Anti-β2-GPI Antibodies Induce Endothelial Cell Expression of Tissue Factor by LRP6 Signal Transduction Pathway Involving Lipid Rafts
In this study we analyzed whether anti-β2-GPI antibodies from patients with APS induce the endothelial cell expression of Tissue Factor (TF) by a LRP6 signal transduction pathway involving lipid rafts. HUVEC were stimulated with affinity purified anti-β2-GPI antibodies. Both LRP6 and β-catenin phosp...
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MDPI AG
2022-04-01
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| Online Access: | https://www.mdpi.com/2073-4409/11/8/1288 |
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| author | Gloria Riitano Antonella Capozzi Serena Recalchi Daniela Caissutti Agostina Longo Vincenzo Mattei Fabrizio Conti Roberta Misasi Tina Garofalo Maurizio Sorice Valeria Manganelli |
| author_facet | Gloria Riitano Antonella Capozzi Serena Recalchi Daniela Caissutti Agostina Longo Vincenzo Mattei Fabrizio Conti Roberta Misasi Tina Garofalo Maurizio Sorice Valeria Manganelli |
| author_sort | Gloria Riitano |
| collection | DOAJ |
| description | In this study we analyzed whether anti-β2-GPI antibodies from patients with APS induce the endothelial cell expression of Tissue Factor (TF) by a LRP6 signal transduction pathway involving lipid rafts. HUVEC were stimulated with affinity purified anti-β2-GPI antibodies. Both LRP6 and β-catenin phosphorylation, as well as TF expression, were evaluated by western blot. Results demonstrated that triggering with affinity purified anti-β2-GPI antibodies induced LRP6 phosphorylation with consequent β-catenin activation, leading to TF expression on the cell surface. Interestingly, the lipid rafts affecting agent methyl-β-cyclodextrin as well as the LRP6 inhibitor Dickkopf 1 (DKK1) partially reduced the anti-β2-GPI antibodies effect, indicating that the anti-β2-GPI effects on TF expression may depend on a signalling transduction pathway involving both lipid rafts and LRP6. An interaction between β2-GPI, LRP6 and PAR-2 within these microdomains was demonstrated by gradient fractionation and coimmunoprecipitation experiments. Thus, anti-β2-GPI antibodies react with their target antigen likely associated to LRP6 and PAR-2 within plasma membrane lipid rafts of the endothelial cell. Anti-β2-GPI binding triggers β-catenin phosphorylation, leading to a procoagulant phenotype characterized by TF expression. These findings deal with a novel signal transduction pathway which provides new insight in the APS pathogenesis, improving the knowledge of valuable therapeutic target(s). |
| first_indexed | 2024-03-09T11:01:54Z |
| format | Article |
| id | doaj.art-20ff817c154342ada706393bbed9fba0 |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-09T11:01:54Z |
| publishDate | 2022-04-01 |
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| record_format | Article |
| series | Cells |
| spelling | doaj.art-20ff817c154342ada706393bbed9fba02023-12-01T01:13:29ZengMDPI AGCells2073-44092022-04-01118128810.3390/cells11081288Anti-β2-GPI Antibodies Induce Endothelial Cell Expression of Tissue Factor by LRP6 Signal Transduction Pathway Involving Lipid RaftsGloria Riitano0Antonella Capozzi1Serena Recalchi2Daniela Caissutti3Agostina Longo4Vincenzo Mattei5Fabrizio Conti6Roberta Misasi7Tina Garofalo8Maurizio Sorice9Valeria Manganelli10Department of Experimental Medicine, “Sapienza” University of Rome, 00161 Rome, ItalyDepartment of Experimental Medicine, “Sapienza” University of Rome, 00161 Rome, ItalyDepartment of Experimental Medicine, “Sapienza” University of Rome, 00161 Rome, ItalyDepartment of Experimental Medicine, “Sapienza” University of Rome, 00161 Rome, ItalyDepartment of Experimental Medicine, “Sapienza” University