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Am. J. Respir. Cell Mol. Biol., Volume 23, Number 6, December, 2000 780-787

CD14+ Cells Are Necessary for Increased Survival of Eosinophils in Response to Lipopolysaccharide

JoAnn Meerschaert, William W. Busse, Paul J. Bertics, and Deane F. Mosher

Departments of Medicine and Biomolecular Chemistry, University of Wisconsin, Madison, Wisconsin; and Department of Biological Sciences, St. Cloud State University, St. Cloud, Minnesota

There has been considerable interest in the effect that gram-negative bacterial endotoxin (lipopolysaccharide [LPS]) can have in asthma, given that inhalation of LPS has been shown to cause bronchial hyperresponsiveness. Further, there is evidence that the endotoxin-binding protein CD14 may be a marker for asthma. Inhaled LPS has been shown to cause an influx of eosinophils into the nasal airway and to increase the survival of CD16-negatively selected eosinophils in vitro. In this study, we compared survival of eosinophils isolated via CD16-negative selection with eosinophils that were isolated using both CD16- and CD14-negative selection criteria. Survival of CD16-negatively selected eosinophils was enhanced by LPS in a dose-dependent manner and was inhibited by the endotoxin antagonists polymyxin B or lipid X. In contrast, depletion of CD14+ cells within the eosinophil preparations (CD14/CD16-negatively selected eosinophils) decreased the effect of LPS on survival. Preincubation of CD16-negatively selected eosinophils with antibody 60bd, which blocks LPS binding to CD14, prevented the survival-enhancing effect of LPS. However, CD14 was not detected on eosinophils by flow cytometry, even after incubation with LPS for up to 24 h. These results suggest that the survival-enhancing effect of LPS on eosinophils requires the presence of CD14+ cells in the population. It is our hypothesis that enhanced eosinophil survival with LPS involves the contribution of another cell type.




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