Mayer A M, Zhang P, Spitzer J A
Department of Physiology, Louisiana State University Medical Center, New Orleans 70112-1393, USA.
Shock. 1994 Oct;2(4):289-95. doi: 10.1097/00024382-199410000-00010.
The purpose of this investigation was to determine the effect of anti-rat CD11b/c monoclonal antibody (MAb) on in vitro superoxide anion (O2-) generation in in vivo Escherichia coli lipopolysaccharide (LPS) and tumor necrosis-alpha (TNF)-treated rat polymorphonuclear leukocytes (PMN). After a continuous infusion of a nonlethal dose of E. coli LPS (.5 mg/kg) or TNF (6.0 x 10(5) units) into rats, PMN were recovered by centrifugal elutriation and discontinuous density gradient centrifugation from the liver (LPS-treated) and whole blood (LPS- and TNF-treated) and compared for CD11b/c, CD11a, and CD18 upregulation and their capacity for basal and agonist-stimulated O2- production. Immunofluorescence flow cytometry studies of rat whole blood PMN demonstrated that, upon LPS infusion, there was a time-dependent upregulation of CD11b/c and CD18, but not of CD11a. Similarly, TNF infusion upregulated CD11b/c although to a lesser degree. Stimulation of LPS- and TNF-treated PMN with phorbol 12-myristate 13-acetate (PMA), opsonized zymosan (OPZ), and anti-rat CD11b/c MAb triggered O2- generation. Although total O2- generated by OPZ and anti-rat CD11b/c MAb was less than that generated by PMA stimulation, the in vivo LPS- and TNF-induced beta 2 integrin upregulation did not result in a statistically significant enhancement of O2- generation with respect to normal saline-treated PMN. Our results do not appear to support the hypothesis that enhanced expression of CD11b/c or CD18 might be associated with enhanced in vitro anti-CD11b/c MAb-triggered O2- generation in LPS- and TNF-treated PMN in vivo.
本研究的目的是确定抗大鼠CD11b/c单克隆抗体（MAb）对体内经大肠杆菌脂多糖（LPS）和肿瘤坏死因子-α（TNF）处理的大鼠多形核白细胞（PMN）体外超氧阴离子（O2-）生成的影响。向大鼠持续输注非致死剂量的大肠杆菌LPS（0.5mg/kg）或TNF（6.0×10^5单位）后，通过离心淘洗和不连续密度梯度离心从肝脏（LPS处理组）和全血（LPS和TNF处理组）中回收PMN，并比较其CD11b/c、CD11a和CD18的上调情况以及基础和激动剂刺激下O2-产生的能力。大鼠全血PMN的免疫荧光流式细胞术研究表明，输注LPS后，CD11b/c和CD18呈时间依赖性上调，而CD11a未上调。同样，输注TNF也上调了CD11b/c，尽管程度较小。用佛波醇12-肉豆蔻酸酯13-乙酸酯（PMA）、调理酵母聚糖（OPZ）和抗大鼠CD11b/c MAb刺激LPS和TNF处理的PMN可触发O2-生成。尽管OPZ和抗大鼠CD11b/c MAb产生的总O2-量少于PMA刺激产生的量，但体内LPS和TNF诱导的β2整合素上调并未导致与生理盐水处理的PMN相比O2-生成有统计学意义的增强。我们的结果似乎不支持以下假设：在体内LPS和TNF处理的PMN中，CD11b/c或CD18表达增强可能与体外抗CD11b/c MAb触发的O2-生成增强有关。
