title: Anti-VLDL receptor monoclonal antibodies inhibit fibrin-VLDL receptor interaction and reduce fibrin-dependent leukocyte transmigration.
abstract: Our previous studies revealed that the interaction of fibrin with the very low density lipoprotein receptor (VLDLR) promotes transendothelial migration of leukocytes and thereby inflammation, and localised the fibrin-binding site to CR-domains 2-4 of this receptor. In the present study, we tested interaction of three anti-VLDLR monoclonal antibodies, mAb 1H10, 1H5, and 5F3, with recombinant fragments of VLDLR containing various combinations of its CR-domains and found that the epitopes for mAb 1H10 and mAb 1H5 overlap with the fibrin-binding site of VLDLR. Based on these findings, we hypothesised that mAb 1H10 and mAb 1H5 should inhibit fibrin-VLDLR interaction and modulate leukocyte transmigration. To test this hypothesis, we first demonstrated that these monoclonal antibodies both have high affinity to the fibrin-binding fragments of the VLDL receptor and efficiently inhibit interaction between the VLDLR-binding fragment of fibrin and the fibrin-binding fragments of VLDLR. Next, in the in vitro experiments using leukocyte transendothelial migration assay we found that both monoclonal antibodies efficiently inhibit leukocyte transmigration induced by fibrin mimetic NDSK-II. Finally, in vivo experiments using mouse model of peritonitis revealed that mAb 1H10 and mAb 1H5 both significantly reduce infiltration of leukocytes into the peritoneum. Furthermore, our experiments using mouse model of myocardial ischemia-reperfusion injury revealed that both monoclonal antibodies significantly reduce myocardial injury induced by ischaemia-reperfusion. Thus, the results obtained indicate that monoclonal antibodies 1H10 and 1H5 are novel specific inhibitors of fibrin-VLDLR-dependent leukocyte transmigration pathway. They may represent potential therapeutics for treatment of fibrin-dependent inflammation including myocardial ischaemia-reperfusion injury.
authors: Sergiy
authors: S
authors: Yakovlev
authors: Alexey M
authors: AM
authors: Belkin
authors: Ling
authors: L
authors: Chen
authors: Chunzhang
authors: C
authors: Cao
authors: Li
authors: L
authors: Zhang
authors: Dudley K
authors: DK
authors: Strickland
authors: Dr. Leonid Medved, PhD, University of Maryland School of Medicine, Center for Vascular and Inflammatory Diseases, 800 West Baltimore Street, Baltimore, MD, 21201, USA, Tel.: +1 410 706 8065, Fax: +1 410 706 8121, E-mail: Lmedved@som.umaryland.edu.
authors: Leonid
authors: L
authors: Medved
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D018604:Epitope Mapping; D005337:Fibrin / Q000378:metabolism*; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D007962:Leukocytes / Q000166:cytology*; D008810:Mice, Inbred C57BL; D011973:Receptors, LDL / Q000276:immunology* / Q000378:metabolism; D057705:Transendothelial and Transepithelial Migration
chemical_list: D000911:Antibodies, Monoclonal; D011973:Receptors, LDL; C036053:VLDL receptor; D005337:Fibrin
keywords: Fibrinogen/fibrin; VLDL receptor; leukocyte transmigration; monoclonal antibodies