TLR2 transmodulates monocyte adhesion and transmigration via Rac1- and PI3K-mediated inside-out signaling in response to Porphyromonas gingivalis fimbriae.
We present evidence for a novel TLR2 function in transmodulating the adhesive activities of human monocytes in response to the fimbriae of Porphyromonas gingivalis, a pathogen implicated in chronic periodontitis and atherosclerosis. Monocyte recruitment into the subendothelium is a crucial step in atherosclerosis, and we investigated the role of P. gingivalis fimbriae in stimulating monocyte adhesion to endothelial cells and transendothelial migration. Fimbriae induced CD11b/CD18-dependent adhesion of human monocytes or mouse macrophages to endothelial receptor ICAM-1; these activities were inhibited by TLR2 blockade or deficiency or by pharmacological inhibitors of PI3K. Moreover, this inducible adhesive activity was sensitive to the action of Clostridium difficile toxin B, but was not affected by Clostridium botulinum C3 exoenzyme, pertussis toxin, or cholera toxin. Accordingly, we subsequently showed through the use of dominant negative signaling mutants of small GTPases, that Rac1 mediates the ability of fimbria-stimulated monocytes to bind ICAM-1. A dominant negative mutant of Rac1 also inhibited the lipid kinase activity of PI3K suggesting that Rac1 acts upstream of PI3K in this proadhesive pathway. Furthermore, fimbriae stimulated monocyte adhesion to HUVEC and transmigration across HUVEC monolayers; both activities required TLR2 and Rac1 signaling and were dependent upon ICAM-1 and the high-affinity state of CD11b/CD18. P. gingivalis-stimulated monocytes displayed enhanced transendothelial migration compared with monocytes stimulated with nonfimbriated isogenic mutants. Thus, P. gingivalis fimbriae activate a novel proadhesive pathway in human monocytes, involving TLR2, Rac1, PI3K, and CD11b/CD18, which may constitute a mechanistic basis linking P. gingivalis to inflammatory atherosclerotic processes.
Harokopakis
Evlambia
E
Center for Oral Health and Systemic Disease, University of Louisville Health Sciences Center, KY 40292, USA.
Albzreh
Mohamad H
MH
Martin
Michael H
MH
Hajishengallis
George
G
D000818:Animals; D039481:CD11b Antigen / Q000378:metabolism / Q000502:physiology; D018821:CD18 Antigens / Q000378:metabolism / Q000502:physiology; D002448:Cell Adhesion / Q000276:immunology; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000276:immunology*; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000276:immunology / Q000382:microbiology; D005340:Fibrinogen / Q000378:metabolism; D010861:Fimbriae, Bacterial / Q000276:immunology / Q000502:physiology*; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D008024:Ligands; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009000:Monocytes / Q000166:cytology / Q000276:immunology* / Q000378:metabolism / Q000382:microbiology; D037342:Pertussis Toxin / Q000276:immunology; D019869:Phosphatidylinositol 3-Kinases / Q000502:physiology*; D016966:Porphyromonas gingivalis / Q000276:immunology / Q000502:physiology*; D015398:Signal Transduction / Q000276:immunology*; D051195:Toll-Like Receptor 2 / Q000172:deficiency / Q000235:genetics / Q000502:physiology*; D020830:rac1 GTP-Binding Protein / Q000502:physiology*
D039481:CD11b Antigen; D018821:CD18 Antigens; D008024:Ligands; D051195:Toll-Like Receptor 2; D018799:Intercellular Adhesion Molecule-1; D005340:Fibrinogen; D037342:Pertussis Toxin; D019869:Phosphatidylinositol 3-Kinases; D020830:rac1 GTP-Binding Protein