of Rome, 00161 Rome, ItalyDepartment of Experimental Medicine, “Sapienza” University of Rome, 00161 Rome, ItalyRheumatology Unit, Department of Clinical Internal, Anesthesiological and Cardiovascular Sciences, “Sapienza” University of Rome, 00161 Rome, ItalyDepartment of Experimental Medicine, “Sapienza” University of Rome, 00161 Rome, ItalyDepartment of Experimental Medicine, “Sapienza” University of Rome, 00161 Rome, ItalyDepartment of Experimental Medicine, “Sapienza” University of Rome, 00161 Rome, ItalyDepartment of Experimental Medicine, “Sapienza” University of Rome, 00161 Rome, ItalyIn this study we analyzed whether anti-β2-GPI antibodies from patients with APS induce the endothelial cell expression of Tissue Factor (TF) by a LRP6 signal transduction pathway involving lipid rafts. HUVEC were stimulated with affinity purified anti-β2-GPI antibodies. Both LRP6 and β-catenin phosphorylation, as well as TF expression, were evaluated by western blot. Results demonstrated that triggering with affinity purified anti-β2-GPI antibodies induced LRP6 phosphorylation with consequent β-catenin activation, leading to TF expression on the cell surface. Interestingly, the lipid rafts affecting agent methyl-β-cyclodextrin as well as the LRP6 inhibitor Dickkopf 1 (DKK1) partially reduced the anti-β2-GPI antibodies effect, indicating that the anti-β2-GPI effects on TF expression may depend on a signalling transduction pathway involving both lipid rafts and LRP6. An interaction between β2-GPI, LRP6 and PAR-2 within these microdomains was demonstrated by gradient fractionation and coimmunoprecipitation experiments. Thus, anti-β2-GPI antibodies react with their target antigen likely associated to LRP6 and PAR-2 within plasma membrane lipid rafts of the endothelial cell. Anti-β2-GPI binding triggers β-catenin phosphorylation, leading to a procoagulant phenotype characterized by TF expression. These findings deal with a novel signal transduction pathway which provides new insight in the APS pathogenesis, improving the knowledge of valuable therapeutic target(s).https://www.mdpi.com/2073-4409/11/8/1288antiphospholipid syndromeβ2-GPI-glycoprotein ILRP-6Tissue Factorsignal transduction |
| spellingShingle | Gloria Riitano Antonella Capozzi Serena Recalchi Daniela Caissutti Agostina Longo Vincenzo Mattei Fabrizio Conti Roberta Misasi Tina Garofalo Maurizio Sorice Valeria Manganelli Anti-β2-GPI Antibodies Induce Endothelial Cell Expression of Tissue Factor by LRP6 Signal Transduction Pathway Involving Lipid Rafts Cells antiphospholipid syndrome β2-GPI-glycoprotein I LRP-6 Tissue Factor signal transduction |
| title | Anti-β2-GPI Antibodies Induce Endothelial Cell Expression of Tissue Factor by LRP6 Signal Transduction Pathway Involving Lipid Rafts |
| title_full | Anti-β2-GPI Antibodies Induce Endothelial Cell Expression of Tissue Factor by LRP6 Signal Transduction Pathway Involving Lipid Rafts |
| title_fullStr | Anti-β2-GPI Antibodies Induce Endothelial Cell Expression of Tissue Factor by LRP6 Signal Transduction Pathway Involving Lipid Rafts |
| title_full_unstemmed | Anti-β2-GPI Antibodies Induce Endothelial Cell Expression of Tissue Factor by LRP6 Signal Transduction Pathway Involving Lipid Rafts |
| title_short | Anti-β2-GPI Antibodies Induce Endothelial Cell Expression of Tissue Factor by LRP6 Signal Transduction Pathway Involving Lipid Rafts |
| title_sort | anti β2 gpi antibodies induce endothelial cell expression of tissue factor by lrp6 signal transduction pathway involving lipid rafts |
| topic | antiphospholipid syndrome β2-GPI-glycoprotein I LRP-6 Tissue Factor signal transduction |
| url | https://www.mdpi.com/2073-4409/11/8/1288 |
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