Expression, function and cooperating partners of protease-activated receptor type 3 in vascular endothelial cells and B lymphocytes studied with specific monoclonal antibody

Olena Kalashnyk, Yuliya Petrova, Olena Lykhmus, Lyubov Mikhalovska, Sergiy Mikhalovsky, Anastasiya Zhukova, Dmitri Gnatenko, Wadie Bahou, Sergiy Komisarenko, Maryna Skok

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Receptor-specific antibodies can both prevent ligand-receptor interaction and initiate receptor signaling. Previously we generated monoclonal antibody 8E8 (mAb 8E8) against protease-activated receptor type 3 (PAR3) which inhibited proliferation of B cell hybridoma. Here we used mAb 8E8 and PAR1-specific polyclonal antibody to reveal the functions and cooperating partners of PAR3 in endothelial cells and in B lymphocytes. MAb 8E8 or PAR1 agonist peptide stimulated IL-6 and IL-8 production and VCAM-1 expression in HPMEC-ST1.6R cells. PAR1 antibody stimulated only VCAM-1 expression, while ICAM-1 expression was stimulated with mAB 8E8 or PAR3 peptide. MAb 8E8 stimulated weak mitogenic response, while PAR1 antibody inhibited it in normal but not in malignant B lymphocytes. Sandwich ELISA assay demonstrated the interaction of PAR3 with PAR1 in malignant cell lines and with IgM in normal B lymphocytes. It is concluded that PAR3 cooperates with PAR1 to mediate the effect of thrombin on cytokine production and VCAM-1 expression in endothelial cells and on cell proliferation in malignant B cells. ICAM-1 expression in endothelial cells requires PAR3 without PAR1. The inhibitory effect of thrombin in normal B lymphocytes is mediated by PAR1 alone, while mitogenic and pro-survival signaling in B lymphocytes is provided through PAR3 in cooperation with BCR.

Original languageEnglish
Pages (from-to)319-326
Number of pages8
JournalMolecular Immunology
Volume54
Issue number3-4
DOIs
Publication statusPublished - Jul 2013
Externally publishedYes

Fingerprint

B-Lymphocytes
Endothelial Cells
Monoclonal Antibodies
Vascular Cell Adhesion Molecule-1
Antibodies
Intercellular Adhesion Molecule-1
Thrombin
Peptides
Hybridomas
protease-activated receptor 3
Interleukin-8
Immunoglobulin M
Interleukin-6
Enzyme-Linked Immunosorbent Assay
Cell Proliferation
Cytokines
Ligands
Cell Line

Keywords

  • Antibodies
  • B lymphocytes
  • Endothelial cells
  • Protease-activated receptor
  • Thrombin

ASJC Scopus subject areas

  • Molecular Biology
  • Immunology

Cite this

Expression, function and cooperating partners of protease-activated receptor type 3 in vascular endothelial cells and B lymphocytes studied with specific monoclonal antibody. / Kalashnyk, Olena; Petrova, Yuliya; Lykhmus, Olena; Mikhalovska, Lyubov; Mikhalovsky, Sergiy; Zhukova, Anastasiya; Gnatenko, Dmitri; Bahou, Wadie; Komisarenko, Sergiy; Skok, Maryna.

In: Molecular Immunology, Vol. 54, No. 3-4, 07.2013, p. 319-326.

Research output: Contribution to journalArticle

Kalashnyk, O, Petrova, Y, Lykhmus, O, Mikhalovska, L, Mikhalovsky, S, Zhukova, A, Gnatenko, D, Bahou, W, Komisarenko, S & Skok, M 2013, 'Expression, function and cooperating partners of protease-activated receptor type 3 in vascular endothelial cells and B lymphocytes studied with specific monoclonal antibody', Molecular Immunology, vol. 54, no. 3-4, pp. 319-326. https://doi.org/10.1016/j.molimm.2012.12.021
Kalashnyk, Olena ; Petrova, Yuliya ; Lykhmus, Olena ; Mikhalovska, Lyubov ; Mikhalovsky, Sergiy ; Zhukova, Anastasiya ; Gnatenko, Dmitri ; Bahou, Wadie ; Komisarenko, Sergiy ; Skok, Maryna. / Expression, function and cooperating partners of protease-activated receptor type 3 in vascular endothelial cells and B lymphocytes studied with specific monoclonal antibody. In: Molecular Immunology. 2013 ; Vol. 54, No. 3-4. pp. 319-326.
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AU - Mikhalovsky, Sergiy

AU - Zhukova, Anastasiya

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