Antibodies against the first Ig-like domain of human platelet endothelial cell adhesion molecule-1 (PECAM-1) that inhibit PECAM-1-dependent homophilic adhesion block in vivo neutrophil recruitment.
Platelet endothelial cell adhesion molecule (PECAM-1), a member of the Ig superfamily, is found on endothelial cells and neutrophils and has been shown to be involved in the migration of leukocytes across the endothelium. Adhesion is mediated, at least in part, through binding interactions involving its first N-terminal Ig-like domain, but it is still unclear which sequences in this domain are required for in vivo function. Therefore, to identify functionally important regions of the first Ig-like domain of PECAM-1 that are required for the participation of PECAM-1 in in vivo neutrophil recruitment, a panel of mAbs against this region of PECAM-1 was generated and characterized in in vitro adhesion assays and in an in vivo model of cutaneous inflammation. It was observed that mAbs that disrupted PECAM-1-dependent homophilic adhesion in an L cell aggregation assay also blocked TNF-alpha-induced intradermal accumulation of neutrophils in a transmigration model using human skin transplanted onto SCID mice. Localization of the epitopes of these Abs indicated that these function-blocking Abs mapped to specific regions on either face of domain 1. This suggests that these regions of the first Ig-like domain may contain or be close to binding sites involved in PECAM-1-dependent homophilic adhesion, and thus may represent potential targets for the development of antiinflammatory reagents.
Nakada
M T
MT
Centocor, Malvern, PA 19355, USA.
Amin
K
K
Christofidou-Solomidou
M
M
O'Brien
C D
CD
Sun
J
J
Gurubhagavatula
I
I
Heavner
G A
GA
Taylor
A H
AH
Paddock
C
C
Sun
Q H
QH
Zehnder
J L
JL
Newman
P J
PJ
Albelda
S M
SM
DeLisser
H M
HM
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000378:metabolism / Q000494:pharmacology*; D001666:Binding Sites, Antibody; D001667:Binding, Competitive; D002417:Cattle; D002448:Cell Adhesion / Q000276:immunology; D002449:Cell Aggregation / Q000276:immunology; D002465:Cell Movement / Q000276:immunology; D004730:Endothelium, Vascular / Q000276:immunology / Q000378:metabolism; D018604:Epitope Mapping; D006801:Humans; D007136:Immunoglobulins / Q000276:immunology*; D007739:L Cells; D051379:Mice; D016513:Mice, SCID; D008969:Molecular Sequence Data; D018375:Neutrophil Activation / Q000276:immunology*; D009504:Neutrophils / Q000276:immunology / Q000473:pathology; D010446:Peptide Fragments / Q000276:immunology*; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000276:immunology* / Q000378:metabolism / Q000502:physiology; D017434:Protein Structure, Tertiary; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D016038:Skin Transplantation / Q000473:pathology
D000911:Antibodies, Monoclonal; D007136:Immunoglobulins; D010446:Peptide Fragments; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D011993:Recombinant Fusion Proteins