Luscinskas F W, Brock A F, Arnaout M A, Gimbrone M A
Department of Pathology, Brigham and Women's Hospital, Boston, MA 02115.
J Immunol. 1989 Apr 1;142(7):2257-63.
We have examined the contributions of endothelial-leukocyte adhesion molecule-1 (ELAM-1) and the complex of leukocyte surface adhesion molecules designated CD11/CD18 to the adhesion of human polymorphonuclear leukocytes (PMN) to cultured human endothelial cells (HEC), activated by rIL-1 beta for 4 or 24 h. Inhibition of PMN attachment to IL-1-activated HEC was measured in a quantitative in vitro monolayer adhesion assay, after treatment with mAb directed to ELAM-1 (mAb H18/17), and to CD11a (mAb L11), CD11b (mAb 44), CD11c (mAb L29), and CD18 (mAb 10F12), alone or in combination. Pretreatment of activated HEC with mAb H18/7 inhibited PMN adhesion by 47 +/- 8% whereas control mAb had no effect. CD11/CD18-directed mAb significantly blocked PMN adhesion to activated HEC (anti-CD11a, 40 +/- 3%; anti-CD11b, 34 +/- 4%; anti-CD18, 78+/- 6% inhibition). The combination of mAb H18/7 and each of the various anti-CD11/CD18 mAb resulted in greater inhibition of PMN adhesion than any Mab alone. After 24 h of rIL-1 beta treatment, when ELAM-1 was markedly decreased but elevated PMN adhesion was still observed, mAb H18/7 had no effect on PMN adhesion. At this time, CD11/CD18-dependent adhesive mechanisms predominated and a CD11c-dependent mechanism became apparent (anti-CD11a, 67 +/- 4% inhibition; anti-CD11b, 45 +/- 9%; anti-CD11c, 26 +/- 6%; anti-CD18, 97 +/- 1%). In summary, PMN adhesion to IL-1-activated HEC involves both CD11/CD18-dependent mechanisms and an ELAM-1-dependent mechanism, and the relative contribution of these varies at different times of IL-1-induced HEC activation. The additive blocking observed at 4 h with mAb H18/7 in combination with CD11/CD18-directed Mab implies that members of the CD11/CD18 complex do not function as an obligate ligand(s) for ELAM-1.
我们研究了内皮细胞-白细胞黏附分子-1(ELAM-1)以及被称为CD11/CD18的白细胞表面黏附分子复合物,对人多形核白细胞(PMN)黏附于经重组白细胞介素-1β(rIL-1β)激活4小时或24小时的培养人内皮细胞(HEC)的作用。在用针对ELAM-1的单克隆抗体(mAb H18/17)以及针对CD11a(mAb L11)、CD11b(mAb 44)、CD11c(mAb L29)和CD18(mAb 10F12)单独或联合处理后,通过定量体外单层黏附试验测定PMN对IL-1激活的HEC的黏附抑制情况。用mAb H18/7预处理激活的HEC可使PMN黏附减少47±8%,而对照单克隆抗体则无作用。针对CD11/CD18的单克隆抗体显著阻断PMN对激活的HEC的黏附(抗CD11a,40±3%;抗CD11b,34±4%;抗CD18,78±6%抑制)。mAb H18/7与各种抗CD11/CD18单克隆抗体联合使用,对PMN黏附的抑制作用比任何一种单克隆抗体单独使用时都更强。在rIL-1β处理24小时后,此时ELAM-1明显减少,但仍观察到PMN黏附增加,mAb H18/7对PMN黏附无作用。此时,依赖CD11/CD18的黏附机制占主导,且一种依赖CD11c的机制变得明显(抗CD11a,67±4%抑制;抗CD11b,45±9%;抗CD11c,26±6%;抗CD18,97±1%)。总之,PMN对IL-1激活的HEC的黏附涉及依赖CD11/CD18的机制和依赖ELAM-1的机制,且这些机制的相对作用在IL-1诱导的HEC激活的不同时间有所变化。在4小时时观察到mAb H18/7与针对CD11/CD18的单克隆抗体联合使用时有累加阻断作用,这意味着CD11/CD18复合物的成员并非ELAM-1的必需配体。