Michalek Jaroslav, Kocak Ivo, Fait Vuk, Zaloudik Jan, Hajek Roman
Cell Immunotherapy Center, Masaryk University, Brno, Czech Republic.
J Immunol. 2007 Jun 1;178(11):6789-95. doi: 10.4049/jimmunol.178.11.6789.
We investigated the presence of individual melanoma-specific T cell clones in patients with metastatic melanoma. Ten patients were examined for the presence of melanoma-reactive T cells using dendritic cells loaded with autologous tumor cells. Their specificity was tested using nonradioactive cytotoxicity test. Individual immunodominant T cell clones were identified by the clonotypic assay that combines in vitro cell culture, immunomagnetic sorting of activated IFN-gamma(+) T cells, TCRbeta locus-anchored RT-PCR, and clonotypic quantitative PCR. All patients had detectable melanoma-reactive T cells in vitro. Expanded melanoma-reactive T cells demonstrated specific cytotoxic effect against autologous tumor cells in vitro. Three patients experienced objective responses, and their clinical responses were closely associated with the in vivo expansion and long-term persistence of individual CD8(+) T cell clones with frequencies of 10(-6) to 10(-3) of all circulating CD8(+) T cells. Five patients with progressive disease experienced no or temporary presence of circulating melanoma-reactive T cell clones. Thus, circulating immunodominant CD8(+) T cell clones closely correlate with clinical outcome in patients with metastatic melanoma.
我们研究了转移性黑色素瘤患者体内是否存在单个黑色素瘤特异性T细胞克隆。使用负载自体肿瘤细胞的树突状细胞对10例患者进行了黑色素瘤反应性T细胞检测。采用非放射性细胞毒性试验检测其特异性。通过克隆型分析鉴定单个免疫优势T细胞克隆，该分析结合了体外细胞培养、活化的IFN-γ(+) T细胞的免疫磁珠分选、TCRβ基因座锚定的RT-PCR和克隆型定量PCR。所有患者在体外均可检测到黑色素瘤反应性T细胞。体外扩增的黑色素瘤反应性T细胞对自体肿瘤细胞具有特异性细胞毒性作用。3例患者出现客观反应，其临床反应与体内单个CD8(+) T细胞克隆的扩增和长期持续存在密切相关，这些克隆在所有循环CD8(+) T细胞中的频率为10(-6)至10(-3)。5例疾病进展患者未出现或仅短暂出现循环黑色素瘤反应性T细胞克隆。因此，循环免疫优势CD8(+) T细胞克隆与转移性黑色素瘤患者的临床结局密切相关。
