Klock J C, Bainton D F
Blood. 1976 Jul;48(1):149-61.
Granylocyte bactericidal capacity, chemotaxis, hexose monophosphate shung activity (before and after phagocytic stimulus), and quantitative nitroblue tetrazolium reduction and enzyme content were examined in cells obtained by filtration leukaphresis (FL) and continuous-flow centrifugation (CFC). A decrease in the bactericidal efficiency of FL-produced cells compared to that of both normal and CFC-procured granulocytes was found; the decrease was 17% with a cell-to-bacteria ratio of 5:1, and 55% with a 1:1 ratio. Moreover, FL-acquired cells were often vacuolated and consistently contained less acid phosphatase and beta-glucuronidase than did normal granulocytes. When normal cells were incubated for 1-2 hr with nylon wool, 30% of the total acid phosphatase and beta-glucuronidase was released, with no evidence of cell death, thus suggesting degranulation. Similar results were obtained with glass, cotton, or polysulfone plastic fibers. Electron microscopic and peroxidase cytochemical studies of the adherence of normal granulocytes to nylon fibers were also carried out. After 30 min of incubation, cell-to-fiber attachment and cellular aggregation had occurred, although the cells per se appeared normal. After 60 and 120 min, other changes became apparent: (1) a decrease in the amount of cytoplasmic granules; (2) large, intracytoplasmic vaculoles; and (3) extracellular peroxidase on fiber surfaces. We conclude that granulocytes obtained by adherence to nylon fibers show both morphological and biochemical evidence of degranulation and diminished bactericidal capacity, and that these abnormalities may be causally related to decreased granulocyte survival in transfusion recipients.
对通过过滤白细胞分离术(FL)和连续流离心法(CFC)获得的细胞,检测了其粒细胞杀菌能力、趋化性、吞噬刺激前后的磷酸己糖旁路活性、定量硝基蓝四氮唑还原反应及酶含量。结果发现,与正常粒细胞和CFC获得的粒细胞相比,FL产生的细胞杀菌效率降低;细胞与细菌比例为5:1时降低了17%,1:1比例时降低了55%。此外,FL获得的细胞常出现空泡化,且与正常粒细胞相比,其酸性磷酸酶和β-葡萄糖醛酸酶含量始终较低。当正常细胞与尼龙毛一起孵育1 - 2小时后,总酸性磷酸酶和β-葡萄糖醛酸酶的30%被释放,且无细胞死亡迹象,提示脱颗粒。用玻璃、棉花或聚砜塑料纤维也得到了类似结果。还对正常粒细胞与尼龙纤维黏附进行了电子显微镜和过氧化物酶细胞化学研究。孵育30分钟后,细胞与纤维发生附着和细胞聚集,尽管细胞本身看起来正常。60分钟和120分钟后,其他变化变得明显:(1)细胞质颗粒数量减少;(2)出现大的胞质空泡;(3)纤维表面有细胞外过氧化物酶。我们得出结论,通过黏附尼龙纤维获得的粒细胞显示出脱颗粒的形态学和生化证据以及杀菌能力降低,并且这些异常可能与输血受者中粒细胞存活减少存在因果关系。