Savary C A, Grazziutti M L, Melichar B, Przepiorka D, Freedman R S, Cowart R E, Cohen D M, Anaissie E J, Woodside D G, McIntyre B W, Pierson D L, Pellis N R, Rex J H
Department of Surgical Oncology, University of Texas M.D. Anderson Cancer Center, Houston 77030, USA.
Cancer Immunol Immunother. 1998 Jan;45(5):234-40. doi: 10.1007/s002620050438.
We studied the potential of multidimensional flow cytometry to evaluate the frequency and maturation/activation status of dendritic cells in minimally manipulated peripheral blood mononuclear cell preparations (i.e., only separated on Ficoll-Hypaque) of normal donors and cancer patients. A rare subset of HLA-DR+ leukocytes (less than 1% mononuclear cells) was detected in blood of normal donors that displayed all the features of dendritic cells: these cells had high forward-light-scatter characteristics and coexpressed CD4, CD86 and CD54 surface antigens, but lacked the lineage-associated surface markers of T cells, B cells, monocytes, granulocytes or NK i.e. they were CD3-, CD19-, CD20-, CD14-, CD11b-, CD16-, CD56-). These physical and phenotypic properties were virtually identical to those of immunomagnetically sorted leukocytes characterized as dendritic-cells on the basis of morphology, phenotype and high stimulatory activity in allogeneic mixed-lymphocyte cultures. Using this flow-cytometric approach we observed that the frequency of dendritic cell-like cells in peripheral blood mononuclear cell specimens of cancer patients receiving chemotherapy alone or those recovering from stem cell transplantation was significantly lower than that of normal individuals (mean +/- SE: 0.36 +/- 0.05%, 0.14 +/- 0.06%, and 0.75 +/- 0.04% respectively). Multidimensional flow-cytometric analysis of dendritic cells might represent an important new tool for assessing immunocompetence, and for monitoring the effects of therapeutic regimens on the immune system.
我们研究了多维流式细胞术评估正常供体和癌症患者经最少操作的外周血单核细胞制剂（即仅通过Ficoll-泛影葡胺分离）中树突状细胞频率及成熟/激活状态的潜力。在正常供体血液中检测到一小部分罕见的HLA-DR⁺白细胞（少于1%的单核细胞），其具备树突状细胞的所有特征：这些细胞具有高前向光散射特性，共表达CD4、CD86和CD54表面抗原，但缺乏T细胞、B细胞、单核细胞、粒细胞或NK细胞的谱系相关表面标志物，即它们CD3⁻、CD19⁻、CD20⁻、CD14⁻、CD11b⁻、CD16⁻、CD56⁻。这些物理和表型特性与基于形态、表型及在同种异体混合淋巴细胞培养中的高刺激活性而被鉴定为树突状细胞的免疫磁珠分选白细胞几乎相同。使用这种流式细胞术方法，我们观察到仅接受化疗的癌症患者或从干细胞移植中恢复的患者外周血单核细胞标本中树突状细胞样细胞的频率显著低于正常个体（均值±标准误：分别为0.36±0.05%、0.14±0.06%和0.75±0.04%）。树突状细胞的多维流式细胞术分析可能是评估免疫能力以及监测治疗方案对免疫系统影响的一种重要新工具。
