Iwabuchi K, Yamashita T
Department of Biochemistry, School of Medicine, Juntendo University, Tokyo, Japan.
Blood. 1990 Dec 1;76(11):2368-73.
The effect of constituents of human platelets on leukocyte adherence was examined. Adherence-inhibiting factors (AIFs), which strongly inhibited neutrophil adherence to glass, were present in both cytosol and granule fractions of human platelets. On the Superose 6 gel chromatography (Pharmacia LKB Biotechnology, Uppsala, Sweden), the granular AIF was eluted as a single active peak (2,600 Kd), whereas cytosolic AIFs were eluted at two different positions (2,600 and 480 Kd). When platelets were stimulated by thrombin, granular AIF was released extracellularly without releasing a cytosolic marker. Using DE32 anion exchange chromatography and Superose 6 gel filtration, granular AIF was completely purified. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis suggests that granular AIF consists of two subunits with molecular masses of approximately 340 and 190 Kd. Purified granular AIF inhibited human neutrophil adherence to glass, plastic, and type IV collagen-coated plastic, whereas it did not affect monocyte adherence. These results suggest that granular AIF inhibits neutrophil adherence not only via nonspecific adsorption sites, but also via type IV collagen receptors.
研究了人血小板成分对白细胞黏附的影响。人血小板的胞质溶胶和颗粒组分中均存在强烈抑制中性粒细胞黏附于玻璃的黏附抑制因子(AIFs)。在Superose 6凝胶色谱法(瑞典乌普萨拉的Pharmacia LKB生物技术公司)中,颗粒状AIF以单一活性峰(2600 Kd)洗脱,而胞质溶胶AIFs在两个不同位置(2600和480 Kd)洗脱。当血小板受到凝血酶刺激时,颗粒状AIF释放到细胞外,而未释放胞质溶胶标志物。通过DE32阴离子交换色谱法和Superose 6凝胶过滤,颗粒状AIF被完全纯化。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析表明,颗粒状AIF由两个亚基组成,分子量分别约为340和190 Kd。纯化的颗粒状AIF抑制人中性粒细胞黏附于玻璃、塑料和IV型胶原包被的塑料,而不影响单核细胞黏附。这些结果表明,颗粒状AIF不仅通过非特异性吸附位点,而且通过IV型胶原受体抑制中性粒细胞黏附。