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目的:探讨冠状动脉严重狭窄冠心病患者冠状动脉血树突状细胞(dendritic cells,DC)的数量、表型、功能状态与冠状动脉侧支循环的关系。方法:收集40例冠状动脉严重狭窄冠心病患者,根据冠状动脉侧支循环(coronary collateral circulation,CCC)形成情况分为CCC形成组(A组,n=22)和无CCC形成组(B组,n=18)。采用密度梯度离心法分离冠状动脉血单个核细胞,行DC体外培养与扩增,观察DC形态,检测收获细胞总数和DC数量;采用流式细胞仪检测DC表型与平均荧光强度(mean fluorescenceintensity,MFI),采用同种异体混合淋巴细胞反应检测DC刺激T淋巴细胞增殖能力,计算刺激指数(stim-ulation index,SI)。结果:(1)冠状动脉血分离单个核细胞后成功培养出典型DC,两组DC形态上无差异;(2)A组收获细胞数为(3.95±1.41)×106,B组收获细胞数为(2.76±0.92)×106,A组收获细胞数显著增多,差异有统计学意义(P=0.003);(3)A组DC数为(1.54±0.96)×106,B组DC数为(0.99±0.46)×106,A组DC数明显增多,差异有统计学意义(P=0.033);(4)两组CD1a阳性细胞比例;MFI,CD1a与CD80,CD83,CD86双阳性细胞比例及MFI差异均无统计学意义(P>0.05);(5)A组SI为4.96±2.30,B组SI为2.66±1.04,A组SI显著增高,差异有统计学意义(P=0.0003)。结论:在冠状动脉严重狭窄的冠心病患者中,冠状动脉侧支形成患者DC的数量显著增加,刺激T淋巴细胞增殖的能力增强。
Objective: To investigate the relationship between the number, phenotype and function of coronary artery dendritic cells (DCs) and coronary collateral circulation in patients with severe coronary artery stenosis. Methods: Forty patients with severe coronary artery stenosis and coronary artery disease were enrolled in this study. According to the formation of coronary collateral circulation (CCC), CCC group (group A, n = 22) and CCC group (group B, n = 18). The density of coronary artery blood mononuclear cells were isolated by density gradient centrifugation, and the DCs were cultured and expanded in vitro. The morphology of DCs was observed, the total number of DCs and the number of DCs were detected. The phenotypes of DCs were analyzed by flow cytometry, mean fluorescence intensity (mean fluorescence intensity, MFI). Allogeneic mixed lymphocyte reaction was used to detect the ability of DCs to stimulate the proliferation of T lymphocytes. The stimulatory index (SI) was calculated. (2) The number of cells harvested in group A was (3.95 ± 1.41) × 106, and the number of harvested cells in group B was (2.76 ± 0.92) × 106, the number of harvested cells in group A was significantly increased (P = 0.003); (3) The number of DC in group A was (1.54 ± 0.96) × 106; the number of DC in group B was ± 0.46) × 106, the number of DCs in group A increased significantly (P = 0.033); (4) The proportion of CD1a positive cells in both groups and the difference of MFI, CD1a and CD80, CD83, CD86 double positive cells and MFI (5) The SI in group A was 4.96 ± 2.30, and the SI in group B was 2.66 ± 1.04. SI in group A was significantly higher than that in group A (P = 0.0003). Conclusion: In patients with severe coronary artery stenosis, the number of DCs in coronary collateral formation is significantly increased, and the ability of stimulating T lymphocyte proliferation is enhanced.