Schuster Friedhelm R, Buhmann Raymund, Reuther Susanne, Hubner Bernd, Grabrucker Christine, Liepert Anja, Reibke Roland, Lichtner Peter, Yang Ting, Kroell Tanja, Kolb Hans-Jochem, Borkhardt Arndt, Schmetzer Helga
Department of Pediatric Oncology, Hematology and Immunology, Heinrich Heine University Medical Center, Dusseldorf, Germany.
Cancer Genomics Proteomics. 2008 Sep-Oct;5(5):275-86.
Recently it was shown that myeloid leukemic cells can be induced to differentiate into leukemia-derived dendritic cells (DCleu), regaining the stimulatory capacity of professional DCs while presenting the leukemic antigen repertoire. But so far, the induced antileukemic T-cell responses have varied in specificity and efficacy, or have even mediated opposite effects. In an attempt to further characterize the DC/DCleu induced T-cell response pattern, immunoscope spectratyping, a novel and powerful tool to detect T-cell receptor (TCR) rearrangements was used in combination with functional flow cytometry and non-radioactive fluorolysis assays. Human leucocyte antigen (HLA) matched donor T-cells were repeatedly stimulated, either with leukemic blasts (French-American-British, FAB M4eo) or the corresponding blast-derived DCs. Functional comparison revealed no significant difference in their T-cell stimulatory capacity, while the DC/DCleu fraction favored T-cells with a higher lytic activity, comprising a higher proportion of T-memory CD45R0+ cells. Stimulation with blasts and DC/DCleu induced a similar TCR restriction pattern, while stimulation with DC/DCleu favored the CD4 T-cell subset and seemed to cause a higher grade of restriction. In conclusion, a combined strategy using spectratyping with functional tests might not only provide useful information about the specificity and efficacy of the induced T-cell response, but also pave the way to gain effective T-cell clones for therapeutic use.
最近有研究表明，髓系白血病细胞可被诱导分化为白血病衍生树突状细胞（DCleu），在呈现白血病抗原库的同时恢复专职树突状细胞的刺激能力。但到目前为止，诱导产生的抗白血病T细胞反应在特异性和效力方面存在差异，甚至产生相反的效果。为了进一步表征DC/DCleu诱导的T细胞反应模式，将免疫荧光光谱分型（一种检测T细胞受体（TCR）重排的新颖且强大的工具）与功能流式细胞术和非放射性荧光溶解分析相结合使用。用人白细胞抗原（HLA）匹配的供体T细胞反复进行刺激，刺激物为白血病母细胞（法美英协作组，FAB M4eo）或相应的母细胞衍生树突状细胞。功能比较显示它们的T细胞刺激能力没有显著差异，而DC/DCleu部分更有利于具有较高裂解活性的T细胞，其中包含更高比例的T记忆CD45R0+细胞。用母细胞和DC/DCleu刺激诱导出相似的TCR限制性模式，而用DC/DCleu刺激更有利于CD4 T细胞亚群，并且似乎导致更高程度的限制性。总之，将光谱分型与功能测试相结合的策略不仅可能提供有关诱导的T细胞反应的特异性和效力的有用信息，而且还为获得用于治疗的有效T细胞克隆铺平道路。
