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目的 研究CD1a和CD83分子是否特异性地表达在乳腺癌患者外周血中培养的树突状细胞上。方法 采用密度梯度离心的方法,分离32例乳腺癌患者外周血中的单个核细胞,在6孔培养板上(106/ml,2ml/孔),用含10％热灭火的小牛血清和GM-CSF、IL-4、TNF-α细胞因子的RPMI1640培养基培养。(贴壁)2小时后,轻轻吸去非贴壁细胞并加入新鲜培养基。用标有荧光素的单抗标记培养细胞,在流式细胞仪上分析培养细胞,本研究中所用的单抗有异硫氰酸荧光素(FITC)标记的CD1a、CD3、CD19、CD40、CD80和藻红朊(PE)标记的CD3、CD19、CD83、CD86、MHCⅡ。在激光共聚焦显微镜下拍摄标记有荧光素的细胞。结果 该细胞表达特征性分子CD1a、CD4O、CD80、CD83、CD86,CD1a、CD83在CD3+T、CD19+B淋巴细胞上也表达,在激活的淋巴细胞上表达水平高于未激活的淋巴细胞,而且B淋巴细胞上的表达水平高于T淋巴细胞。结论 CD1a和CD83分子并非DC特有的标记,目前鉴定DC仍然要靠细胞形态、细胞表达CD1a、CD83以及共刺激分子等综合因素来确定DC。
Objective To investigate whether CD1a and CD83 molecules are specifically expressed on dendritic cells cultured in the peripheral blood of breast cancer patients. Methods Peripheral blood mononuclear cells from 32 patients with breast cancer were isolated by density gradient centrifugation and cultured in 6-well plates (106 / ml, 2 ml / well) with 10% heat-killed calf serum and GM -CSF, IL-4, TNF-α cytokines in RPMI1640 medium. (Adherent) After 2 hours, gently pipette non-adherent cells and add fresh medium. The cultured cells were labeled with fluorescein-labeled monoclonal antibody and the cultured cells were analyzed by flow cytometry. The monoclonal antibodies used in this study were fluorescein isothiocyanate (FITC) -labeled CD1a, CD3, CD19, CD40, CD80 And phycoerythrin (PE) labeled CD3, CD19, CD83, CD86, MHCII. Cells labeled with fluorescein were photographed under a laser confocal microscope. Results The expressed CD1a, CD4O, CD80, CD83, CD86, CD1a and CD83 were also expressed on CD3 + T and CD19 + B lymphocytes. The expression of CD1a, CD80, CD83 and CD83 were higher in activated lymphocytes than in inactive lymphocytes, Moreover, the level of expression on B lymphocytes is higher than that on T lymphocytes. Conclusion The CD1a and CD83 molecules are not unique markers of DC. Currently, DCs are still identified by the cell morphology, cells express CD1a, CD83 and costimulatory molecules to determine DC.