University Cell Immunotherapy Centre, Masaryk University, Kamenice 5, Brno, Czech Republic.
Br J Haematol. 2010 Mar;148(6):859-67. doi: 10.1111/j.1365-2141.2009.08034.x. Epub 2010 Jan 13.
Despite novel treatment strategies, multiple myeloma (MM) remains an incurable disease with low immunogenicity and multiple immune defects. We developed an ex vivo strategy for inducing myeloma-specific cytotoxic T lymphocytes (CTLs) and demonstrate the possibility of identification and long-term in vivo monitoring of individual myeloma-specific T-cell clones using the most sensitive clonotypic assay that is able to detect low frequencies of T-cell clones (1 clonotypic cell in 10(6) cells). Ten patients with MM were examined for the presence of tumour-reactive T cells using dendritic cells loaded with autologous tumour cells. All patients had detectable myeloma-reactive T cells in vitro. Expanded myeloma-reactive T cells demonstrated specific cytotoxic effects against autologous tumour cells in vitro (median 39.6% at an effector:target ratio of 40:1). The clonality of myeloma-specific T cells was studied with a clonotypic assay, which demonstrated both oligoclonal and monoclonal populations of myeloma-specific T cells. CD8(+) CTLs were the most immunodominant myeloma-specific T-cell clones and clinical responses were closely associated with the in vivo expansion and long-term persistence of individual CD8(+) T-cell clones, usually at very low frequencies (10(-3)-10(-6)). We conclude that the clonotypic assay is the most sensitive tool for immunomonitoring of low-frequency T cells.
尽管有新的治疗策略，多发性骨髓瘤（MM）仍然是一种无法治愈的疾病，其免疫原性低，存在多种免疫缺陷。我们开发了一种诱导骨髓瘤特异性细胞毒性 T 淋巴细胞（CTL）的体外策略，并利用最敏感的能够检测低频率 T 细胞克隆的克隆型检测方法，证明了识别和长期体内监测个体骨髓瘤特异性 T 细胞克隆的可能性（在 10^6 个细胞中 1 个克隆型细胞）。对 10 名 MM 患者进行了使用负载自体肿瘤细胞的树突状细胞检测肿瘤反应性 T 细胞的检查。所有患者均在体外检测到骨髓瘤反应性 T 细胞。扩增的骨髓瘤反应性 T 细胞在体外对自体肿瘤细胞表现出特异性细胞毒性作用（效应物：靶比为 40:1 时中位数为 39.6%）。使用克隆型检测研究了骨髓瘤特异性 T 细胞的克隆性，该检测方法显示骨髓瘤特异性 T 细胞存在寡克隆和单克隆群体。CD8+CTL 是最具免疫优势的骨髓瘤特异性 T 细胞克隆，临床反应与个体 CD8+T 细胞克隆的体内扩增和长期持续存在密切相关，通常频率非常低（10^-3-10^-6）。我们得出结论，克隆型检测是监测低频 T 细胞的最敏感工具。
