alexa Regulation of human monocyte apoptosis by the CD14 molecule.


Immunotherapy: Open Access

Author(s): Heidenreich S, Schmidt M, August, Cullen P, Rademaekers A

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Bacterial products such as LPS have been shown to activate monocytes and to increase CD14 expression, while anti-inflammatory cytokines, i.e., IL-4, down-regulate CD14. Furthermore, activation of monocytes increases survival, whereas deactivation evokes apoptosis (programmed cell death, PCD). This correlation among activation, CD14 expression, and the lifespan of the cells prompted us to investigate the role of CD14 in monocyte apoptosis. The effects of LPS and IL-4 on the expression of CD14, indicated by binding of Leu M3 Ab, and PCD of monocytes were studied simultaneously and in a kinetic fashion by multiparameter flow cytometry. Monocyte PCD was determined by binding of FITC-conjugated annexin V, which indicates apoptotic cell death in early stages, and was confirmed using well-established detection methods, i.e., DNA electrophoresis, electron microscopy, or colorimetric DNA staining. The present study shows that the LPS-induced increase in CD14 expression rescued monocytes from apoptosis, whereas IL-4 treatment first down-regulated CD14 expression and consecutively evoked apoptosis. CD14-/annexin V- monocytes were not apoptotic as confirmed by DNA electrophoresis, whereas CD14-/ annexin V+ monocytes showed clear apoptotic features. Kinetic studies ruled out that monocytes first bound annexin V and later lost the CD14 Ag. Other molecules, such as HLA-A, -B, and -C Ags, were not down-regulated during apoptosis. Enzymatic removal of membrane-bound CD14 by phosphatidylinositol-specific phospholipase C evoked PCD similarly to IL-4. These results suggest that regulation of CD14 receptor expression is an early effector mechanism mediating life or death of monocytes. Down-regulation or removal of the receptor triggers apoptosis, whereas up-regulation promotes survival.
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This article was published in J Immunol and referenced in Immunotherapy: Open Access

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