The soluble variant surface glycoprotein of African trypanosomes is recognized by a macrophage scavenger receptor and induces I kappa B alpha degradation independently of TRAF6-mediated TLR signaling.
The GPI residues of soluble variant surface glycoprotein (sVSG) molecules released from the membrane of African trypanosomes during infection induce macrophage activation events. In this study, we demonstrate that the trypanosome sVSG molecule binds to the membrane of murine RAW 264.7 macrophages and activates the NF-kappaB cascade independently of a TLR-mediated interaction. The binding of fluorochrome-labeled sVSG molecules to macrophage membranes was saturable, was inhibited by the scavenger receptor-specific ligand maleylated BSA, and was followed by rapid intracellular uptake of the molecules and subsequent internalization to lysosomal compartments. Inhibition of cellular phagocytic and endocytic uptake processes by cytochalasin B and monodansylcadaverine, respectively, revealed that sVSG internalization was necessary for IkappaBalpha degradation and occurred by an actin-dependent, clathrin-independent process. Activation of RAW 264.7 cells by sVSG following treatment of the cells with the TRAF6 inhibitory peptide DIVK resulted in enhanced NF-kappaB signaling, suggesting both that TRAF6-dependent TLR activation of the pathway alone is not required for signaling and that TLR pathway components may negatively regulate expression of sVSG-induced signaling. These results demonstrate that stimulation of macrophages by sVSG involves a complex process of receptor-mediated binding and uptake steps, leading to both positive and negative signaling events that ultimately regulate cellular activation.
Leppert
Brian J
BJ
Department of Medical Microbiology and Immunology, School of Medicine and Public Health, University of Wisconsin, Madison, WI 53706.
Mansfield
John M
JM
Paulnock
Donna M
DM
D000199:Actins / Q000502:physiology; D000818:Animals; D002460:Cell Line; D002966:Clathrin / Q000502:physiology; D005260:Female; D020672:I-kappa B Proteins / Q000378:metabolism*; D008262:Macrophage Activation; D017737:Macrophages, Peritoneal / Q000276:immunology / Q000378:metabolism* / Q000469:parasitology; D051379:Mice; D008807:Mice, Inbred BALB C; D000072000:NF-KappaB Inhibitor alpha; D010587:Phagocytosis / Q000276:immunology; D051116:Receptors, Scavenger / Q000378:metabolism*; D015398:Signal Transduction / Q000276:immunology*; D012995:Solubility; D048029:TNF Receptor-Associated Factor 6 / Q000502:physiology*; D051193:Toll-Like Receptors / Q000502:physiology*; D016833:Trypanosoma brucei rhodesiense; D014643:Variant Surface Glycoproteins, Trypanosoma / Q000378:metabolism* / Q000502:physiology
D000199:Actins; D002966:Clathrin; D020672:I-kappa B Proteins; C000606205:Nfkbia protein, mouse; D051116:Receptors, Scavenger; D048029:TNF Receptor-Associated Factor 6; D051193:Toll-Like Receptors; D014643:Variant Surface Glycoproteins, Trypanosoma; D000072000:NF-KappaB Inhibitor alpha