夹心ELISA检测可溶型LAIR-1方法的建立及其应用的研究

为了探讨血清可溶型白细胞相关免疫球蛋白样受体 1 (sLAIR 1 )的水平与疾病的关系 ,用生物传感器鉴定了 3种抗LAIR 1分子胞膜外区mAb识别的表位 ,并应用 2种识别不同LAIR 1表位的mAb首次建立了检测sLAIR 1的夹心ELISA方法 ,其检测灵敏度达到 1 5 μg/L。以此方法检测了活化的人PBMC培养上清以及正常人和肾综合征出血热 (HFRS )患者血清中sLAIR 1的水平。在经PMA、PHA、LPS和抗CD3mAb活化的人PBMC细胞培养上清中检测到sLAIR 1。 2 4例正常人血清sLAIR 1的平均水平为 (6 2± 3 3) μg/L ,6 2例HFRS患者血清sLAIR 1的平均水平为 (4 7 2± 35 9) μg/L。证明体内和体外存在天然sLAIR 1。HFRS患者血清sLAIR 1水平明显升高。为进一步研究LAIR 1分子的免疫调节功能及其在临床免疫中的应用提供了新的手段

Establishment and Applications of a Sandwich ELISA for Detection of Soluble Leukocyte Associated Immunoglubulin-Like Receptor-1

Sandwich ELISA for detection of soluble leukocyte associated immunoglubulin like receptor 1(sLAIR 1) was established which may be useful investigating the relation of sLAIR 1 with some diseases.The extracellular region epitopes recognized by anti LAIR 1 mAb were identified by Biosensor and two mAb recognizing the distinct epitopes were employed to establish the sandwich ELISA for detection of sLAIR 1 with a detection limit of 1 5 μg/L The sLAIR 1 was detected both in sera from 24 healthy individuals (6 2±3 3)μg/L, and the sera from 62 patients with hemorrhagic fever with renal syndrome(HFRS) (47 2±35 9)μg/L by using this method In addition, sLAIR 1 was also detected in the culture supernants of PBMC stimulated by PMA, PHA, LPS or anti CD3 mAb sLAIR 1 level found in patients with HFRS was higher than those of the healthy individuals This method may provide a new approach to investigate the immuno regulatory role of LAIR 1 in clinical immunity