Klock J C, Stossel T P
J Clin Invest. 1977 Nov;60(5):1183-90. doi: 10.1172/JCI108871.
Granulocytes collected by reversible adhesion to nylon wool fiber (NWF) function relatively well in standard in vitro tests; however, they have an abnormally shortened survival time in the circulation. Assuming that this rapid disappearance represents clearance and that recognition by phagocytes is important for such clearance, we used an autologous in vitro cell:cell recognition assay to determine whether phagocytes can detect cellular changes induced by exposure of normal granulocytes to NWF. Human granulocytes incubated with NWF 1 h at 37 degrees C, eluted with 20% acid citrate dextrose plasma, and washed stimulated the hexose monophosphate shunt activity of normal granulocytes an average of twofold (193+/-40% of controls), indicating a recognition response. NWF-induced granulocyte recognition was not dependent on plasma factors or activated complement components but was dependent on the time that the granulocyte was on the NWF and was maximal by 60 min of exposure. After elution from NWF, granulocytes demonstrated resting glucose oxidation rates only slightly higher than normal; however, during the first 20 min of exposure to NWF, granulocytes increased their rate of (14)CO(2) production from [1-(14)C]glucose three- to five-fold. Therefore, experiments were performed to determine whether toxic oxygen metabolites produced by NWF-adherent cells might contribute to recognition. The results showed that (a) normal granulocytes exposed to NWF in the presence of scavengers of superoxide anion (superoxide dismutase) or free radicals (ascorbate, mannitol, or benzoate) and washed before assay did not stimulate glucose oxidation of indicator granulocytes; and (b) NWF granulocytes prepared from cells unable to generate high levels of toxic oxygen metabolites, i.e. cells prepared anaerobically or from a patient with chronic granulomatous disease, also failed to stimulate indicator granulocytes. Human granulocytes placed in contact with NWF show an oxidative burst and become recognizable to other phagocytes. Free radical scavengers are effective in minimizing this recognition conferred on NWF-procured granulocytes.
通过可逆性黏附于尼龙羊毛纤维(NWF)收集的粒细胞在标准体外试验中功能相对良好;然而,它们在循环中的存活时间异常缩短。假设这种快速消失代表清除,并且吞噬细胞的识别对于这种清除很重要,我们使用了一种自体体外细胞:细胞识别试验来确定吞噬细胞是否能够检测到正常粒细胞暴露于NWF后诱导的细胞变化。将人类粒细胞在37℃下与NWF孵育1小时,用20%酸性枸橼酸盐葡萄糖血浆洗脱并洗涤,刺激正常粒细胞的磷酸己糖旁路活性平均增加两倍(对照组的193±40%),表明存在识别反应。NWF诱导的粒细胞识别不依赖于血浆因子或活化的补体成分,但依赖于粒细胞在NWF上的停留时间,暴露60分钟时达到最大值。从NWF洗脱后,粒细胞的静息葡萄糖氧化率仅略高于正常水平;然而,在暴露于NWF的前20分钟内,粒细胞将[1-(14)C]葡萄糖的(14)CO2产生率提高了三到五倍。因此,进行了实验以确定NWF黏附细胞产生的毒性氧代谢产物是否可能有助于识别。结果表明:(a)在超氧阴离子清除剂(超氧化物歧化酶)或自由基清除剂(抗坏血酸、甘露醇或苯甲酸盐)存在下暴露于NWF并在检测前洗涤的正常粒细胞不会刺激指示粒细胞的葡萄糖氧化;(b)由无法产生高水平毒性氧代谢产物的细胞制备的NWF粒细胞,即厌氧制备的细胞或慢性肉芽肿病患者的细胞,也未能刺激指示粒细胞。与NWF接触的人类粒细胞会出现氧化爆发,并变得可被其他吞噬细胞识别。自由基清除剂可有效减少赋予NWF获取的粒细胞的这种识别。