SUBCELLULAR-LOCALIZATION OF CD66, CD67, AND NCA IN HUMAN NEUTROPHILS
SUBCELLULAR-LOCALIZATION OF CD66, CD67, AND NCA IN HUMAN NEUTROPHILS
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DOI：
10.1002/jlb.52.1.11
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发表时间：
1992-07-01
影响因子：
5.5
通讯作者：
SKUBITZ, KM
中科院分区：
文献类型：
--
作者：
DUCKER, TP;SKUBITZ, KM
CD 66和CD 67是粒细胞特异性活化抗原;当中性粒细胞活化时，其表面表达上调。CD 66抗体识别一种约180 kd的中性粒细胞表面蛋白，该蛋白也被抗癌胚抗原（CEA）抗体识别，因此是一种非特异性交叉反应抗原（NCA）。CD 67抗体识别通过糖基-磷脂酰肌醇锚附着于膜的约100 kd的中性粒细胞表面蛋白。为了鉴定CD 66和CD 67可以上调的细胞内库，研究了由CD 66和CD 67单克隆抗体和多克隆抗CEA识别的蛋白质的亚细胞分布。通过氮气空化和差速离心制备中性粒细胞质膜、颗粒和细胞质，然后通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和免疫印迹进行分析。大多数的180-kd的蛋白质识别的CD 66抗体和100-kd的蛋白质识别的CD 67抗体位于次级颗粒部分，与质膜部分中检测到的量较少。几个NCA物种范围从约40至200 kd的确定，这些NCA的分布是不同的初级颗粒，次级颗粒，和质膜组分。质膜部分中的主要NCA为约95至100和约180至200 kd;次级颗粒部分含有约42、85、95至100和180至200 kd的主要NCA。在初级颗粒部分中也检测到NCA，最显著的是约90-100 kd;在初级颗粒中未检测到约180 - 200 kd的NCA。次级颗粒中存在CD 66、CD 67和NCA，表明次级颗粒可能是这些抗原被募集到细胞表面并活化的来源。NCA在初级颗粒中的潜在作用尚不清楚。
CD66 and CD67 are granulocyte-specific activation antigens; their surface expression is up-regulated when neutrophils are activated. CD66 antibodies recognize an approximately 180-kd neutrophil surface protein that is also recognized by anti-carcinoembryonic antigen (CEA) antibodies and is therefore a nonspecific cross-reacting antigen (NCA). CD67 antibodies recognize an approximately 100-kd neutrophil surface protein that is attached to the membrane via a glycosyl-phosphatidylinositol anchor. To identify an intracellular pool from which CD66 and CD67 could be up-regulated, the subcellular distribution of proteins recognized by CD66 and CD67 monoclonal antibodies and polyclonal anti-CEA was studied. Neutrophil plasma membranes, granules, and cytoplasm were prepared by nitrogen cavitation and differential centrifugation and then analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting. Most of the 180-kd protein recognized by CD66 antibodies and the 100-kd protein recognized by CD67 antibodies were located in the secondary granule fraction, with lesser amounts detectable in the plasma membrane fraction. Several NCA species ranging from approximately 40 to 200 kd were identified, and the distribution of these NCAs was different in the primary granules, secondary granules, and plasma membrane fractions. The major NCAs in the plasma membrane fraction were of approximately 95 to 100 and approximately 180 to 200 kd; the secondary granule fraction contained major NCAs of approximately 42, 85, 95 to 100, and 180 to 200 kd. NCAs were also detected in the primary granule fraction, the most prominent being of approximately 90-100 kd; no NCA of approximately 180 to 200 kd was detected in the primary granules. The presence of CD66, CD67, and NCAs in the secondary granules suggests secondary granules as a likely source from which these antigens could be recruited to the cell surface with activation. The potential role for NCAs in the primary granules is unknown.