Blood cell gene expression profiling in rheumatoid arthritis. Discriminative genes and effect of rheumatoid factor.
To study the pathogenic importance of the rheumatoid factor (RF) in rheumatoid arthritis (RA) and to identify genes differentially expressed in patients and healthy individuals, total RNA was isolated from peripheral blood mononuclear cells (PBMC) from eight RF-positive and six RF-negative RA patients, and seven healthy controls. Gene expression of about 10,000 genes were examined using oligonucleotide-based DNA chip microarrays. The analyses showed no significant differences in PBMC expression patterns from RF-positive and RF-negative patients. However, comparisons of gene expression patterns from all fourteen RA patients and healthy controls identified a subset of discriminative genes. These results were validated by real time reverse transcription polymerase chain reaction (RT-PCR) on another group of RA patients and healthy controls. This confirmed that the following genes had a significantly higher expression in RA patients than in healthy controls: CD14 antigen, defensin alpha-1 and alpha-3 (DEFA), fatty-acid-Coenzyme A ligase, long-chain 2 (FACL), ribonuclease 2 (RNASE2), S100 calcium-binding protein A8 and A12 (S100A8 and S100A12). In contrast, the expression of MHC class II, DQ beta1 (HLA-DQB1) was significantly reduced in RA patients compared to healthy controls.


CONCLUSIONS
With the analytical procedure employed, we did not find any indication that RF-positive and RF-negative RA are two fundamentally different diseases. Most of the genes discriminative between RA patients and healthy individuals are known to be involved in immunoinflammatory responses, especially those related to altered phagocytic functions.
Bovin
Lone Frier
LF
Institute for Inflammation Research IIR 7521, Rigshospitalet National University Hospital, Blegdamsvej 9, DK-2100, Copenhagen, Denmark.
Rieneck
Klaus
K
Workman
Christopher
C
Nielsen
Henrik
H
Sørensen
Søren Freiesleben
SF
Skjødt
Henrik
H
Florescu
Adrian
A
Brunak
Søren
S
Bendtzen
Klaus
K
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001172:Arthritis, Rheumatoid / Q000097:blood / Q000188:drug therapy / Q000235:genetics*; D001772:Blood Cell Count; D002097:C-Reactive Protein / Q000032:analysis; D003066:Coenzyme A Ligases / Q000235:genetics; D018076:DNA, Complementary / Q000138:chemical synthesis / Q000235:genetics; D003627:Data Interpretation, Statistical; D047208:Eosinophil-Derived Neurotoxin / Q000235:genetics; D005260:Female; D020869:Gene Expression Profiling; D006683:HLA-DQ Antigens / Q000235:genetics; D059866:HLA-DQ beta-Chains; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000378:metabolism*; D018950:Lipopolysaccharide Receptors / Q000235:genetics; D008297:Male; D008875:Middle Aged; D020411:Oligonucleotide Array Sequence Analysis; D025341:Principal Component Analysis; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012217:Rheumatoid Factor / Q000097:blood*; D009418:S100 Proteins / Q000235:genetics; D015854:Up-Regulation / Q000235:genetics; D023084:alpha-Defensins / Q000235:genetics; D000098859:Long-Chain-Fatty-Acid-CoA Ligase
D018076:DNA, Complementary; D006683:HLA-DQ Antigens; D059866:HLA-DQ beta-Chains; C069051:HLA-DQB1 antigen; D018950:Lipopolysaccharide Receptors; D009418:S100 Proteins; D023084:alpha-Defensins; D012313:RNA; D002097:C-Reactive Protein; D012217:Rheumatoid Factor; D047208:Eosinophil-Derived Neurotoxin; C479221:RNASE2 protein, human; D003066:Coenzyme A Ligases; D000098859:Long-Chain-Fatty-Acid-CoA Ligase