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Anti-β2GPI-antibody-induced endothelial cell gene expression profiling reveals induction of novel pro- inflammatory genes potentially involved in primary anti- phospholipid syndrome.
  1. C Hamid (colleen.hamid{at}kcl.ac.uk)
  1. Research Institute of Healthcare Sciences, University of Wolverhampton, United Kingdom
    1. K Norgate (kirstie.norgate{at}kcl.ac.uk)
    1. Division of Immunology Infection and Inflammatory Disease, King's College London, United Kingdom
      1. D P D'Cruz (david.d'cruz{at}kcl.ac.uk)
      1. Lupus Research Unit, St Thomas' Hospital, United Kingdom
        1. M A Khamashta (munther.khamashta{at}kcl.ac.uk)
        1. Lupus Research Unit, St Thomas' Hospital, United Kingdom
          1. M Arno (matthew.arno{at}kcl.ac.uk)
          1. Dept. of Nutrition and Dietetics, King's College London, United Kingdom
            1. J D Pearson (jeremy.pearson{at}kcl.ac.uk)
            1. Cardiovascular Division, King's College London, United Kingdom
              1. G Frampton (g.frampton{at}wlv.ac.uk)
              1. Research Institute of Healthcare Sciences, University of Wolverhampton, United Kingdom
                1. J J Murphy (john.murphy{at}kcl.ac.uk)
                1. Division of Immunology Infection and Inflammatory Disease, King's College London, United Kingdom

                  Abstract

                  Objectives: To determine the effects of primary anti-phospholipid syndrome (PAPS)-derived anti-β 2GPI antibodies on gene expression in human umbilical vein endothelial cells (HUVEC) by gene profiling using microarrays.

                  Methods: Anti-β2GPI antibodies purified from sera of patients with PAPS or control IgG isolated from normal subjects were incubated with HUVEC for four hours before isolation of RNA and processing for hybridization to Affymetrix Human Genome U133A-2.0 arrays. Data were analyzed 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 two fold or more in response to anti-β2GPI 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/signaling 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-β2GPI 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-β2GPI antibody-regulated genes could contribute to the vasculopathy associated with this disease.

                  • anti-beta2GPI antibodies
                  • anti-phospholipid syndrome
                  • microarray

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