应T细胞普遍存在，这使得这些抗原成为特异性免疫治疗的有吸引力的靶点。
