SAT0031 Fine epitope mapping of immunodominant peptide epitopes by affinity matrix technology using yersinia-hsp60-specific synovial cd4+ t-cells in reactive arthritis (rea) patients. (1st June 2001)

• Record Type: Journal Article
• Authors: Thiel, A; Wu, P; Stauch, D; Rudwaleit, M; Radbruch, A; Sieper, J
• Is Part Of: Annals of the rheumatic diseases. Volume 60(2001)Supplement 1
• Page Start: A150
• Page End: A150
• Publication Date: 2001-06-01
• Subjects: Rheumatism -- Periodicals; 616.723005
• DOI: 10.1136/annrheumdis-2001.383
• Languages: English
• ISSNs: 0003-4967

Abstract

Abstract : Background: A T cell response to persitent bacterial antigen is present in patients with reactive arthritis (ReA). A better characterisation of this response might lead to new treatments. Objectives: To identify immundominant Yer-hsp60-derived CD4 T cell epitopes in Yersinia-induced reactive arthritis (ReA). Methods: In two patients with Yersinia-induced ReA synovial fluid (SF) T cell response were first investigated after in vitro stimulation with the Yersinia-derived protein hsp60. Fixed cells were stained with antibodies against CD4, CD8, CD69 and against the cytokine IFNgamma;. Patients with reacting CD4 + Th cells were retested and live IFNgamma-secreting cells were isolated with the affinity matrix technology after short term stimulation with Yer-hsp60. Cells were expanded within 2 to 5 weeks and retested with overlapping 18 mer peptides from the Yer-hsp60 protein performing cytometric analysis of fixed cells after short term stimulation with peptided pulsed autologous PBMCs. Results: Live antigen-specific IFNgamma;-secreting CD4 + T cells were enriched to a purity of 80 to 90%. After two weeks in vitro expansion still 80% of the short-term cultivated T cells reacted with IFNgamma-production upon Yer-hsp60 stimulation, while reaction upon stimulation with a irrelevant protein was below 1%. T cells were further expanded antigen-specifically for 3 weeks and their fine specificity was further mapped to one peptide in each patient (AA 139–156 and AA 7–30 of Yer-hsp60/GroEL). Tese peptides was also recognised by the patient's CD4 + T cells when whole blood or synovial fluid was stimulated for 6 h. Conclusion: This approach allows a sensitive and fast epitope mapping of immunodominant epitopes possibly involved in the pathogenesis of ReA.