The pulmonary vascular sequestration of neutrophils in endotoxemia is initiated by an effect of endotoxin on the neutrophil in the rabbit

Abstract

Endotoxemia causes neutrophil sequestration in the pulmonary vascular bed. Such sequestration may be a critical initiating event in the generation of microvascular injury, although the mechanisms that lead to this localization are not understood. To investigate these phenomena, the following study employed intravenous pulses of 111Indium-tropolonate-labeled neutrophils (111In-neutrophils), which circulated in the rabbit with normal kinetics and responded in a manner indistinguishable from unlabeled, circulating neutrophils in response to an intravenous injection of purified endotoxic lipopolysaccharide (LPS) or epinephrine. Pulmonary sequestration of 111In-neutrophils was assessed by quantitative external gamma camera scintigraphy of a lung suprahilar region of interest. Noninvasive assessment of radioactivity by this method accurately reflected total lung radioactivity, which was shown by autoradiography to be confined to the injected 111In-neutrophils. Intravenously administered LPS caused a marked, dose-dependent sequestration of 111In-neutrophils in the pulmonary vasculature, and exhaustive ultrastructural autoradiography showed discretely radiolabeled neutrophils located within pulmonary capillaries. A distinct effect was seen with an intravenous injection of as little as 100 ng per rabbit (i.e., 500 pg/ml blood). A 5-min ex vivo pretreatment of 111In-neutrophils with 10 ng to 10 micrograms/ml LPS in heat-inactivated plasma (which resulted in retention of as little as 500 pg LPS per 10(7) neutrophils) also caused dose-dependent pulmonary sequestration of the pretreated 111In-neutrophils but did not cause generalized neutropenia in recipient rabbits. There was no evidence of complement activation on the surface of pretreated neutrophils.(ABSTRACT TRUNCATED AT 250 WORDS)