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Thromb Haemost 2016; 116(06): 1122-1130
DOI: 10.1160/TH16-04-0333
Blood Cells, Inflammation and Infection
Schattauer Publishers Schattauer

Anti-VLDL receptor monoclonal antibodies inhibit fibrin-VLDL receptor interaction and reduce fibrin-dependent leukocyte transmigration

Sergiy Yakovlev
1   Center for Vascular and Inflammatory Diseases, University of Maryland School of Medicine, Baltimore, Maryland, USA
2   Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, Maryland, USA
,
Alexey M. Belkin
1   Center for Vascular and Inflammatory Diseases, University of Maryland School of Medicine, Baltimore, Maryland, USA
2   Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, Maryland, USA
6   Present address: Center for Scientific Review, National Institutes of Health, Bethesda, Maryland, USA
,
Ling Chen
3   Department of Medicine, University of Maryland School of Medicine, Baltimore, Maryland, USA
4   Department of Physiology, University of Maryland School of Medicine, Baltimore, Maryland, USA
,
Chunzhang Cao
1   Center for Vascular and Inflammatory Diseases, University of Maryland School of Medicine, Baltimore, Maryland, USA
4   Department of Physiology, University of Maryland School of Medicine, Baltimore, Maryland, USA
,
Li Zhang
1   Center for Vascular and Inflammatory Diseases, University of Maryland School of Medicine, Baltimore, Maryland, USA
4   Department of Physiology, University of Maryland School of Medicine, Baltimore, Maryland, USA
,
Dudley K. Strickland
1   Center for Vascular and Inflammatory Diseases, University of Maryland School of Medicine, Baltimore, Maryland, USA
4   Department of Physiology, University of Maryland School of Medicine, Baltimore, Maryland, USA
5   Department of Surgery, University of Maryland School of Medicine, Baltimore, Maryland, USA
,
Leonid Medved
1   Center for Vascular and Inflammatory Diseases, University of Maryland School of Medicine, Baltimore, Maryland, USA
2   Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, Maryland, USA
› Author Affiliations Financial support: This work was supported by National Institutes of Health R01 Grants HL056051 to L. M., HL114379 and HL120388 to D. K.S, and NS082607 to L. Z.
Further Information

Publication History

Received: 26 April 2016

Accepted after major revision: 04 June 2016

Publication Date:
09 March 2018 (online)

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Summary

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.

Keywords

Fibrinogen/fibrin - VLDL receptor - leukocyte transmigration - monoclonal antibodies
 

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