Screening And Application Of New Autoantigens In Behcet’s Disease

Behcet’s disease (BD) is a multi-system inflammatory disease which might be involved in genetic factors, microbial infection and other living environmental triggers. There are some specific genetic loci which were found and reported in Asian populations with BD. However, the correlation with environmental microbial infection is still needed further research.Five different human original cell lines were investigated to detect serum autoantibody profiling in BD. Based on the significant differences of fluorescence intensities from five cell lines, human umbilical vein endothelial cell (HUVEC) was supposed to be the suitable target cell which contained potential antigens against autoantibodies in sera from BD patients. Then systems biotechnology, including western blot, immunoprecipitation,2-DE and individual proteomic microarray, were combined and used to screen the potential autoantigens in HUVEC. With the reaction to BD sera, two proteins were screened out and identified as prohibitin and β-tubulin by MALDI-TOF/TOF mass spectrometry in succession. After molecular construction, expression and purification, the recombinant prohibitin and P-tubulin were respectively used to comprise ELISA kits to test the prevalence of autoantibodies in patients’sera. As a result, the prevalence of serum antibodies against recombinant prohibitin or β-tubulin were all significant than healthy donors.In conclusion, the study reported two new autoantigens (prohibitin and β-tubulin) as potential antigens. And the correlation between these two autoantigens and microbial heat shock protein65suggested that microbes and patients’ habitats might be involved in the pathogenesis of BD.