Anti-beta2GPI-antibody-induced endothelial cell gene expression profiling reveals induction of novel pro-inflammatory genes potentially involved in primary antiphospholipid syndrome.
OBJECTIVE
To determine the effects of primary antiphospholipid syndrome (PAPS)-derived anti-beta(2)GPI antibodies on gene expression in human umbilical vein endothelial cells (HUVEC) by gene profiling using microarrays.


METHODS
Anti-beta(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-beta(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, IL1beta 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-beta(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-beta(2)GPI antibody-regulated genes could contribute to the vasculopathy associated with this disease.
Hamid
C
C
Research Institute of Healthcare Sciences, University of Wolverhampton, UK.
Norgate
K
K
D'Cruz
D P
DP
Khamashta
M A
MA
Arno
M
M
Pearson
J D
JD
Frampton
G
G
Murphy
J J
JJ
D000328:Adult; D000937:Antigen-Antibody Reactions; D016736:Antiphospholipid Syndrome / Q000276:immunology*; D001323:Autoantibodies / Q000276:immunology* / Q000494:pharmacology; D016022:Case-Control Studies; D015536:Down-Regulation; D019040:E-Selectin / Q000032:analysis; D042783:Endothelial Cells / Q000276:immunology*; D005260:Female; D020869:Gene Expression Profiling; D006801:Humans; D007074:Immunoglobulin G / Q000494:pharmacology; D016209:Interleukin-8 / Q000032:analysis; D008875:Middle Aged; D020411:Oligonucleotide Array Sequence Analysis; D053482:beta 2-Glycoprotein I / Q000276:immunology*
D001323:Autoantibodies; D019040:E-Selectin; D007074:Immunoglobulin G; D016209:Interleukin-8; D053482:beta 2-Glycoprotein I