论文部分内容阅读
目的:研究趋化因子CCL20对小鼠胸腺CD4+CD25+双阳性T细胞发育的影响,为天然调节性T细胞调控的研究提供基础数据。方法:采用14.5d龄胚鼠胸腺进行体外培养,以流式细胞术(FCM)检测不同时间点胸腺CD4+CD25+细胞的变化,同时计数每个胸腺小叶的细胞数变化。结果:体外胸腺培养的第1～6天胸腺细胞比例,细胞数量趋势变化与胸腺体内发育的第14.5、15、16、17、18、19天CD4+CD25+双阳性T细胞的比例,细胞数量的趋势变化相似;在胸腺体外培养的第1～6天,CD4+CD25+T细胞占CD4+T细胞的比例分别为58.29%、12.14%、6.08%、17.78%、9.06%、4.04%,占CD25+T细胞的比例分别为3.75%、10.81%、17.20%、51.93%、61.64%、80.06%,这一发育趋势与体内结果具有一致性。在4mg/L的CCL20干预下,胸腺体外培养的第3、6天CD4+CD25+胸腺细胞分别从3.24±0.18、3.96±0.24下降至1.27±0.11、1.76±0.22(P<0.001)。结论:体外培养的CD4+CD25+双阳性胸腺细胞数量和比例变化与体内发育变化趋势一致,CCL20明显下调胸腺CD4+CD25+的表达,这将为天然调节性T细胞的调控研究提供有效的参考依据。
Objective: To study the effect of chemokine CCL20 on the development of CD4 + CD25 + double positive T cells in thymus of mice and provide the basic data for the study of regulation of natural regulatory T cells. Methods: The thymus of 14.5 d old embryos was cultured in vitro. The changes of CD4 + CD25 + cells in thymus at different time points were detected by flow cytometry (FCM), and the changes of the number of cells in each thymus leaf were counted. RESULTS: The proportion of thymocytes and the number of cells in the first thirteenth day of thymus culture in vitro were correlated with the proportion of CD4 + CD25 + double-positive T cells on the 14th, 15th, 16th, 16th, The percentage of CD4 + CD25 + T cells in CD4 + T cells in vitro were 58.29%, 12.14%, 6.08%, 17.78%, 9.06%, 4.04% + T cells were 3.75%, 10.81%, 17.20%, 51.93%, 61.64%, 80.06%, respectively. The developmental trend was consistent with the in vivo results. The CD4 + CD25 + thymocytes on days 3 and 6 of thymocytes were decreased from 3.24 ± 0.18 and 3.96 ± 0.24 to 1.27 ± 0.11 and 1.76 ± 0.22 (P <0.001) at 4 mg / L CCL20 intervention respectively. CONCLUSION: The change of CD4 + CD25 + double positive thymocytes in vitro is in accordance with the tendency of in vivo development. CCL20 downregulates the expression of CD4 + CD25 +, which will provide an effective reference for the regulation of natural regulatory T cells.