Smit W M, Rijnbeek M, van Bergen C A, Fibbe W E, Willemze R, Falkenburg J H
Department of Hematology, Leiden University Medical Center, 2300 RC Leiden, The Netherlands.
Proc Natl Acad Sci U S A. 1998 Aug 18;95(17):10152-7. doi: 10.1073/pnas.95.17.10152.
Adoptive immunotherapy with donor lymphocyte infusions (DLI) is an effective treatment for relapsed chronic myeloid leukemia (CML) after allogeneic stem cell transplantation. To identify the effector and target cell populations responsible for the elimination of the leukemic cells in vivo we developed an assay to measure the frequency of T lymphocyte precursor cells capable of suppressing leukemic progenitor cells. Target cells in this assay were CML cells that were cultured in the presence of stem cell factor, interleukin 3, granulocyte-macrophage colony-stimulating factor, granulocyte colony-stimulating factor, and erythropoietin. [3H]thymidine incorporation at day 7 represented the proliferation of the progeny of the CD34(+) CML progenitor cells, and not of the more mature CD34(-) CML cells. Effector cells were mononuclear cells, which were used in a limiting dilution analysis to measure the frequencies of CML progenitor cell-inhibitory lymphocyte precursors (PCILp) in peripheral blood of seven patients before and after DLI for relapsed CML. In the six patients who entered complete remission, a 5- to 100-fold increase of PCILp was found during the clinical response. In the patient with resistant relapse the frequency of PCILp was <10 per ml before and after DLI. Leukemia-reactive helper T lymphocyte precursor frequencies remained unchanged after DLI. A significant increase in cytotoxic T lymphocyte precursor frequency against more mature leukemic cells was found in only two responding patients. These results indicate that T cells specifically directed against CD34(+) CML progenitor cells mediate the antileukemic effect of DLI.
采用供体淋巴细胞输注(DLI)进行过继性免疫治疗是异基因干细胞移植后复发的慢性粒细胞白血病(CML)的一种有效治疗方法。为了确定体内负责清除白血病细胞的效应细胞和靶细胞群体,我们开发了一种检测方法来测量能够抑制白血病祖细胞的T淋巴细胞前体细胞的频率。该检测中的靶细胞是在干细胞因子、白细胞介素3、粒细胞-巨噬细胞集落刺激因子、粒细胞集落刺激因子和促红细胞生成素存在的情况下培养的CML细胞。第7天的[3H]胸苷掺入代表CD34(+) CML祖细胞后代的增殖,而不是更成熟的CD34(-) CML细胞的增殖。效应细胞是单核细胞,用于有限稀释分析,以测量7例复发CML患者在DLI前后外周血中CML祖细胞抑制性淋巴细胞前体细胞(PCILp)的频率。在进入完全缓解的6例患者中,临床反应期间PCILp增加了5至100倍。在耐药复发的患者中,DLI前后PCILp的频率均<10/ml。DLI后白血病反应性辅助性T淋巴细胞前体频率保持不变。仅在2例有反应的患者中发现针对更成熟白血病细胞的细胞毒性T淋巴细胞前体频率显著增加。这些结果表明,特异性针对CD34(+) CML祖细胞的T细胞介导了DLI的抗白血病作用。