Anti-beta2GPI-antibody-induced endothelial cell gene expression profiling reveals induction of novel pro-inflammatory genes potentially involved in primary antiphospholipid syndrome
| Autor: | Colleen Hamid, David D'Cruz, Geoffrey Frampton, Munther A. Khamashta, matthew Arno, Jeremy D. Pearson, Kirstie Norgate, John J. Murphy |
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| Rok vydání: | 2007 |
| Předmět: |
Adult
Microarray Immunology Down-Regulation General Biochemistry Genetics and Molecular Biology Antigen-Antibody Reactions Rheumatology Gene expression Immunology and Allergy Beta 2-Glycoprotein I Humans Gene Autoantibodies Oligonucleotide Array Sequence Analysis biology Microarray analysis techniques Gene Expression Profiling Tenascin C Interleukin-8 Endothelial Cells Middle Aged Antiphospholipid Syndrome Molecular biology Gene expression profiling Extended Report beta 2-Glycoprotein I Case-Control Studies Immunoglobulin G biology.protein Female DNA microarray E-Selectin |
| Zdroj: | Annals of the rheumatic diseases. 66(8) |
| ISSN: | 0003-4967 |
| Popis: | Objective: To determine the effects of primary antiphospholipid syndrome (PAPS)-derived anti-β 2 GPI antibodies on gene expression in human umbilical vein endothelial cells (HUVEC) by gene profiling using microarrays. Methods: Anti-β 2 GPI antibodies purified from sera of patients with PAPS or control IgG isolated from normal subjects were incubated with HUVEC for 4 h before isolation of RNA and processing for hybridisation to Affymetrix Human Genome U133A-2.0 arrays. Data were analysed using a combination of the MAS 5.0 (Affymetrix) and GeneSpring (Agilent) software programmes. For selected genes microarray data were confirmed by real-time PCR analysis or at the protein level by ELISA. Results: A total of 101 genes were found to be upregulated and 14 genes were downregulated twofold or more in response to anti-β 2 GPI antibodies. A number of novel genes not previously associated with APS were induced, including chemokines CCL20, CXCL3, CX3CL1, CXCL5, CXCL2 and CXCL1, the receptors Tenascin C, OLR1, IL-18 receptor 1, and growth factors CSF2, CSF3 IL-6, IL1β and FGF18. The majority of downregulated genes were transcription factors/signalling molecules including ID2. Quantitative real-time RT-PCR analysis confirmed the microarray results for selected genes (CSF3, CX3CL1, FGF18, ID2, SOD2, Tenascin C). Conclusions: This study reveals a complex gene expression response in HUVEC to anti-β 2 GPI antibodies with multiple chemokines, pro-inflammatory cytokines, pro-thrombotic and pro-adhesive genes regulated by these antibodies in vitro . Some of these newly identified anti-β 2 GPI antibody-regulated genes could contribute to the vasculopathy associated with this disease. |
| Databáze: | OpenAIRE |
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