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CD226单克隆抗体诱导人脐静脉内皮细胞胞质钙离子浓度的变化

陈丽华, 刘雪松, 刘飞, 金伯泉

第四军医大学基础部免疫学教研室，西安 710032

摘要

为观察CD226单克隆抗体(mAb)对培养人脐静脉内皮细胞(HUVECs)胞质钙离子变化的影响，我们用F1 u7o,3作为钙指示剂，用激光共聚焦显微镜观测不同状态下CD226 mAb作用后HUVECs胞质钙离子「Ca ~(2 +)];的变化。结果发现:(1)用Hanks液平衡，CD226 mAb作用后HUVECs [ Cat ` ];水平缓慢升高后回到原位;加入二抗(羊抗鼠IgG)交联后「cat ` ];水平有较大幅度的升高，随后回到原位，与此同时，细胞外液中「 ca~(2+) ` ]o水平有一定程度的下降;(2)用D Hanks液平衡，CD226 mAb作用后HUVECs [ Ca~(2+)` ];水平无显著变化，加入二抗发生交联作用后，[ Ca~(2+) ],水平有较大幅度的下降;(3)用E GTA预处理后，CD226 mAb及其二抗交联对HUVECs [ Ca~(2+) ];变化无显著影响。以上结果提示，CD226 mAb及其二抗交联可诱导不同状态的HUVECs胞质钙离子水平发生不同程度的变化，从而参与一系列的生理和病理过程。

关键词： 生理学; CD226; 激光共聚焦显微镜; [ Ca~(2+) ]; 人脐静脉内皮细胞

CD226 monoclonal antibo街induces variation of intracytosolic free calcium level in human umbilical vein endothelial cells

CHEN Li- Hua, LIU Xue- Song, LIU Fei, JIN Bo- Quan

Department of Immunology, the Fourth Military Unizursity, Xian 710032

Abstract

In order to study the possible mechanism of CD226 monoclonal antibody (mAb)一mediated intracellular message transduction in human umbilical vein endothelial cells(HUVECs)，the influence of CD226 mAl and its cross-linking by secondary antibody(11 Ab) on the concentration changes in[Ca 2十]，in the HUVECs under different conditions were determined妙confocal laser scanning microscopy .The main results are as follows (1)When the culture medium was balanced妙Hanks Buffer，[Ca 2十]，in HUVECs increased slowly after stimu-
lationCD226 mAb，whe re as[ Ca ~(2+)]，increase was accompanied妙[Ca 2十]。decrease after the mAb was crosslinkedgoat anti- mouse IgG.Then[Ca~( 2+)]，and[Ca ~(2+)]。all returned to the normal level.(2) When the culture medium was balanced by Hanks Suffer，[Ca ~(2+)]，in HUVECs showed little variation when the cells wenstimulated CD226 mAb，but[Ca~( 2+)]，decreased markedly after cross-linking.(3) When HUVECs were pretreated with EGTA，there was no variation in[Ca~( 2+)]，of HUVECs after CD226 mAb stimulation alone or cross-
linking of the mAb.Our results suggest that stimulation CD226 mAb and cross-linking by goat anti- mouse IgCinduce the variation of[Ca~( 2+)]，in HLTVECs under different conditions and the variation‘[Ca~( 2+)]，in HLTVEC:
may play an important role in many physiological and pathological processes.

Key words: physiology;CD226;confocal laser scanning microscopy;;

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引用本文：

陈丽华, 刘雪松, 刘飞, 金伯泉. CD226单克隆抗体诱导人脐静脉内皮细胞胞质钙离子浓度的变化[J]. 生理学报 2003; 55 (3): .

CHEN Li- Hua, LIU Xue- Song, LIU Fei, JIN Bo- Quan. CD226 monoclonal antibo街induces variation of intracytosolic free calcium level in human umbilical vein endothelial cells. Acta Physiol Sin 2003; 55 (3): (in Chinese with English abstract).