Detection of the "Kreisler" (maf B) gene by combination of in situ-hybridization and immunohistochemistry of RA-, osteoarthritis- and normal controls-synovial tissue samplings as a potential significant marker for early RA

Introduction

By analyzing gene expression profiles of arthritic tissue on DNA microarrays (EOS) compared to the "Body Atlas", a reference database of 13 normal human tissues, we found the RAB3 "Kreisler" (maf B) gene (member of the maf gene family and encoding for a transcription activator specific for mesenchymal and neuronal organogenesis) highly expressed in early rheumatoid arthritis (RA) (< 2 years disease duration).

Objective

To investigate the functional "Kreisler" gene expression in RA-, osteoarthritis- and normal controls- synovial samplings by combination of in situ-hybridization and immunohistochemistry.

Methods

We analyzed synovial biopsies (n = 12; 7f/5m) from 5 RA- (3 early RA, 2 RA), 4 osteoarthritis-patients and 3 normal controls, which were taken by arthrotomy by various indications and miniarthroscopy of MCPs (2 early RA). Samples were analyzed by in situ-hybridization with the "Kreisler" (maf B) gene-mRNA and immunohistochemistry (e.g. Ki 67, CD 68).

Results

We detected increased "Kreisler"-mRNA levels in 3 early RA samples in the synovial lining layer and no signals in the control and compared samples. At higher concentrations (>1 ng/μl) of RNA-oligonucleotides unspecific hybridization-signals prevailed in tissues of all diseases (even in normal controls). The combination of both methods (in situ-hybridization and immunohistochemistry) identifies the single cells inside the synovial lining layer which contains the highly expressed RAB3 "Kreisler" (maf B) gene.

Conclusion

Based on the gene expression profiles through oligonucleotid-microchip-array-analysis by EOS and the detection of the increased "Kreisler" (maf B) gene expression in combination of in situ-hybridization and immunohistochemistry of RA-synovial tissue samplings we discuss the "Kreisler"-gene as a potential inducing element in the pathogenesis of early RA. Further serial studies are needed to clarify the significance of "Kreisler" especially for early RA and the molecular pathogenesis of this disease.