Romero P, Dunbar P R, Valmori D, Pittet M, Ogg G S, Rimoldi D, Chen J L, Liénard D, Cerottini J C, Cerundolo V
Division of Clinical Onco-Immunology, Ludwig Institute for Cancer Research, Lausanne Branch, Lausanne, Swizerland.
J Exp Med. 1998 Nov 2;188(9):1641-50. doi: 10.1084/jem.188.9.1641.
Characterization of cytolytic T lymphocyte (CTL) responses to tumor antigens has been impeded by a lack of direct assays of CTL activity. We have synthesized reagents ("tetramers") that specifically stain CTLs recognizing melanoma antigens. Tetramer staining of tumor-infiltrated lymph nodes ex vivo revealed high frequencies of tumor-specific CTLs which were antigen-experienced by surface phenotype. In vitro culture of lymph node cells with cytokines resulted in very large expansions of tumor-specific CTLs that were dependent on the presence of tumor cells in the lymph nodes. Tetramer-guided sorting by flow cytometer allowed isolation of melanoma-specific CTLs and confirmation of their specificity and their ability to lyse autologous tumor cells. Our results demonstrate the value of these novel reagents for monitoring tumor-specific CTL responses and for generating CTLs for adoptive immunotherapy. These data also indicate that strong CTL responses to melanoma often occur in vivo, and that the reactive CTLs have substantial proliferative and tumoricidal potential.
缺乏直接检测细胞毒性T淋巴细胞（CTL）活性的方法阻碍了对CTL针对肿瘤抗原反应的特性研究。我们合成了能特异性标记识别黑色素瘤抗原的CTL的试剂（“四聚体”）。对肿瘤浸润淋巴结进行体外四聚体染色显示，具有抗原接触表面表型的肿瘤特异性CTL频率很高。用细胞因子对淋巴结细胞进行体外培养，可使肿瘤特异性CTL大量扩增，这依赖于淋巴结中肿瘤细胞的存在。通过流式细胞仪进行四聚体引导分选，能够分离出黑色素瘤特异性CTL，并确认其特异性以及裂解自体肿瘤细胞的能力。我们的结果证明了这些新型试剂在监测肿瘤特异性CTL反应以及为过继性免疫治疗生成CTL方面的价值。这些数据还表明，对黑色素瘤的强烈CTL反应常在体内发生，并且反应性CTL具有显著的增殖和杀瘤潜力。
