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Luminescence studies of the phagocyte response to endotoxin infusion into normal human subjects: multiple discriminant analysis of luminescence response and correlation with phagocyte morphologic changes and release of elastase

Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, USA, Marie-Brewer{at}omrf.ouhsc.edu

P.A. Haddad

University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, USA

Gary Kinasewitz

University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, USA

Alvin Chang

Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, USA

Glenn Peer

Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, USA

Robert C. Allen

Emory University School of Medicine, Atlanta, Georgia, USA

A blood luminescence system (BLS) was employed to analyze blood phagocyte function in response to infusion of endotoxin (4 ng Escherichia coli lipopolysaccharide (LPS)/kg body weight) into 7 healthy human subjects. The subjects were closely monitored clinically, and extensive chemical, hematological and coagulation measurements were taken during the pretreatment, early (symptomatic, 1—8 h post-LPS), and late (asymptomatic, 12—48 h post-LPS) phases of acute inflammation. BLS assessment included measurement of basal and PMA-stimulated phagocyte oxidase and myeloperoxidase (MPO) activities, and also included measurement of circulating (COR) and PAF-primed maximum (MOR) opsonin receptor-dependent phagocytic activities. Basal oxidase activity peaked at T+1 h and showed an additional peak at T+24 h post-LPS. The COR activity also peaked at 1—2 h, but remained elevated through T+24 h post-LPS, while the basal MPO activity peaked only once at T+1 h. We concluded that while MPO evidence of phagocyte respiratory activation returned to baseline by T+ 4 h, COR evidence of receptor expression (receptor alert) remained elevated through T+24 h. During this early (0—8 h) period, elastase/ 1AT complex concentration peaked at T+3—4 h and again at T+8 h. Peak numbers of circulating polarized and vacuolated phagocytes also appeared at T+3 h and 7 h. We concluded that there was biochemical and morphological evidence of continuing phagocyte activity beyond T+4 h to T+8 h, and that this corresponded with the period during which the subjects were symptomatic. In addition, the appearance of a second peak of basal oxidase activity at T+24 h, multiple discriminant analyses of all the luminescence data, and the sustained elevation of lactate suggested that there was a later second stage (T+12 h to 48 h) of the human response to endotoxin, during which time the subjects were asymptomatic.

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