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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 investigate the expression and significance of CD4 + CD25 + CD127dim / - regulatory T cells (Treg) in peripheral blood of patients with breast cancer and negative control (fibromatosis) patients. To explore the use of magnetic beads sorting technique to classify human (breast cancer patients ) Peripheral blood CD4 + CD25 + CD127dim / -Treg cells and in vitro culture methods. Methods Flow cytometry was used to detect the expression of CD4 + CD25 + CD127adim / -Treg in peripheral blood of 15 patients with simple breast cancer and 15 patients with simple fibroadenoma. The correlations were analyzed. Immunomagnetic sorting CD4 + CD25 + CD127dim / -Treg cells in peripheral blood of patients with cancer; anti-CD3 coating, CD28, IL-2 stimulation of proliferation in vitro, purity was detected by flow cytometry. Results The positive rate of CD4 + CD25 + CD127dim / -Treg in peripheral blood of breast cancer patients was significantly higher than that of breast fibroids (P <0.05). The peripheral blood CD4 + CD25 + CD127dim / -Tregs The number of plate-counted cells was 4 × 105, the positive rate of flow cytometry was 94.16%, and the number of proliferating cells was 1 × 106 after 5 days. The positive rate of flow cytometry for CD4 + CD25 + CD127dim / -Treg was 90.09 %. Conclusion CD4 + CD25 + CD127dim / -Tregs cells may be one of the important factors in the development of breast cancer. Regulatory T cells can be used as a marker for distinguishing the biological behavior of benign and malignant breast tumors. Immunomagnetic bead sorting CD40, CD28 and IL-2 stimulated the proliferation of CD4 + CD25 + CD127dim / -Tregs cells in vitro, which will lay a foundation for the further study on the molecular characteristics of TCR CDR3 in peripheral blood Tregs of patients with breast cancer .