Objective: To identify genes that may participate in the pathophysiology of Sjögren's syndrome (SS), the technique of differential display was applied to labial minor salivary gland (MSG) biopsy samples.
Methods: Total RNA was isolated from MSG biopsy samples from a woman with primary SS and a control subject, and the differential display protocol with 8 different random oligonucleotide primers was performed. One particular differentially expressed fragment showed 98% homology with the cysteine-rich secretory protein 3 (CRISP-3) gene. The result was verified by reverse transcription-polymerase chain reaction (RT-PCR) with messenger RNA (mRNA) samples from MSG biopsy tissues obtained from 4 women with primary SS. A CRISP-3 RNA probe was synthesized for in situ hybridization of 7 MSG biopsy samples from patients with primary SS. In an attempt to interpret the expression of CRISP-3, normal peripheral blood lymphocytes (PBLs) were activated in vitro at different time points and assayed for CRISP-3 expression. Finally, B cells were transfected with the coding region of CRISP-3 and monitored for the up-regulation of different B cell activation markers.
Results: The CRISP-3 gene was detected by RT-PCR in all SS patients tested. Mainly the mononuclear cells infiltrating the MSGs of patients expressed CRISP-3 mRNA. In addition, CRISP-3 was detected by RT-PCR between 30 minutes and 6 hours in phorbol myristate acetate-activated normal PBLs, while staurosporine inhibited this expression. CRISP-3-transfected B cells exhibited an up-regulation in CD25 surface expression.
Conclusion: The CRISP-3 gene is identified as a novel early response gene that may participate in the pathophysiology of the autoimmune lesions of SS.