Abstract
In this work, the effects of bacterial LPS, TNF-α, and IFN-γ 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-γ or TNF-α plus IFN-γ became transiently dye coupled (Lucifer yellow) within 24 h. Cells treated with LPS, TNF-α, or IFN-γ alone remained dye uncoupled. In dye-coupled cells, the spread of Lucifer yellow to neighboring cells was reversibly blocked with 18 α-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-α significantly reduced the LPS plus IFN-γ-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-α-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-α plus IFN-γ. 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.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 1320-1328 |
| Number of pages | 9 |
| Journal | Journal of Immunology |
| Volume | 170 |
| Issue number | 3 |
| State | Published - Feb 1 2003 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Immunology
Cite this
TNF-α plus IFN-γ induce connexin43 expression and formation of gap junctions between human monocytes/macrophages that enhance physiological responses. / Eugenin, Eliseo; Brañes, María C.; Berman, Joan W.; Sáez, Juan C.
In: Journal of Immunology, Vol. 170, No. 3, 01.02.2003, p. 1320-1328.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - TNF-α plus IFN-γ induce connexin43 expression and formation of gap junctions between human monocytes/macrophages that enhance physiological responses
AU - Eugenin, Eliseo
AU - Brañes, María C.
AU - Berman, Joan W.
AU - Sáez, Juan C.
PY - 2003/2/1
Y1 - 2003/2/1
N2 - In this work, the effects of bacterial LPS, TNF-α, and IFN-γ 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-γ or TNF-α plus IFN-γ became transiently dye coupled (Lucifer yellow) within 24 h. Cells treated with LPS, TNF-α, or IFN-γ alone remained dye uncoupled. In dye-coupled cells, the spread of Lucifer yellow to neighboring cells was reversibly blocked with 18 α-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-α significantly reduced the LPS plus IFN-γ-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-α-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-α plus IFN-γ. 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.
AB - In this work, the effects of bacterial LPS, TNF-α, and IFN-γ 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-γ or TNF-α plus IFN-γ became transiently dye coupled (Lucifer yellow) within 24 h. Cells treated with LPS, TNF-α, or IFN-γ alone remained dye uncoupled. In dye-coupled cells, the spread of Lucifer yellow to neighboring cells was reversibly blocked with 18 α-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-α significantly reduced the LPS plus IFN-γ-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-α-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-α plus IFN-γ. 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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UR - http://www.scopus.com/inward/citedby.url?scp=0037307922&partnerID=8YFLogxK
M3 - Article
C2 - 12538692
AN - SCOPUS:0037307922
VL - 170
SP - 1320
EP - 1328
JO - Journal of Immunology
JF - Journal of Immunology
SN - 0022-1767
IS - 3
ER -