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American Journal of Pathology, Vol 120, 79-86, Copyright © 1985 by American Society for Investigative Pathology

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Uptake and subcellular localization of bacterial lipopolysaccharide in the adrenal gland

JC Mathison and RJ Ulevitch

For determination of the kinetics of uptake and subcellular localization of lipopolysaccharide (LPS) from LPS-high density lipoprotein (LPS-HDL) complexes in the adrenal gland, LPS-HDL complexes were isolated by immunoaffinity chromatography of 125I-Salmonella minnesota Re595 LPS that had been incubated with 20 mM EDTA-rabbit plasma. After intravenous injection of LPS-HDL complexes in rabbits, preferential uptake of the LPS was observed in the adrenal, so that by 5 hours, adrenal-tissue-bound LPS concentrations (determined by use of 131I-BSA blood marker) exceeded all other tissues examined, including liver and spleen, by at least three-fold. For determination of the subcellular localization of LPS, cholesterol-rich (lipid droplet) fractions and cholesterol-depleted fractions were obtained by ultracentrifugation of homogenates of adrenal tissue from rabbits killed at various times after injection of LPS-HDL complexes. As much as 40% of the adrenal-tissue-bound LPS was recovered in the cholesterol- rich fraction 2.5-24 hours after injection of LPS-HDL complexes. Electron-microscopic autoradiographic and immunocytochemical analysis of adrenal cortex of animals killed 5 hours after injection of LPS-HDL complexes demonstrated specific localization of LPS in lipid droplets. These data thus provide direct evidence for the uptake of LPS into the adrenal cortex of animals with intravascular LPS-HDL complexes and indicate that further study of the effect of LPS on adrenocortical function is warranted.

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