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The Journal of Immunology, 2003, 170: 1320-1328.
Copyright © 2003 by The American Association of Immunologists

TNF-{alpha} Plus IFN-{gamma} Induce Connexin43 Expression and Formation of Gap Junctions Between Human Monocytes/Macrophages That Enhance Physiological Responses 1

Eliseo A. Eugenín2,*,{dagger}, María C. Brañes*, Joan W. Berman{dagger} and Juan C. Sáez*,{ddagger}

* Departamento de Ciencias Fisiológicas, Pontificia Universidad Católica de Chile, Santiago, Chile; and Departments of {dagger} Pathology and {ddagger} Neuroscience, Albert Einstein College of Medicine, Bronx, NY 10461

In this work, the effects of bacterial LPS, TNF-{alpha}, and IFN-{gamma} on gap junctional communication (dye coupling) and on the expression of connexin43 (immunofluorescence, immunoblotting, and RT-PCR) in monocytes/macrophages were studied. Freshly isolated human monocytes plated at high density and treated either with LPS plus IFN-{gamma} or TNF-{alpha} plus IFN-{gamma} became transiently dye coupled (Lucifer yellow) within 24 h. Cells treated with LPS, TNF-{alpha}, or IFN-{gamma} alone remained dye uncoupled. In dye-coupled cells, the spread of Lucifer yellow to neighboring cells was reversibly blocked with 18 {alpha}-glycyrrhetinic acid, a gap junction blocker, but it was unaffected by oxidized ATP or probenecid, which block ionotropic ATP-activated channels and organic anion transporters, respectively. Abs against TNF-{alpha} significantly reduced the LPS plus IFN-{gamma}-induced increase in dye coupling. In dye-coupled monocytes/macrophages, but not in control cells, both connexin43 protein and mRNA were detected, and their levels were higher in cells with an elevated incidence of dye coupling. In dye-coupled cells, the localization of connexin43 immunoreactivity was diffuse at perinuclear regions and thin cell processes. The addition of 18-{alpha}-glycyrrhetinic acid induced a profound reduction of monocyte/macrophage transmigration across a blood brain barrier model. It also induced a significant reduction in the secretion of metalloproteinase-2 in cells treated with TNF-{alpha} plus IFN-{gamma}. We propose that some monocyte/macrophage responses are coordinated by connexin-formed membrane channels expressed transiently at inflammatory sites in which these cells form aggregates.




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