Peptide-mediated inhibition of neutrophil transmigration by blocking CD47 interactions with signal regulatory protein alpha.
CD47, a cell surface transmembrane Ig superfamily member, is an extracellular ligand for signal regulatory protein (SIRPalpha). Interactions between CD47 and SIRPalpha regulate many important immune cell functions including neutrophil (PMN) transmigration. Here we report identification of a novel function-blocking peptide, CERVIGTGWVRC, that structurally mimics an epitope on CD47 and binds to SIRPalpha. The CERVIGTGWVRC sequence was identified by panning phage display libraries on the inhibitory CD47 mAb, C5D5. In vitro PMN migration assays demonstrated that peptide CERVIGTGWVRC specifically inhibited PMN migration across intestinal epithelial monolayers and matrix in a dose-dependent fashion. Further studies using recombinant proteins indicated that the peptide specifically blocks CD47 and SIRPalpha binding in a dose-dependent fashion. Protein binding assays using SIRPalpha domain-specific recombinant proteins demonstrated that this peptide directly bound to the distal-most Ig loop of SIRPalpha, the same loop where CD47 binds. In summary, these findings support the relevance of CD47-SIRPalpha interactions in regulation of PMN transmigration and provide structural data predicting the key residues involved on the surface of CD47. Such peptide reagents may be useful for studies on experimental models of inflammation and provide a template for the design of anti-inflammatory agents.
Liu
Yuan
Y
Department of Pathology and Laboratory Medicine, Emory University, Atlanta, GA 30322, USA.
O'Connor
Miriam B
MB
Mandell
Kenneth J
KJ
Zen
Ke
K
Ullrich
Axel
A
Bühring
Hans-Jörg
HJ
Parkos
Charles A
CA
D000595:Amino Acid Sequence; D019138:Antibodies, Blocking / Q000378:metabolism; D000911:Antibodies, Monoclonal / Q000378:metabolism; D015703:Antigens, CD / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D000943:Antigens, Differentiation / Q000378:metabolism; D017104:Bacteriophage M13 / Q000276:immunology / Q000378:metabolism; D001666:Binding Sites, Antibody; D051928:CD47 Antigen; D002352:Carrier Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D002464:Cell Migration Inhibition; D006801:Humans; D007413:Intestinal Mucosa / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D008969:Molecular Sequence Data; D039842:Neural Cell Adhesion Molecule L1 / Q000037:antagonists & inhibitors* / Q000378:metabolism; D020556:Neutrophil Infiltration / Q000276:immunology*; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D019151:Peptide Library; D010449:Peptide Mapping / Q000379:methods; D011485:Protein Binding / Q000276:immunology; D017434:Protein Structure, Tertiary; D011971:Receptors, Immunologic / Q000037:antagonists & inhibitors* / Q000378:metabolism; D015398:Signal Transduction / Q000276:immunology*
D019138:Antibodies, Blocking; D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000943:Antigens, Differentiation; D051928:CD47 Antigen; C497894:CD47 protein, human; D002352:Carrier Proteins; D008562:Membrane Glycoproteins; D039842:Neural Cell Adhesion Molecule L1; D009842:Oligopeptides; D019151:Peptide Library; D011971:Receptors, Immunologic