Expression, function and cooperating partners of protease-activated receptor type 3 in vascular endothelial cells and B lymphocytes studied with specific monoclonal antibody
| Autor: | Olena Lykhmus, L. Mikhalovska, Wadie F. Bahou, Anastasiya I. Zhukova, Maryna Skok, Sergiy Mikhalovsky, Dmitri V. Gnatenko, Olena Kalashnyk, Yuliya I. Petrova, Serhiy Komisarenko |
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| Rok vydání: | 2012 |
| Předmět: |
Male
medicine.drug_class medicine.medical_treatment Immunology B-cell receptor Vascular Cell Adhesion Molecule-1 Monoclonal antibody Cell Line Mice Antibody Specificity medicine Animals Humans Protease-activated receptor Receptor PAR-1 Molecular Biology B cell Cell Proliferation B-Lymphocytes Mice Inbred BALB C biology Interleukin-6 Interleukin-8 Thrombin Antibodies Monoclonal Endothelial Cells Molecular biology B-1 cell Mice Inbred C57BL Cytokine medicine.anatomical_structure Polyclonal antibodies cardiovascular system biology.protein Receptors Thrombin Antibody |
| Zdroj: | Molecular immunology. 54(3-4) |
| ISSN: | 1872-9142 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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