Eibl B, Ebner S, Duba C, Böck G, Romani N, Erdel M, Gächter A, Niederwieser D, Schuler G
Department of Internal Medicine, University of Innsbruck, Austria.
Genes Chromosomes Cancer. 1997 Nov;20(3):215-23. doi: 10.1002/(sici)1098-2264(199711)20:3<215::aid-gcc1>3.0.co;2-5.
Dendritic cells (DC) are professional antigen-presenting cells specialized in the initiation of primary immune responses. We were interested to know whether mature DC can be grown in vitro from peripheral blood mononuclear cells (PBMC) of patients with chronic myelogenous leukemia (CML), and whether they carry the Philadelphia (Ph) translocation. Using a method recently described, DC were generated from PBMC precursors of 12 patients with CML using GM-CSF, IL-4, and monocyte-conditioned medium. DC exhibited the typical morphology with thin cytoplasmatic processes and expressed high levels of MHC class II, CD86, and CD83 typical for mature DC. After sorting with the monoclonal antibody CD83, a cell population of more than 95% CD83 positive cells was obtained. The presence of the Ph translocation was analyzed in these cells, in PBMC, lymphoblastoid cell lines (LCL), and in phytohemagglutinin (PHA)-induced T blasts from the same patients by fluorescence in situ hybridization (FISH). In contrast to all other cells analyzed, the vast majority of DC (95.9 +/- 0.7%) displayed the Ph translocation, irrespective of disease stage or therapy. PBMC were predominantly positive for the Ph chromosome (67.6 +/- 7.3%), whereas only 11.4 +/- 1% of the B cells and 4.4 +/- 1.1% of the PHA blasts carried the Ph translocation. Using such leukemic DC as antigen-presenting cells, a primary CML-directed cytotoxic immune response in vitro was obtained, as shown by the specific recognition of Ph chromosome positive cells. We conclude that DC can be generated from blood progenitors of CML patients in vitro and exhibit, to a large extent, the Ph translocation. Such DC, which in a preliminary experiment have been able to induce a primary CML-directed cytotoxic immune response in vitro, might be ideal candidates for adoptive immunotherapy either by direct transfer of DC for in vivo generation of a T-cell response or by in vitro generation of CML-specific cytotoxic autologous or HLA-matched normal T-cell clones for use in vivo.
树突状细胞（DC）是专门负责启动初始免疫反应的专职抗原呈递细胞。我们想了解慢性髓性白血病（CML）患者外周血单个核细胞（PBMC）能否在体外培养出成熟的DC，以及这些DC是否携带费城染色体（Ph）易位。采用最近描述的一种方法，使用GM-CSF、IL-4和单核细胞条件培养基，从12例CML患者的PBMC前体细胞中生成DC。DC呈现出具有细细胞质突起的典型形态，并表达高水平的MHC II类分子、CD86和CD83，这些都是成熟DC的典型特征。用单克隆抗体CD83分选后，获得了一个CD83阳性细胞比例超过95%的细胞群体。通过荧光原位杂交（FISH）分析了这些细胞、PBMC、淋巴母细胞系（LCL）以及来自同一患者的植物血凝素（PHA）诱导的T母细胞中Ph易位的存在情况。与所有其他分析的细胞不同，绝大多数DC（95.9±0.7%）显示出Ph易位，与疾病阶段或治疗无关。PBMC中Ph染色体大多呈阳性（67.6±7.3%），而只有11.4±1%的B细胞和4.4±1.1%的PHA母细胞携带Ph易位。以这种白血病DC作为抗原呈递细胞，体外获得了针对原发性CML的细胞毒性免疫反应，这通过对Ph染色体阳性细胞的特异性识别得以证明。我们得出结论，DC可在体外从CML患者的血液祖细胞中生成，并且在很大程度上呈现Ph易位。这种DC在初步实验中已能够在体外诱导针对原发性CML的细胞毒性免疫反应，无论是通过直接转移DC以在体内产生T细胞反应，还是通过体外生成CML特异性细胞毒性自体或HLA匹配的正常T细胞克隆以供体内使用，都可能是过继性免疫治疗的理想候选者。
