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

3 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 1 2013

Keywords

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

ASJC Scopus subject areas

  • Immunology
  • Molecular Biology

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    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, 54(3-4), 319-326. https://doi.org/10.1016/j.molimm.2012.12.021