Neutrophils from patients after burn injury express a deficiency of the oxidase components p47-phox and p67-phox
Neutrophils from patients after burn injury express a deficiency of the oxidase components p47-phox and p67-phox
复制标题
烧伤后患者的中性粒细胞表达氧化酶成分 p47-phox 和 p67-phox 缺乏
DOI：
10.1182/blood.v88.11.4321.bloodjournal88114321
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发表时间：
1996
期刊：
影响因子：
20.3
通讯作者：
Ambruso
中科院分区：
文献类型：
--
作者：
J. Rosenthal;G. Thurman;N. Cusack;V. Peterson;H. Malech;Ambruso
感染是热损伤后患者发病和死亡的主要原因。感染的易感性部分与多形核白细胞（PMN）功能异常和呼吸爆发减弱有关。为探讨严重烧伤后呼吸爆发缺陷的生化基础，对烧伤后呼吸爆发氧化酶系统及其组分的状态进行了研究。从24例12%至62%烧伤患者中分离出中性粒细胞。与健康对照组相比，烧伤后完整的中性粒细胞的氧化酶活性（以超氧阴离子（O2-）产生或耗氧量测量）降低。从患者中性粒细胞的亚细胞组分产生较少的O2-在十二烷基硫酸钠无细胞系统，这是相关的细胞质，但不是质膜的贡献减少。随后，用CM-琼脂糖凝胶分离胞质溶胶，得到两个级分;一个级分含有p47-phox和p67-phox（47/67混合物），另一个级分含有剩余的胞质溶胶组分（运行通过[RT]）。虽然RT对烧伤患者胞浆中氧化酶活性的贡献与对照相似，但烧伤患者PMNs中p47/67混合物的活性不足。使用免疫放射自显影技术的定量分析显示，p47-phox和p67-phox的一致，但显着减少。添加纯化的或人重组p47-phox而不是p67-phox纠正了烧伤患者胞质溶胶中氧化酶活性的降低。因此，减少呼吸爆发活动发现从个人与热损伤的中性粒细胞与一个特定的，定量缺乏p47- phox。
Infection is a major cause of morbidity and mortality in patients after thermal injury. This predisposition to infections is related, in part, to abnormal polymorphonuclear leukocyte (PMN) function and a diminished respiratory burst. To evaluate the biochemical basis for the defective respiratory burst after major burns, the status of the oxidase enzyme system and its components was investigated. PMNs were isolated from 24 patients with 12% to 62% burns. Oxidase activity of intact PMNs, measured as superoxide anion (O2-) generation or oxygen consumption, was decreased in burn compared with healthy controls. Subcellular fractions from patient PMNs generated less O2- in the sodium dodecyl sulfate cell-free system, and this was related to a diminished contribution by cytosol but not by plasma membrane. Subsequently, cytosol was separated with CM-Sepharose, yielding two fractions; one contained the p47-phox and p67-phox (47/67 mix) and the other contained the remaining cytosolic components (run through [RT]). Although the contribution to oxidase activity made by RT from patient cytosol was similar to that of control, the activity of p47/67 mix from PMNs of burn patients was deficient. Quantitative assays using an immunoautoradiographic technique showed a consistent, but significant decrease in both p47-phox and p67-phox. The addition of purified or human recombinant p47-phox but not p67-phox corrected the diminished oxidase activity of cytosol from burn patients. Thus, decreased respiratory burst activity found in PMNs from individuals with thermal injury was associated with a specific, quantitative deficiency of p47- phox.