CD4^+CD25^+CD127^(dim/-) Tregs细胞在乳腺癌患者外周血中表达及意义

Expression and significance of CD4^+CD25^+CD127^(dim/-) Treg cells in peripheral blood of breast cancer patients

目的研究CD4+CD25+CD127dim/-调节性T(Treg)细胞在乳癌患者和阴性对照组(纤维瘤)患者外周血中表达及意义;探索应用免疫磁珠分选技术分选人(乳腺癌患者)外周血CD4+CD25+CD127dim/-Treg细胞方法及体外培养方法。方法流式细胞术检测15例单纯乳腺癌患者和15例单纯乳腺纤维腺瘤患者外周血CD4+CD25+CD127adim/-Treg细胞表达水平,并行相关性分析;应用免疫磁珠分选技术分选乳腺癌患者外周血CD4+CD25+CD127dim/-Treg细胞;抗CD3包被,CD28,IL-2刺激体外增殖培养,流式细胞术检测纯度。结果乳腺癌患者外周血中CD4+CD25+CD127dim/-Treg细胞阳性率明显大于乳腺纤维瘤(P<0.05);乳腺癌患者外周血免疫磁珠分选CD4+CD25+CD127dim/-Treg,细胞计数板计数细胞量为4×105个,细胞流式检测阳性率为94.16%,体外增殖培养5 d后细胞计数为1×106个,流式检测CD4+CD25+CD127dim/-Treg细胞阳性率为90.09%。结论CD4+CD25+CD127dim/-Tregs细胞可能是乳腺癌发生发展的重要因素之一,调节性T细胞可以作为区别乳腺良恶性肿瘤生物学行为具有重要参考价值的标记物;应用免疫磁珠分选技术分选,抗CD3包被,CD28,IL-2刺激体外增殖培养后可获理想纯度CD4+CD25+CD127dim/-Tregs细胞,为进一步研究乳腺癌患者外周血Tregs细胞的TCR CDR3分子特征奠定基础。

Objective： To research the expressions and significance of CD4＋CD25＋CD127dim/- regulatory T （Treg） cells in periph- eral blood of breast cancer patients and breast fibroadenoma patients （ negative control group）, explore the sorting method and in vitro culture method of CD4＋CD25＋CD127dim/-Tregs Treg cells in pel4pheral blood of breast cancer patients by immunomagnetic cell sorting technique. Methods： Flow eytometry was used to detect the expression levels of CD4＋CD25＋CD127dim/-Tregs cells in peripheral blood of 15 pure breast cancer patients and 15 pure breast fibroadenoma patients, then a correlation analysis was conducted; immunomagnetic ceil sorting technique was used to sort CIM＋ CD25 ~ CD127 dim/- Treg cells in peripheral blood of breast cancer patients; anti-CD3 package, in vitro proliferation and cul- ture by stimulating CD28 and IL-2 were used in this study, flow cytometry was used to detect the purity. Results： The positive rate ofCD4＋CD25＋CD127dim/-Tregs cells in peripheral blood of breast cancer patients was statistically significantly higher than that in breast fibroadenoma patients （P〈0. 05 ） ; immunomagnetic cell sorting technique showed that the number of cells in cell counting chamber was 4×10^6 , the positive rate of flow cytometry was 94. 16% , the number of cells was 1×10^6 after five days of in vitro culture, the positive rate of CD4＋CD25＋CD127dim/-Tregs cells detected by flow cytometry was 90.09%. Conclusion： CD4＋CD25＋CD127dim/-Tregscells may be important factors of carcinogenesis and development of breast cancer, Treg cells can be used to discriminate biological behaviors of benign and malignant breast tumors with important reference value; immunomagnetic cell sorting technique, anti-CD3 package, in vitro proliferation and culture by stim- ulating CD28 and IL-2 can obtain CD4＋CD25＋CD127dim/-Tregs cells with ideal purity, which lays a foundation for further studying TCR CDR3 molecular characteristics of Treg cells in peripheral blood of breast cancer patients.