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目的探讨脐血T细胞中活化T细胞核因子(NF AT)的活性与IL 4基因表达调控的关系。方法采用ELISA和RT PCR法测定受抗CD3 +PMA刺激后 ,18例脐血单个核细胞(CBMC)IL 4基因及其mRNA的表达 ;电泳迁移率转换试验(EMSA) ,测定10例CBMCNF AT的活性。22例成人外周血单个核细胞(PBMC)为对照。结果CBMC培养上清液中IL 4为(12.4±1.5)ng/L ,较PBMC明显低下〔(37.5±5.5)ng/L ,P∨0.01〕 ;CBMCIL 4mR NA的表达较PBMC明显减少 ;PBMC核蛋白内NF AT的活性较强 ,CBMC则几乎测不出NF AT的活性。结论在转录水平缺乏NF AT调控与脐血T细胞IL 4mRNA的转录水平低下有关 ;也是脐血T细胞表达IL 4水平低下的重要原因。
Objective To investigate the relationship between the activity of nuclear factor of activated T cell (NF AT) and the regulation of IL 4 gene in cord blood T cells. Methods The expressions of IL 4 gene and its mRNA in 18 cases of cord blood mononuclear cells (CBMC) stimulated by anti-CD3 + PMA were measured by ELISA and RT-PCR. Electrophoretic mobility shift assay (EMSA) was used to detect the expression of IL-4 in 10 cases of CBMCNF AT active. Twenty-two adult peripheral blood mononuclear cells (PBMCs) served as controls. Results IL-4 in CBMC culture supernatant was (12.4 ± 1.5) ng / L, which was significantly lower than that in PBMC [(37.5 ± 5.5) ng / L, P ∨ 0.01〕; CBMCIL 4mRNA expression was significantly lower than PBMC NF AT protein activity is strong, CBMC almost can not detect NF AT activity. Conclusions The lack of regulation of NF AT at transcriptional level is related to the low transcription of IL 4 mRNA in cord blood T cells. It is also an important reason for the low expression of IL 4 in cord blood T cells.