- Meeting abstract
- Open Access
T cell response in Shigella-induced reactive arthritis
Arthritis Res Ther volume 6, Article number: 39 (2004)
Shigella flexneri is one of the bacteria causing reactive arthritis (ReA). However, no target antigen for the cellular and humoral immune response against Shigella is yet known.
Shigella were grown in culture followed by differential centrifugation, after which a cytoplasmic and the 70S ribosomal fraction were obtained. By means of density gradient centrifugation the 70S ribosome was divided into its two subunits, the small ribosomal subunit 30S and the large ribosomal subunit 50S. The proteins derived from both subunits were further separated by FPLC–ion exchange chromatography. Mononuclear cells from synovial fluid of 3 HLA-B27 positive patients with Shigella-induced ReA were stimulated with whole Shigella, the cytoplasmic fraction, the ribosomal subunits and with single protein fractions from the ribosomal subunits gained by FPLC. Stimulatory protein fractions were subsequently cleaned by RP-HPLC and identified by stamp-size 2D PAGE.
Seventeen protein fractions from the 30S subunit and 23 protein fractions from the 50S subunit were obtained. A lymphocyte proliferation was seen after stimulation with whole Shigella, with the cytoplasmic fraction and the two ribosomal subunits but not with control antigens. More interestingly, five single ribosomal proteins out of 40 were equally stimulatory in all three patients, indicating that they represent immunodominant T cell antigens. Characterization by 2D PAGE identified the ribosomal proteins L15, L18, L21, L22 and L23 from 50S ('L' for large subunit) and S3 from the 30S ('S' for small subunit).
This approach allows the identification of T-cell epitopes derived from bacteria in ReA patients. In a first step we identified proteins from the highly conserved ribosomal fraction of Shigella flexneri. Their role in the pathogenesis must be further investigated.