摘要
目的 为深入研究T细胞功能 ,探讨一种更为实用可靠的实验方法。方法 应用白细胞表面分化抗原 2 8(CD2 8)单克隆抗体 (单抗 )共刺激检测T细胞功能 ,并观察其影响因素。CD2 8单抗协同CD3单抗或植物血凝素 (PHA)诱导外周血淋巴细胞增殖转化及细胞因子 (白细胞介素 2、4及γ干扰素 )产生 ,分别用氚胸腺嘧啶 (3 H TdR)掺入法和酶联免疫法检测。结果 单独CD2 8单抗几乎不能诱导外周血淋巴细胞活化及细胞因子产生 ;CD2 8协同CD3或PHA刺激外周血淋巴细胞增殖效应的增殖指数 (GI)分别为 2 0 .1± 9.5及 5 3.9± 17.2 ,与单用CD3或PHA (G .I为 10 .6± 4.9及 2 2 .0±7.2 )比较 ,差异有非常显著意义 ;CD2 8共刺激显著促进细胞因子产生 ,与CD3或PHA诱生比较约提高 0 .7～ 10倍 ;由CD2 8单抗共刺激诱导的外周血淋巴细胞转化能力受其浓度和培养时间影响 ;CD2 8单抗共刺激所需浓度在 0 .1～ 10mg/L对外周血淋巴细胞均有活化作用。结论 CD2 8单抗共刺激可作为检测T细胞功能的有效方法。
Objective To find a reliable mtthod for detecting T cell′s functions induced by anti CD28 mAb costimulated.Methods The CD28 mAb costimulated with CD3 mAb or non characteristic mitogen PHA induced T lymphocyte transformation and cytokines (interleukin 2, 4 and interferon γ) production by means of the 3H TdR incorporation, bioassay, and ELISA method. Results Anti CD28 mAb did not induce PBL proliferation and cytokines production alone. The G.I of CD28 mAb costimulated with CD3 mAb or PHA (20.1±9.5,53.9±17.2) stimulating PBL proliferation were more significantly increased than that of CD3 mAb or PHA (10.6±4.9,22.0±7.2) used alone( P <0.05, P <0.001). CD28 mAb costimulated with CD3 mAb or PHA dramatically increased cytokine production for about 0.7～10 times, as compared to CD3mAb or PHA stimulation alone. The PBL activation stimulated with CD28 mAb costimulation was influenced by culture time and the concentration of CD28 mAb. The concentration of CD28 mAb costimulation in 0.1～10 μg/ml induced PBL proliferation.Conclusion The CD28 mAb costimulation may be useful for detecting T cells functions.
出处
《中华检验医学杂志》
CAS
CSCD
2000年第4期226-228,共3页
Chinese Journal of Laboratory Medicine
关键词
共刺激检测
CD28单克隆抗体
T细胞
T lymphocytes
Antigens, differentiation
Antibodies, monoclonal
