Rapid access of antibodies to alpha5beta1 integrin overexpressed on the luminal surface of tumor blood vessels.
Integrin alpha(5)beta(1) is overexpressed on endothelial cells of tumor vessels and is uniformly and rapidly accessible to antibodies in the bloodstream. Here, we determined whether antibodies rapidly gain access to integrin overexpressed on the abluminal (basolateral) surface of endothelial cells through vascular leakiness or whether the rapid accessibility results instead because the integrin is overexpressed on the luminal (apical) surface of endothelial cells due to loss of cell polarity. Using tumors in RIP-Tag2 transgenic mice as a model, we first compared the binding pattern of intravascular anti-alpha(5)beta(1) integrin antibody with the leakage pattern of nonspecific IgG. The distributions did not match: anti-alpha(5)beta(1) integrin antibody uniformly labeled the tumor vasculature, but IgG was located in patchy sites of leakage. We next injected an antibody to fibrinogen/fibrin, which resulted in patchy labeling of tumors that matched the leakage of IgG and the overall distribution of fibrin in tumors. Similarly, injected antibodies to the basement membrane protein fibronectin, a ligand of alpha(5)beta(1) integrin, or type IV collagen produced patchy sites of leakage instead of uniform labeling of vascular basement membrane. Differences in the kinetics of labeling, which for alpha(5)beta(1) integrin antibody was near maximal by 10 minutes but for the other antibodies gradually increased over 6 hours, indicated differences in accessibility of their respective targets. Isosurface rendering of confocal microscopic images was consistent with antibody binding to alpha(5)beta(1) integrin on the luminal surface of endothelial cells. Together, these findings indicate that the rapid accessibility of alpha(5)beta(1) integrin in RIP-Tag2 tumors results from overexpression of the integrin on the luminal surface of tumor vessels.
Magnussen
Anette
A
Cardiovascular Research Institute, Comprehensive Cancer Center, and Department of Anatomy, University of California, San Francisco, California 94143-0130, USA.
Kasman
Ian M
IM
Norberg
Scott
S
Baluk
Peter
P
Murray
Richard
R
McDonald
Donald M
DM
D007516:Adenoma, Islet Cell / Q000098:blood supply*; D000818:Animals; D000906:Antibodies / Q000008:administration & dosage / Q000276:immunology / Q000378:metabolism*; D001808:Blood Vessels / Q000276:immunology / Q000378:metabolism; D024141:Collagen Type IV / Q000378:metabolism; D042783:Endothelial Cells / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D005337:Fibrin / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism; D007074:Immunoglobulin G / Q000008:administration & dosage / Q000276:immunology / Q000378:metabolism; D039081:Integrin alpha5beta1 / Q000096:biosynthesis / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009389:Neovascularization, Pathologic / Q000276:immunology / Q000378:metabolism; D010190:Pancreatic Neoplasms / Q000098:blood supply*; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000276:immunology; D014018:Tissue Distribution
D000906:Antibodies; D024141:Collagen Type IV; D005353:Fibronectins; D007074:Immunoglobulin G; D039081:Integrin alpha5beta1; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D005337:Fibrin