Griffioen Marieke, Borghi Martina, Schrier Peter I, Osanto Susanne, Schadendorf Dirk
Department of Clinical Oncology, Leiden University Medical Center, The Netherlands.
Cancer Immunol Immunother. 2004 Aug;53(8):715-22. doi: 10.1007/s00262-004-0514-z. Epub 2004 Mar 3.
In melanoma patients, CD8+ cytotoxic T cells have been found recognizing self-proteins of which the expression is restricted to the melanocytic lineage. These melanocyte differentiation antigens are expressed in normal melanocytes as well as in 80-100% of primary and metastatic melanoma. In this report, six HLA-A0201-subtyped metastatic melanoma patients vaccinated with dendritic cells (DCs) pulsed with autologous tumor lysates and keyhole limpet hemocyanin (KLH) were screened for the presence of CD8+ T cells specific for three HLA-A0201-binding peptides derived from the melanosomal antigens MART-1/Melan-A, gp100, and tyrosinase. For this purpose, nonstimulated as well as in vitro peptide-stimulated peripheral blood mononuclear cells (PBMCs) were tested for peptide-specific IFN-gamma release by enzyme-linked immunosorbent spot (ELISpot) assays. Furthermore, expression of the melanosomal antigens MART-1/Melan-A, gp100, and tyrosinase in tumor lesions was analyzed by immunohistochemistry before and after vaccination. We also used the ELISpot technique to investigate whether KLH-specific T cells were induced and whether these cells released type 1 (IFN-gamma) and/or type 2 (IL-13) cytokines. Our data show induction of CD8+ T cells specific for the melanosomal peptides MART-1/Melan-A(27-35) or tyrosinase(1-9), as well as IFN-gamma-releasing KLH-specific T cells, in two of six vaccinated melanoma patients, but do not support an association between the induction of these T cells and clinical responses.
在黑色素瘤患者中，已发现CD8 + 细胞毒性T细胞能够识别其表达仅限于黑素细胞谱系的自身蛋白。这些黑素细胞分化抗原在正常黑素细胞以及80 - 100%的原发性和转移性黑色素瘤中均有表达。在本报告中，对6例HLA - A0201亚型的转移性黑色素瘤患者进行了筛查，这些患者接种了用自体肿瘤裂解物和钥孔戚血蓝蛋白（KLH）脉冲处理的树突状细胞（DC），以检测是否存在针对源自黑素体抗原MART - 1 / Melan - A、gp100和酪氨酸酶的三种HLA - A0201结合肽的CD8 + T细胞。为此，通过酶联免疫吸附斑点（ELISpot）测定法检测未刺激以及体外肽刺激的外周血单个核细胞（PBMC）中肽特异性干扰素 - γ的释放。此外，在接种疫苗前后，通过免疫组织化学分析肿瘤病变中黑素体抗原MART - 1 / Melan - A、gp100和酪氨酸酶的表达。我们还使用ELISpot技术研究是否诱导了KLH特异性T细胞，以及这些细胞是否释放1型（干扰素 - γ）和/或2型（白细胞介素 - 13）细胞因子。我们的数据显示，在6例接种疫苗的黑色素瘤患者中有2例诱导出了针对黑素体肽MART - 1 / Melan - A(27 - 35)或酪氨酸酶(1 - 9)的CD8 + T细胞以及释放干扰素 - γ的KLH特异性T细胞，但不支持这些T细胞的诱导与临床反应之间存在关联。
