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目的:探索可用于检测人外周血中CD4+CD25+Treg细胞的最佳标记物。方法:以52名健康人和47名非血液系统肿瘤患者为研究对象,采用多色免疫荧光素标记和多参数流式细胞术同时检测外周血中CD4+CD25high、CD4+CD25+FoxP3+和CD4+CD25+CD127lowT细胞,并以经典指标CD4+CD25high和CD4+CD25+FoxP3+为标准,分析和比较CD4+CD25+CD127low作为识别CD4+CD25+Treg细胞的可行性及其优势。结果:健康人和肿瘤患者外周血CD4+CD25high、CD4+CD25+FoxP3+和CD4+CD25+CD127lowT细胞占CD4+T细胞百分比分别为(1.769±0.682)和(2.958±1.392);(2.905±1.772)和(5.128±2.227)以及(5.396±1.306)和(7.175±2.565)。三者呈相同的趋势,即肿瘤患者组>健康人组,且三者之间呈显著正相关(P<0.05)。用两种不同标记法高纯度分选CD4+CD25high和CD4+CD25+CD127lowT细胞群后,FoxP3阳性率分别为90.9和92.7。结论:CD4+CD25+CD127low三标记法可以帮助识别CD4+CD25+Treg和部分激活的CD4+T细胞,提高CD4+CD25+Treg细胞的可检出数量,并且不影响细胞活性度,是反映CD4+CD25+Treg细胞更理想的指标。
Objective: To explore the best marker that can be used to detect CD4 + CD25 + Treg cells in human peripheral blood. Methods: Totally 52 healthy individuals and 47 non-hematological malignancies were enrolled in this study. Multicolor immunofluorescein labeling and multi-parameter flow cytometry were used to detect the levels of CD4 + CD25high, CD4 + CD25 + FoxP3 + and CD4 + CD25 + CD127lowT cells. The CD4 + CD25 + CD127low T cells were analyzed and compared with the classical indicators CD4 + CD25high and CD4 + CD25 + FoxP3 +. Results: The percentages of CD4 + CD25high, CD4 + CD25 + FoxP3 + and CD4 + CD25 + CD127lowT in peripheral blood of healthy people and cancer patients were (1.769 ± 0.682) and (2.958 ± 1.392), (2.905 ± 1.772) And (5.128 ± 2.227) and (5.396 ± 1.306) and (7.175 ± 2.565), respectively. The three trends were the same, that is, cancer patients> healthy people, and there was a significant positive correlation between the three (P <0.05). The positive rates of FoxP3 were 90.9 and 92.7 respectively after the high purity CD4 + CD25high and CD4 + CD25 + CD127lowT cell populations were sorted by two different labeling methods. Conclusion: The triple labeling of CD4 + CD25 + CD127low can help identify CD4 + CD25 + Treg and partially activated CD4 + T cells and increase the detectable number of CD4 + CD25 + Treg cells without affecting cell viability, + CD25 + Treg cells more ideal indicator.