Pino-Otín M R, Viñas O, de la Fuente M A, Juan M, Font J, Torradeflot M, Pallarés L, Lozano F, Alberola-Ila J, Martorell J
Servei d'Immunologia, Hospital Clinic i Provincial, Barcelona, Spain.
J Immunol. 1995 Mar 15;154(6):3015-24.
CD50 (ICAM-3) is a leukocyte differentiation Ag expressed almost exclusively on hemopoietic cells, with a key role in the first steps of immune response. To develop a specific sandwich ELISA to detect a soluble CD50 form (sCD50), two different mAbs (140-11 and 101-1D2) recognizing non-overlapping epitopes were used. sCD50 was detected in the supernatant of stimulated PBMCs, with the highest levels after CD3 triggering. Simultaneously, the CD50 surface expression diminished during the first 24 h. sCD50 isolated from culture supernatant and analyzed by immunoblotting showed an apparent m.w. of 95 kDa, slightly smaller than the membrane form. These data, together with Northern blot kinetics analysis, suggest that sCD50 is cleaved from cell membrane. Furthermore, we detect sCD50 in normal human sera and higher levels in sera of systemic lupus erythematosus (SLE) patients, especially in those in active phase. The sCD50 levels showed a positive correlation with sCD27 levels (r = 0.4213; p = 0.0026). Detection of sCD50, both after in vitro CD3 triggering of PBMCs and increased in SLE sera, suggests that sCD50 could be used as a marker of lymphocyte stimulation.
CD50(细胞间黏附分子-3)是一种几乎仅在造血细胞上表达的白细胞分化抗原,在免疫反应的起始步骤中起关键作用。为开发一种特异性夹心酶联免疫吸附测定法(ELISA)来检测可溶性CD50形式(sCD50),使用了两种识别不重叠表位的不同单克隆抗体(140-11和101-1D2)。在受刺激的外周血单个核细胞(PBMC)的上清液中检测到sCD50,在CD3触发后水平最高。同时,CD50的表面表达在最初24小时内减少。从培养上清液中分离并通过免疫印迹分析的sCD50显示表观分子量为95 kDa,略小于膜形式。这些数据,连同Northern印迹动力学分析,表明sCD50是从细胞膜上裂解下来的。此外,我们在正常人血清中检测到sCD50,而在系统性红斑狼疮(SLE)患者的血清中水平更高,尤其是在活动期患者中。sCD50水平与sCD27水平呈正相关(r = 0.4213;p = 0.0026)。在体外PBMC经CD3触发后以及SLE血清中sCD50水平升高,这表明sCD50可作为淋巴细胞刺激的标志物。