Binding of FimD on Bordetella pertussis to very late antigen-5 on monocytes activates complement receptor type 3 via protein tyrosine kinases.
Nonopsonized Bordetella pertussis bind to human monocytes by means of the virulence factors filamentous hemagglutinin (FHA), pertactin, and the minor fimbrial subunit FimD. Receptors on monocytes that mediate binding of B. pertussis to these cells include complement receptor type 3 (CR3), which binds to FHA of B. pertussis, and very late antigen-5 (VLA-5), which binds to an, as yet, unknown ligand on these bacteria. In the present study, the possibility that FimD acts as a ligand for VLA-5 was investigated. Soluble fibronectin, which is the natural ligand for VLA-5, or mAbs against VLA-5 inhibited binding to monocytes of B. pertussis strains that express FimD but not of mutant strains that lack FimD. Beads that were coated with the fusion protein maltose-binding protein-FimD bound to adherent monocytes, and this binding was inhibited by soluble fibronectin or mAb against the alpha- or beta-chain of VLA-5, while soluble collagen or mAb against VLA-4, VLA-6, CR3, or HLA class II had no effect. Down-modulation of VLA-5 on the apical surface of monocytes by plating the cells onto surfaces precoated with anti-VLA-5 mAb also inhibited binding of beads coated with maltose-binding protein-FimD to monocytes, while precoating of the surfaces with mAb against VLA-6 or CR3 had no effect. These results indicate that VLA-5 on monocytes serves as a receptor for FimD on B. pertussis. Binding of C3bi-coated erythrocytes to monocytes, which is a measure of the binding activity of CR3, was enhanced when monocytes were adhered onto plates precoated with purified fimbriae of B. pertussis, while precoating with fimbriae lacking FimD had no effect. Precoating of the plates with FimD-containing fimbriae also enhanced binding of B. pertussis, which express FHA, but not of strains that lack FHA, to monocytes. The enhanced binding of C3bi-coated erythrocytes and B. pertussis to monocytes could be markedly inhibited by tyrphostin-47, a protein tyrosine kinase inhibitor. These results demonstrate that interaction of FimD of B. pertussis with VLA-5 on monocytes activates CR3, which requires protein tyrosine kinases and results in enhanced binding of B. pertussis to the latter receptor via FHA.
Hazenbos
W L
WL
Department of Infectious Diseases, University Hospital, Leiden, The Netherlands.
van den Berg
B M
BM
Geuijen
C W
CW
Mooi
F R
FR
van Furth
R
R
D001422:Bacterial Adhesion / Q000276:immunology; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism; D001886:Bordetella pertussis / Q000737:chemistry / Q000276:immunology*; D002352:Carrier Proteins / Q000378:metabolism; D003167:Complement Activation / Q000276:immunology*; D036781:Fimbriae Proteins; D006801:Humans; D016177:Macrophage-1 Antigen / Q000378:metabolism*; D058619:Maltose-Binding Proteins; D009000:Monocytes / Q000201:enzymology / Q000378:metabolism*; D011485:Protein Binding / Q000276:immunology; D011505:Protein-Tyrosine Kinases / Q000276:immunology*; D017469:Receptors, Fibronectin / Q000378:metabolism*
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D002352:Carrier Proteins; D016177:Macrophage-1 Antigen; D058619:Maltose-Binding Proteins; D017469:Receptors, Fibronectin; C489284:fimD protein, Bordetella; C063629:fimD protein, bacteria; D036781:Fimbriae Proteins; D011505:Protein-Tyrosine Kinases