{"id":{"repo_id":"wlv","oai_identifier":"oai:wlv.openrepository.com:2436/14407"},"canonical_url":"https://search.dev.ndltd.org/etd/wlv/oai:wlv.openrepository.com:2436/14407","repository":{"repo_id":"wlv","name":"University of Wolverhampton","base_url":"https://wlv.openrepository.com/server/oai/request"},"display":{"title":"Identification of Anti-Beta2 Glycoprotein I Auto-antibody Regulated Gene Targets in the Primary Antiphospholipid Syndrome Using Gene Microarray Analysis","abstract":"Anti-Beta2-Glycoprotein I antibodies (anti-2GPI) are strongly associated with thrombosis in patients with primary antiphospholipid syndrome (PAPS). Anti-2GPI activate endothelial cells (EC) resulting in a pro-thrombotic and pro-inflammatory phenotype. In order to characterise EC gene regulation in response to anti-2GPI, early global gene expression was assessed in human umbilical vein endothelial cells (HUVEC) in response to affinity purified anti-2GPI. Sera were collected from patients with PAPS and IgG was purified using HiTrap Protein G Sepharose columns. Polyclonal anti-2GPI were prepared by passing patient IgG through NHS activated sepharose coupled to human 2GPI. Anti-2GPI preparations were characterized by confirming their 2GPI co-factor dependence, binding to 2GPI and ability to induce leukocyte adhesion molecule expression and IL-8 production in vitro. Two microarray experiments tested differential global gene expression in 6 individual HUVEC donors in response to 5 different PAPS polyclonal anti-2GPI (50 mg/ml) compared to 5 normal control IgG (50 mg/ml) after 4 hours incubation . Total HUVEC RNA was extracted and cRNA was prepared and hybridised to Affymetrix HG-133A (Exp.1) and HG-133A_2 (Exp.2) gene chips. Data were analyzed using a combination of the MAS 5.0 (Affymetrix) and GeneSpring (Agilent) software programmes. Significant change in gene expression was defined as greater than two fold increase or decrease in expression (p<0.05). Novel genes not previously associated with PAPS 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, IL1 and FGF18. Downregulated genes were transcription factors/signaling molecules including ID2. Microarray results were confirmed for selected genes (CSF3, CX3CL1, FGF18, ID2, SOD2, Tenascin C) using quantitative real-time RT-PCR analysis. This study revealed a complex anti-2GPI-regulated gene expression profile in HUVEC in vitro. The novel chemokines and pro-inflammatory cytokines identified in this study may contribute to the vasculopathy associated with PAPS.","abstract_html":"Anti-Beta2-Glycoprotein I antibodies (anti-2GPI) are strongly associated with thrombosis in patients with primary antiphospholipid syndrome (PAPS). Anti-2GPI activate endothelial cells (EC) resulting in a pro-thrombotic and pro-inflammatory phenotype. In order to characterise EC gene regulation in response to anti-2GPI, early global gene expression was assessed in human umbilical vein endothelial cells (HUVEC) in response to affinity purified anti-2GPI. Sera were collected from patients with PAPS and IgG was purified using HiTrap Protein G Sepharose columns. Polyclonal anti-2GPI were prepared by passing patient IgG through NHS activated sepharose coupled to human 2GPI. Anti-2GPI preparations were characterized by confirming their 2GPI co-factor dependence, binding to 2GPI and ability to induce leukocyte adhesion molecule expression and IL-8 production in vitro. Two microarray experiments tested differential global gene expression in 6 individual HUVEC donors in response to 5 different PAPS polyclonal anti-2GPI (50 mg/ml) compared to 5 normal control IgG (50 mg/ml) after 4 hours incubation . Total HUVEC RNA was extracted and cRNA was prepared and hybridised to Affymetrix HG-133A (Exp.1) and HG-133A_2 (Exp.2) gene chips. Data were analyzed using a combination of the MAS 5.0 (Affymetrix) and GeneSpring (Agilent) software programmes. Significant change in gene expression was defined as greater than two fold increase or decrease in expression (p&lt;0.05). Novel genes not previously associated with PAPS 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, IL1 and FGF18. Downregulated genes were transcription factors/signaling molecules including ID2. Microarray results were confirmed for selected genes (CSF3, CX3CL1, FGF18, ID2, SOD2, Tenascin C) using quantitative real-time RT-PCR analysis. This study revealed a complex anti-2GPI-regulated gene expression profile in HUVEC in vitro. The novel chemokines and pro-inflammatory cytokines identified in this study may contribute to the vasculopathy associated with PAPS.","abstract_has_math":false,"creators":["Hamid, Colleen G."],"institution":"University of Wolverhampton","degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":[],"advisors":["Frampton, G.","Murphy, John"],"committee_chairs":[],"committee_members":[],"year":2007,"date_issued":"2007-10","date_published":"2007-10","updated_at":"2026-07-24T06:10:04Z","subjects":["Anti-β2 Glycoprotein I antibodies","Endothelial cells","Thrombosis","Primary antiphospholipid syndrome","Antiphospholipid antibodies"],"languages":[],"rights":[],"rights_urls":["https://wlv.dspace7.openrepository.com/bitstreams/68301682-f752-40f0-a5c0-3046bf536336/download"],"identifier_entries":[]},"links":{"outbound_url":null,"outbound_label":null,"outbound_source":null},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.advisor","label":"Advisor","values":["Frampton, G.","Murphy, John"]},{"key":"dc:creator","label":"Author","values":["Hamid, Colleen G."]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.issued","label":"Date","values":["2007-10"]},{"key":"dc:publisher.institution","label":"Dc Publisher Institution","values":["University of Wolverhampton"]},{"key":"dc:relation.isreferencedby","label":"Dc Relation Isreferencedby","values":["http://hdl.handle.net/2436/14407"]},{"key":"dc:type","label":"Dc Type","values":["Thesis or dissertation"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Anti-β2 Glycoprotein I antibodies","Endothelial cells","Thrombosis","Primary antiphospholipid syndrome","Antiphospholipid antibodies"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:rights","label":"Dc Rights","values":["https://wlv.dspace7.openrepository.com/bitstreams/68301682-f752-40f0-a5c0-3046bf536336/download"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.uri","label":"Identifier URI","values":["https://wlv.dspace7.openrepository.com/bitstreams/35af9829-d6c6-4d34-953c-d86e30fc4bc0/download"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["Anti-Beta2-Glycoprotein I antibodies (anti-2GPI) are strongly associated with thrombosis in patients with primary antiphospholipid syndrome (PAPS). 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