Arnaout M A, Lanier L L, Faller D V
Renal Division, Children's Hospital, Boston, Massachusetts.
J Cell Physiol. 1988 Nov;137(2):305-9. doi: 10.1002/jcp.1041370214.
Adhesion of human monocytes and granulocytes to vascular endothelium plays an important role in migration of these cells to inflammatory sites in tissues. A family of three human leukocyte heterodimeric surface molecules named Mo1, LFA-1, and p150,95 (LeuM5) has been shown to mediate leukocyte adhesion to confluent monolayers of human umbilical vein endothelial cells (HUVE). The relative contribution of each of the three molecules in leukocyte endothelial adhesion was studied using a variety of stimuli. Purified human granulocytes and monocytes were radiolabelled and incubated with HUVE for 45 minutes in a 37 degrees C humidified 5% CO2 incubator in the presence or absence of subunit-specific monoclonal antibodies (MAbs). Adhesion was assessed by quantitation of endothelial cell-associated radioactivity and confirmed by microscopic evaluation. MAbs directed against the alpha subunit of LFA-1 as well as to the beta subunit common to all three antigens significantly inhibited unstimulated monocyte adhesion to HUVE. Small but significant inhibiton was also observed using MAbs directed against Mo1a and p150. Phorbol myristate acetate (PMA)-induced grranulocyte adhesion to HUVE was significantly inhibited by anti-Mo1a and anti-beta, but not by anti-LFA-1a or anti-p150. When HUVE were prestimulated by recombinant IL-1, a different pattern of antigen utilization by granulocytes was observed. MAbs directed against each of the three alpha subunits as well as the common beta subunit all inhibited granulocyte adhesion to HUVE. Furthermore the effect of the three anti-alpha subunit MAbs on granulocyte-HUVE adhesion was additive. These studies show that relative contribution of Mo1, LFA-1, and p150,95 to leukocyte endothelial adhesion varies depending on the cell type and the stimulus used. These studies also reveal a novel role for p150,95 in promoting monocyte and granulocyte adhesion to HUVE.
人单核细胞和粒细胞与血管内皮的黏附在这些细胞迁移至组织炎症部位的过程中起着重要作用。已证明一个由三种人白细胞异二聚体表面分子组成的家族,即Mo1、淋巴细胞功能相关抗原-1(LFA-1)和p150,95(LeuM5),可介导白细胞与人脐静脉内皮细胞(HUVE)融合单层的黏附。使用多种刺激物研究了这三种分子中每种分子在白细胞与内皮黏附中的相对作用。纯化的人粒细胞和单核细胞用放射性标记,并在37℃、5%二氧化碳饱和湿度的培养箱中,在有或无亚单位特异性单克隆抗体(MAb)的情况下与HUVE孵育45分钟。通过定量内皮细胞相关放射性评估黏附情况,并通过显微镜评估进行确认。针对LFA-1α亚单位以及所有三种抗原共有的β亚单位的单克隆抗体显著抑制未刺激的单核细胞与HUVE的黏附。使用针对Mo1a和p150的单克隆抗体也观察到了轻微但显著的抑制作用。佛波酯肉豆蔻酸酯乙酸酯(PMA)诱导的粒细胞与HUVE的黏附被抗Mo1a和抗β单克隆抗体显著抑制,但抗LFA-1a或抗p150单克隆抗体则无此作用。当HUVE用重组白细胞介素-1预刺激时,观察到粒细胞利用抗原的不同模式。针对三种α亚单位以及共同的β亚单位的单克隆抗体均抑制粒细胞与HUVE的黏附。此外,三种抗α亚单位单克隆抗体对粒细胞-HUVE黏附的作用是相加的。这些研究表明,Mo1、LFA-1和p150,95对白细胞与内皮黏附的相对作用因细胞类型和所用刺激物而异。这些研究还揭示了p150,95在促进单核细胞和粒细胞与HUVE黏附中的新作用。