摘要
目的体外建立可长期培养的抗原特异性的T辅助细胞(Th)1/Th2克隆及细胞系,并分析其对抗原的反应情况及其产生细胞因子的情况。方法采用卵清白蛋白(OVA)免疫BALB/c小鼠,取淋巴结细胞在体外用同系脾细胞和OVA反复刺激,然后分别用白介素(IL)-2、γ干扰素(IFN-γ)及IL-2I、L-4共同培养,并同时加入同系脾细胞和OVA,进行Th1和Th2克隆的诱导,然后通过有限稀释法进行克隆化培养。对克隆化的细胞用酶联免疫吸附试验(ELISA)和流式细胞仪分别检测其分泌细胞因子的情况及细胞的表面标志。结果各获得2株Th1/Th2细胞系,它们分泌的细胞因子分别为IL-2、IFN-γ和IL-4I、L-5I、L-10等。结论成功建立抗原特异性Th1/Th2细胞系。
Objective To construct a long-term cultured antigen-specific T helper 1 and T helper 2 clone and cell line, and analyze the cytokines secreted. Methods Lymph node T cells from BALB/c mice immunized with OVA were repeatedly stimulated with OVA and splenic cells from BALB/e mice to construct the OVA-spe cific T cell line in vitro. After that,T cells were stimulated with IL-2 andγ-IFN（Thl skewing condition） or IL-2 and IL-4（Th2 skewing condition） for 10 to 14 days. ELISA and flow cytometry were used to measure the cytokines secreted by these cells and their surface markers. Results We obtained two cell lines of each subset,which secreted IL-2 and γ-IFN or IL-4 and IL-5 respectively. Conclusions The antigen-speeipic Th1/Th2 cell line is sucssesfully constructed for future use. （Shanghai Med J, 2005, 28 ： 1000-1003）
出处
《上海医学》
CAS
CSCD
北大核心
2005年第12期1000-1003,共4页
Shanghai Medical Journal
