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目的探讨新生儿出生时淋巴细胞功能低下的可能环节。方法给予成人外周血和脐血淋巴细胞共刺激(抗CD3、抗CD28单抗)和跨膜刺激(phorbolmyristateacetate,PMA和ionomycin,IM)后,应用犤3H犦-TdR掺入法检测淋巴细胞增殖反应;WesternBlot法检测淋巴细胞磷脂酶Cγ1(phospholipaseCγ1,PLCγ1)的表达。结果抗CD3+抗CD28单抗共刺激后,脐血淋巴细胞的增殖反应明显增强;PMA+IM跨膜刺激后,脐血淋巴细胞增殖反应的改善更接近于成人。给予抗CD3+抗CD28单抗共刺激后,脐血淋巴细胞PLCγ1的表达低于成人外周血;刺激前后脐血淋巴细胞PLCγ1表达差异无显著性,而成人外周血淋巴细胞PLCγ1表达则有显著增加。结论脐血淋巴细胞活化的下游途径可以发挥更接近于成人的作用,推测其功能障碍可能主要在于淋巴细胞活化的早期阶段。
Objective To explore the possible link of neonatal lymphocyte dysfunction at birth. Methods After being stimulated with anti-CD3, anti-CD28 and anti-CD28 monoclonal antibodies (PBMCs) by adult peripheral blood and umbilical cord blood lymphocytes, the proliferation of lymphocytes was detected by 3H-TdR incorporation assay The expression of phospholipase Cγ1 (PLCγ1) in lymphocytes was detected by Western blot. Results The co-stimulation with anti-CD3 + anti-CD28 mAb significantly enhanced the proliferation of cord blood lymphocytes. After transmembrane stimulation with PMA + IM, the proliferation of cord blood lymphocytes was more similar to that of adult. After co-stimulated with anti-CD3 + anti-CD28 monoclonal antibody, the expression of PLCγ1 in umbilical cord blood lymphocytes was lower than that in adult peripheral blood. The expression of PLCγ1 in umbilical cord blood lymphocytes was not significantly different before and after stimulation, while the expression of PLCγ1 in adult peripheral blood lymphocytes was significantly increased. Conclusion Downstream pathway of cord blood lymphocyte activation can play a more role in adults, suggesting that dysfunction may mainly lie in the early stage of lymphocyte activation.