A role for connexin43 in macrophage phagocytosis and host survival after bacterial peritoneal infection.
The pathways that lead to the internalization of pathogens via phagocytosis remain incompletely understood. We now demonstrate a previously unrecognized role for the gap junction protein connexin43 (Cx43) in the regulation of phagocytosis by macrophages and in the host response to bacterial infection of the peritoneal cavity. Primary and cultured macrophages were found to express Cx43, which localized to the phagosome upon the internalization of IgG-opsonized particles. The inhibition of Cx43 using small interfering RNA or by obtaining macrophages from Cx43 heterozygous or knockout mice resulted in significantly impaired phagocytosis, while transfection of Cx43 into Fc-receptor expressing HeLa cells, which do not express endogenous Cx43, conferred the ability of these cells to undergo phagocytosis. Infection of macrophages with adenoviruses expressing wild-type Cx43 restored phagocytic ability in macrophages from Cx43 heterozygous or deficient mice, while infection with viruses that expressed mutant Cx43 had no effect. In understanding the mechanisms involved, Cx43 was required for RhoA-dependent actin cup formation under adherent particles, and transfection with constitutively active RhoA restored a phagocytic phenotype after Cx43 inactivation. Remarkably, mortality was significantly increased in a mouse model of bacterial peritonitis after Cx43 inhibition and in Cx43 heterozygous mice compared with untreated and wild-type counterparts. These findings reveal a novel role for Cx43 in the regulation of phagocytosis and rearrangement of the F-actin cytoskeleton, and they implicate Cx43 in the regulation of the host response to microbial infection.
Anand
Rahul J
RJ
Division of Pediatric Surgery, Children's Hospital of Pittsburgh, and University of Pittsburgh School of Medicine, Pittsburgh, PA 15213.|Department of Surgery, University of Pittsburgh School of Medicine, Pittsburgh, PA, 15213.
Dai
Shipan
S
Division of Pediatric Surgery, Children's Hospital of Pittsburgh, and University of Pittsburgh School of Medicine, Pittsburgh, PA 15213.|Department of Surgery, University of Pittsburgh School of Medicine, Pittsburgh, PA, 15213.
Gribar
Steven C
SC
Division of Pediatric Surgery, Children's Hospital of Pittsburgh, and University of Pittsburgh School of Medicine, Pittsburgh, PA 15213.|Department of Surgery, University of Pittsburgh School of Medicine, Pittsburgh, PA, 15213.
Richardson
Ward
W
Division of Pediatric Surgery, Children's Hospital of Pittsburgh, and University of Pittsburgh School of Medicine, Pittsburgh, PA 15213.|Department of Surgery, University of Pittsburgh School of Medicine, Pittsburgh, PA, 15213.
Kohler
Jeff W
JW
Division of Pediatric Surgery, Children's Hospital of Pittsburgh, and University of Pittsburgh School of Medicine, Pittsburgh, PA 15213.|Department of Surgery, University of Pittsburgh School of Medicine, Pittsburgh, PA, 15213.
Hoffman
Rosemary A
RA
Department of Surgery, University of Pittsburgh School of Medicine, Pittsburgh, PA, 15213.
Branca
Maria F
MF
Division of Pediatric Surgery, Children's Hospital of Pittsburgh, and University of Pittsburgh School of Medicine, Pittsburgh, PA 15213.|Department of Surgery, University of Pittsburgh School of Medicine, Pittsburgh, PA, 15213.
Li
Jun
J
Division of Pediatric Surgery, Children's Hospital of Pittsburgh, and University of Pittsburgh School of Medicine, Pittsburgh, PA 15213.|Department of Surgery, University of Pittsburgh School of Medicine, Pittsburgh, PA, 15213.
Shi
Xiao-Hua
XH
Division of Pediatric Surgery, Children's Hospital of Pittsburgh, and University of Pittsburgh School of Medicine, Pittsburgh, PA 15213.|Department of Surgery, University of Pittsburgh School of Medicine, Pittsburgh, PA, 15213.
Sodhi
Chhinder P
CP
Division of Pediatric Surgery, Children's Hospital of Pittsburgh, and University of Pittsburgh School of Medicine, Pittsburgh, PA 15213.|Department of Surgery, University of Pittsburgh School of Medicine, Pittsburgh, PA, 15213.
Hackam
David J
DJ
Division of Pediatric Surgery, Children's Hospital of Pittsburgh, and University of Pittsburgh School of Medicine, Pittsburgh, PA 15213.|Department of Surgery, University of Pittsburgh School of Medicine, Pittsburgh, PA, 15213.
D000818:Animals; D002460:Cell Line; D018031:Connexin 43 / Q000096:biosynthesis / Q000172:deficiency / Q000235:genetics / Q000502:physiology*; D004927:Escherichia coli Infections / Q000276:immunology* / Q000401:mortality* / Q000473:pathology; D005260:Female; D006367:HeLa Cells; D006801:Humans; D008099:Liver / Q000166:cytology / Q000196:embryology / Q000276:immunology; D008264:Macrophages / Q000276:immunology* / Q000382:microbiology* / Q000473:pathology; D017737:Macrophages, Peritoneal / Q000276:immunology / Q000382:microbiology / Q000473:pathology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D010538:Peritonitis / Q000276:immunology* / Q000401:mortality* / Q000473:pathology; D010588:Phagosomes / Q000276:immunology / Q000378:metabolism / Q000382:microbiology; D016019:Survival Analysis
D018031:Connexin 43