title: Use of HLA-B27 tetramers to identify low-frequency antigen-specific T cells in Chlamydia-triggered reactive arthritis.
abstract: Reports of the use of HLA-B27/peptide tetrameric complexes to study peptide-specific CD8+ T cells in HLA-B27+-related diseases are rare. To establish HLA-B27 tetramers we first compared the function of HLA-B27 tetramers with HLA-A2 tetramers by using viral epitopes. HLA-B27 and HLA-A2 tetramers loaded with immunodominant peptides from Epstein-Barr virus were generated with comparable yields and both molecules detected antigen-specific CD8+ T cells. The application of HLA-B27 tetramers in HLA-B27-related diseases was performed with nine recently described Chlamydia-derived peptides in synovial fluid and peripheral blood, to examine the CD8+ T cell response against Chlamydia trachomatis antigens in nine patients with Chlamydia-triggered reactive arthritis (Ct-ReA). Four of six HLA-B27+ Ct-ReA patients had specific synovial T cell binding to at least one HLA-B27/Chlamydia peptide tetramer. The HLA-B27/Chlamydia peptide 195 tetramer bound to synovial T cells from three of six patients and HLA-B27/Chlamydia peptide 133 tetramer to synovial T cells from two patients. However, the frequency of these cells was low (0.02-0.09%). Moreover, we demonstrate two methods to generate HLA-B27-restricted T cell lines. First, HLA-B27 tetramers and magnetic beads were used to sort antigen-specific CD8+ T cells. Second, Chlamydia-infected dendritic cells were used to stimulate CD8+ T cells ex vivo. Highly pure CD8 T cell lines could be generated ex vivo by magnetic sorting by using HLA-B27 tetramers loaded with an EBV peptide. The frequency of Chlamydia-specific, HLA-B27 tetramer-binding CD8+ T cells could be increased by stimulating CD8+ T cells ex vivo with Chlamydia-infected dendritic cells. We conclude that HLA-B27 tetramers are a useful tool for the detection and expansion of HLA-B27-restricted CD8+ T cells. T cells specific for one or more of three Chlamydia-derived peptides were found at low frequency in synovial fluid from HLA-B27+ patients with Ct-ReA. These cells can be expanded ex vivo, suggesting that they are immunologically functional.
authors.affiliation: Charite Berlin, Campus Benjamin Franklin, Department for Gastroenterology, Infectiology and Rheumatology, Berlin, Germany. heinerappel@yahoo.com
authors.forename: Heiner
authors.initials: H
authors.lastname: Appel
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Kuon
authors.forename: Maren
authors.initials: M
authors.lastname: Kuhne
authors.forename: Peihua
authors.initials: P
authors.lastname: Wu
authors.forename: Stefanie
authors.initials: S
authors.lastname: Kuhlmann
authors.forename: Simon
authors.initials: S
authors.lastname: Kollnberger
authors.forename: Andreas
authors.initials: A
authors.lastname: Thiel
authors.forename: Paul
authors.initials: P
authors.lastname: Bowness
authors.forename: Joachim
authors.initials: J
authors.lastname: Sieper
mesh_terms: D016918:Arthritis, Reactive / Q000209:etiology / Q000276:immunology*; D001704:Biopolymers; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D002469:Cell Separation / Q000379:methods; D002690:Chlamydia Infections / Q000150:complications / Q000276:immunology*; D002692:Chlamydia trachomatis / Q000276:immunology*; D003713:Dendritic Cells / Q000276:immunology / Q000382:microbiology; D018984:Epitopes, T-Lymphocyte / Q000276:immunology*; D005434:Flow Cytometry; D015796:HLA-B27 Antigen / Q000737:chemistry / Q000276:immunology*; D004854:Herpesvirus 4, Human / Q000276:immunology; D006801:Humans; D018189:Immunomagnetic Separation; D018716:Molecular Mimicry; D000091402:Prohibitins; D017510:Protein Folding; D011994:Recombinant Proteins / Q000737:chemistry; D013167:Spondylitis, Ankylosing / Q000209:etiology; D013582:Synovial Fluid / Q000276:immunology; D016176:T-Lymphocyte Subsets / Q000276:immunology*
chemical_list: D001704:Biopolymers; D018984:Epitopes, T-Lymphocyte; D015796:HLA-B27 Antigen; C000717331:PHB2 protein, human; D000091402:Prohibitins; D011994:Recombinant Proteins