摘要
为了解尘螨(HDM)抗原对新生儿脐血单个核细胞(CBMC)及成人外周血单个核细胞(PBMC)CD3+ICOS+细胞阳性率、转录因子T-bet、GATA-3、Foxp3 mRNA表达水平以及培养上清中IL-4、IL-10、IFN-γ表达水平的影响。用流式细胞术,检测新生儿CBMC、成人PBMC体外经PHA和/或HDM抗原刺激前、后CD3+ICOS+细胞阳性率;用RT-PCR法检测细胞体外经PHA和/或HDM抗原刺激前、后T-bet、GATA-3以及Foxp3 mRNA表达水平;用ELISA法,检测细胞在体外经PHA和/或HDM刺激前、后培养上清液中IL-4、IL-10、IFN-γ表达水平。结果表明,高剂量HDM抗原显著下调CBMC经PHA刺激后的CD3+ICOS+阳性率(P<0.05),同时也显著上调PHA刺激前其T-bet mRNA表达(P<0.05),而显著下调该类细胞经PHA刺激后的Foxp3 mRNA的表达(P<0.05)。也显著增加其PHA刺激前的IFN-γ分泌(P<0.01),减少其PHA刺激后IL-10分泌(P<0.05)。高剂量HDM抗原对CBMC的作用强于PBMC,可下调CD3+ICOS+阳性率,显著上调PHA刺激前CBMC T-bet mRNA表达,增加PHA刺激前IFN-γ的分泌,减少PHA刺激后IL-10分泌,提示早期接触高剂量HDM抗原对机体免疫功能的影响较强,可促进新生儿Th1样反应,高剂量HDM抗原可能通过作用于下调Foxp3 mRNA的表达而下调CD4+CD25+调节性T细胞的功能。
To investigate the effects of house dust mites on the percentage of CD3^＋ ICOS^＋ cells and the expression levels of transcription factors T-bet, GATA-3 and Foxp3 mRNA and cytokines IL-4, IL-10 and IFN-γ in the culture supernatants of mononucleatex cells in neonatal cord blood and adult peripheral blood, the percentages of CD3^＋ ICOS^＋ cells from neonatal cord blood and adult peripheral blood before and after stimulations with PHA and house dust mite antigen were determined by flow cytometery, the expression levels of T-bet, GATA-3 and Foxp3 mRNA were analysed by RT-PCR assay and the levels of IL- 4, IL-10 and IFN-γ in the culture supernatants were assayed with ELISA. It was demonstrated that the percentage of CD3^＋ ICOS^＋ cells in neonatal cord blood was significantly reduced after stimulation with PHA in comparision with that before stimulation （P〈0.05）, and the levels of T-bet and Foxp3 mRNA in neonatal cord blood were significantly elevated in comaprison with that before stimulation （P〈0. 01）respectively. High dose of house dust mite antigen augumented the secretion of IFN-γ before PHA stimulation （P〈0.01）, but decreasex the secretion of IL-10 after PHA stimulation （P〈0.01）. It is concluded that the effect induced by high dose of house dust mite antigen on mononucleatex cells in neonatal cord blood is stronger than on adult peripheral blood. In addition, this antigen can down-regulate the expression of CD3^＋ ICOS^＋ cells, up-regulate the expression of T-bet mRNA, increase the secretion of IFN-γ before PHA stimulation and decrease the secretion of IL-10 after PHA stimulation, thus inducing a Thl response in neonatal cord blood.
出处
《现代免疫学》
CAS
CSCD
北大核心
2007年第2期151-155,共5页
Current Immunology
基金
重庆市科委计划资助项目(2003-43-97)
