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OKT3对成熟T细胞有很强的促增殖作用,其对T细胞作用有赖于CD3分子的存在.据报道17周龄后胎儿胸腺细胞(FT)中CD3~+细胞已达60%以上,但关于FT对OKT3的反应性少有报道,本文对此进行了研究,旨在进一步了解FT的功能特性.1 材料和方法1.1 FT和胎儿脾单个核细胞(FSMC)制备 取18～28周龄胎儿胸腺,经200目铜同研磨,Hanks液洗涤后,用含10%小牛血清的1640培液制成FT悬液;胎脾制成单细胞悬液后,经淋巴细胞分离液分出FSMC,洗涤,制成FSMC悬液.1.2 细胞增殖测定 采用~3H-TdRn掺入法.FT和FSMC培养浓度为1×10~6/ml;OKT3(北医大提供小鼠腹水)和IL-2(中科院上海生化所产品)最终浓度分别为10~(-4)稀释和10U/ml;设立OKT3单独和OKT3+IL-2联合刺激两种培养条件,并设相应对照,每组为3个复孔.1.3 免疫荧光技术和流式细胞仪分析 采用间接免疫荧光染色法.抗CD25单抗和FITC标记的羊抗鼠IgG均为DAKO产品.用FACS420进行表型分析.2 结果
OKT3 has a strong pro-proliferative effect on mature T cells and its effect on T cells depends on the presence of CD3 molecules. CD3 + cells in fetal thymocytes (FT) have been reported to have reached more than 60% after 17 weeks of age, but about FT reactivity to OKT3 rarely reported, this article was studied to further understand the functional properties of FT.1 Materials and methods 1.1 FT and fetal spleen mononuclear cells (FSMC) Preparation Take 18 to 28 weeks old fetus thymus , After grinding with 200 mesh copper, Hanks liquid washing, with 10% calf serum 1640 culture solution made of FT suspension; fetal spleen made single cell suspension, the separation of FSMC by lymphocyte separation, washing , Made of FSMC suspension.2.2 Cell proliferation assay using ~ 3H-TdRn incorporation.FT and FSMC culture at a concentration of 1 × 10 ~ 6 / ml; OKT3 (Beiyidai provide mouse ascites) and IL-2 (Chinese Academy of Sciences Shanghai Biochemical products) were diluted to 10 ~ (-4) and 10U / ml, OKT3 alone and OKT3 + IL-2 were used to stimulate the two culture conditions, and the corresponding control, each group of three complex holes .1.3 Indirect immunofluorescence staining was used for immunofluorescence and flow cytometry analysis.Anti-CD25 mAb and FITC-labeled goat anti-mouse IgG were both DAKO products.FACS420 Type analysis .2 results