Contributions of the Mac-1 glycoprotein family to adherence-dependent granulocyte functions: structure-function assessments employing subunit-specific monoclonal antibodies.
Contributions of the Mac-1 glycoprotein family to adherence-dependent granulocyte functions: structure-function assessments employing subunit-specific monoclonal antibodies.
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DOI：
10.4049/jimmunol.137.1.15
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发表时间：
1986-07
影响因子：
4.4
通讯作者：
D. Anderson;L. Miller;F. Schmalstieg;R. Rothlein;T. Springer
中科院分区：
文献类型：
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作者：
D. Anderson;L. Miller;F. Schmalstieg;R. Rothlein;T. Springer
针对Mac-1（α M）、LFA-1（α L）、p150、95（α X）的α亚基或其共同β亚基的MAb用于表征Mac-1糖蛋白家族对粒细胞粘附反应的贡献。这些单克隆抗体在孵育实验中与正常粒细胞的抑制作用表明不同的粘附贡献的每个亚基。在旨在“上调”α M β和α X β复合物表面表达的趋化性条件下，观察到抗α M、α X和β单克隆抗体的粘附显著增加，并抑制粘附。抗β抗体大于抗α M抗体（OKM-10、M1/70、LM 2/1.6和OKM-1），抗α X抗体大于抗α L MAb，显著抑制蛋白质包被玻璃的粘附和白蛋白包被乳胶珠的结合，但抗HLA、AB或抗CR-1 MAb的作用不明显。在粒细胞聚集试验中观察到对C5 a、PMA或f-Met-Leu-Phe的类似抑制等级顺序。显着抑制定向迁移的抗β或抗α M（OKM-1或OKM-10）单克隆抗体中观察到的subagarose，但不是Boyden趋化性测定;抑制依赖于一个连续的细胞暴露于抗Mac-1 α或β在测定过程中，这表明一个连续的新的Mac-1表达所需的定向易位。抗α M单抗（OKM-10大于LM 2/1.6大于M1/70大于OKM-1）或抗α M +抗CR-1单抗组合显著抑制了用C3衍生配体选择性调理的油红-O石蜡或酵母聚糖的吞噬作用，但仅观察到抗β单抗的最小抑制作用，未观察到抗α L或抗α X单抗的作用。类似地，补体依赖性吞噬相关乳铁蛋白释放、摄取和金黄色葡萄球菌502 A的细胞内杀伤以及iC 3b调理SRBC的结合被抗α M抗体（OKM-10，M1/70）或抗α M抗体+抗CR-1单抗的组合显著抑制，但不被抗β、α L或α X单抗抑制。值得注意的是，在粘附非依赖性功能的测定中，抗Mac-1单抗均未表现出抑制作用，这些功能包括形状变化、特异性f-Met-Leu-3 H-Phe结合、O-2生成、化学发光演变或响应PMA的乳铁蛋白释放。（400字处截断摘要）
MAb directed at the alpha-subunits of Mac-1 (alpha M), LFA-1 (alpha L), p150,95 (alpha X), or their common beta-subunit were used to characterize the contributions of the Mac-1 glycoprotein family to granulocyte adherence reactions. Inhibitory effects of these MAb in incubation experiments with normal granulocytes indicated distinct adhesive contributions of each subunit. Significantly greater adherence, and inhibition of adherence by anti alpha M, alpha X, and beta MAb, was observed under chemotactic conditions designed to "up-regulate" the surface expression of the alpha M beta and alpha X beta complexes. Adherence to protein-coated glass and binding of albumin-coated latex beads were significantly inhibited by anti-beta greater than anti-alpha M (OKM-10, M1/70, LM2/1.6 and OKM-1) greater than anti-alpha X greater than anti-alpha L MAb, but no effects of anti-HLA, AB, or anti-CR-1 MAb were evident. A similar rank order of inhibition was observed in granulocyte aggregation assays in response to C5a, PMA, or f-Met-Leu-Phe. Significant inhibition of directed migration by anti-beta or anti-alpha M (OKM-1 or OKM-10) MAb was observed in subagarose but not Boyden chemotaxis assays; inhibition was dependent on a continuous cell exposure to anti-Mac-1 alpha or beta during the assay, suggesting that a continuum of new Mac-1 expression is required for directed translocation. Phagocytosis of Oil-Red-O paraffin or zymosan selectively opsonized with C3-derived ligands was significantly inhibited by anti-alpha M MAb (OKM-10 greater than LM2/1.6 greater than M1/70 greater than OKM-1) or by combinations of anti-alpha M + anti-CR-1 MAb, but only minimal inhibitory effects of anti-beta MAb and no effects of anti-alpha L or anti-alpha X MAb were seen. Similarly, complement-dependent phagocytosis-associated lactoferrin release, ingestion, and intracellular killing of Staphylococcus aureus 502A, and binding of iC3b-opsonized SRBC, were significantly inhibited by anti-alpha M (OKM-10, M1/70) or combinations of anti-alpha M + anti-CR-1 MAb, but not by anti-beta, alpha L, or alpha X MAb. Notably, none of the anti-Mac-1 MAb demonstrated inhibitory effects in assays of adherence-independent functions including shape change, specific f-Met-Leu-3H-Phe binding, O-2 generation, chemiluminescence evolution, or lactoferrin release in response to PMA.(ABSTRACT TRUNCATED AT 400 WORDS)