Analysis of dendritic cells from patients with hematologic neoplasms

我们的研究对象是确诊为急性髓细胞白血病(AML)、非霍奇金淋巴瘤(NHL)、霍奇金病(HD)和骨髓增生异常综合征(MDS)的患者。除急性髓系白血病患者持续完全缓解外，其余患者均于确诊时采集外周血。采用Ficoll密度离心法、T玫瑰花环法、贴片法和甲硝胺密度离心法从外周血中分离纯化树突状细胞。DC功能通过其刺激同种异体T细胞的能力来评估。用双色流式细胞术检测CD83阳性DC表面黏附分子(CD11a、CD54、CD58)、共刺激分子(CD80、CD86、CD40)和人类白细胞抗原(DR、DP、DQ)的表达。NHL和HD患者的DC功能因患者不同而不同，但与正常志愿者(NV)相比，大多数患者DC功能降低。NV与NHL或HD患者DC表面功能重要分子的表达无显著差异。AML患者DC功能至少与NV相当。大多数MDS患者的DC功能与NV相似。荧光原位杂交分析发现，部分DC来源于MDS患者的异常克隆。

Our subjects were patients who had been diagnosed with acute mayeloid leukemia (AML), non-Hodgkin's lymphoma (NHL), Hodgkin's disease (HD) and myelodysplastic syndromes (MDS). Peripheral blood (PB) was obtained from the patients at diagnosis, except for the AML patients, who were in continued complete remission. Dendritic cells (DC) were purified from PB by Ficoll density centrifugation, T rosetting, dish adherence and metrizamide density centrifugation. DC function was assessed by their capacity to stimulate allogeneic T cells. Expression of functionally important molecules on CD83-positive DC, I.e., adhesion molecules (CD11a, CD54, CD58), co-stimulatory molecules (CD80, CD86, CD40) and HLA (DR, DP, DQ), was analyzed by two-color flow cytometry. The function of DC from NHL and HD patients varied among patients, but was reduced in most patients when compared with the DC function of normal volunteers (NV). Expression of functionally important molecules on DC did not differ significantly between NV and patients with NHL or HD. DC function of AML patients was at least equal to that of NV. The DC function of most MDS patients was similar to that of NV. Some DC were found to be derived from an abnormal clone in MDS patients when analyzed by fluorescence in situ hybridization.