title: Identification of HLA-A2 restricted T-cell epitopes within the conserved region of the immunoglobulin G heavy-chain in patients with multiple myeloma.
abstract: OBJECTIVE
  The aim of this study is the identification of HLA-A2 restricted T-cell epitopes in the conserved region of the immunoglobulin-G-heavy-chain (IgGH) that can be used for immunotherapy in multiple myeloma (MM) patients.
  
  
  METHODS
  After the IgGH gene sequence was scanned for HLA-A2 restricted T-cell epitopes with a high binding affinity to the MHC-I-complex, promising nona-peptides were synthesized. Peptide specific CD8+ T-cells were generated from peripheral blood mononuclear cells (PBMC) of healthy donors (HD) and patients with MM using peptide pulsed dendritic cells (DC) in vitro. The activation and cytotoxicity of CD8+ T-cells was analyzed by IFN-alpha ELISpot-assay and 51Chromium release-assay. HLA-A2 restriction was proven by blocking T-cell activation with anti-HLA-A2 antibodies.
  
  
  RESULTS
  Two HLA-A2 restricted T-cell epitopes-TLVTVSSAS derived from the IgGH-framework-region 4 (FR4) and LMISRTPEV from the constant region (CR)-induced expansion of specific CD8+ T-cells from PBMC in two of three (TLVTVSSAS) and one of three (LMISRTPEV) HD respectively. Specific T-cells were induced from PBMC in two of six (TLVTVSSAS) and eight of 19 (LMISRTPEV) patients with MM. Specific CD8+ T-cells also lysed peptide-pulsed target cells in 51Chromium release-assay. LMISRTPEV specific CD8+ T-cells from MM patients lysed specifically the HLA-A2+ IgG myeloma cell line XG-6.
  
  
  CONCLUSIONS
  We identified two HLA-A2 restricted T-cell epitopes-TLVTVSSAS and LMISRTPEV--which can yield an expansion of CD8+ T-cells with the ability to kill peptide-loaded target cells and HLA-A2+ IgG+ myeloma cells. We conclude that TLVTVSSAS and LMISRTPEV could be T-cell epitopes for immunotherapy in MM patients.
authors: Department of internal medicine V University of HeidelbergHeidelberg,DE.
authors: Sebastian
authors: S
authors: Belle
authors: Department of internal medicine V University of HeidelbergHeidelberg,DE.
authors: Fang
authors: F
authors: Han
authors: Immunopathologie des maladies tumorales et autoimmunes INSERM : U475IFR76Institut de recherche en biothérapieUniversité Montpellier ICentre de Recherche Inserm 99, Rue Puech Villa 34197 MONTPELLIER CEDEX 5,FR.|IRB, Institut de recherche en biothérapie CHRU MontpellierUniversité Montpellier IHopital Saint-Eloi 34000 Montpellier,FR.
authors: Maud
authors: M
authors: Condomines
authors: Department of internal medicine V University of HeidelbergHeidelberg,DE.
authors: Olaf
authors: O
authors: Christensen
authors: Department of internal medicine V University of HeidelbergHeidelberg,DE.
authors: Mathias
authors: M
authors: Witzens-Harig
authors: Department of internal medicine V University of HeidelbergHeidelberg,DE.
authors: Bernd
authors: B
authors: Kasper
authors: Department of transplantation immunology University of HeidelbergFR.
authors: Christian
authors: C
authors: Kleist
authors: Department of transplantation immunology University of HeidelbergFR.
authors: Peter
authors: P
authors: Terness
authors: Department of internal medicine V University of HeidelbergHeidelberg,DE.
authors: Marion
authors: M
authors: Moos
authors: Department of internal medicine V University of HeidelbergHeidelberg,DE.
authors: Friedrich
authors: F
authors: Cremer
authors: Department of internal medicine V University of HeidelbergHeidelberg,DE.
authors: Dirk
authors: D
authors: Hose
authors: Department of internal medicine V University of HeidelbergHeidelberg,DE.
authors: Anthony D
authors: AD
authors: Ho
authors: Department of internal medicine V University of HeidelbergHeidelberg,DE.
authors: Hartmut
authors: H
authors: Goldschmidt
authors: Immunopathologie des maladies tumorales et autoimmunes INSERM : U475IFR76Institut de recherche en biothérapieUniversité Montpellier ICentre de Recherche Inserm 99, Rue Puech Villa 34197 MONTPELLIER CEDEX 5,FR.|IRB, Institut de recherche en biothérapie CHRU MontpellierUniversité Montpellier IHopital Saint-Eloi 34000 Montpellier,FR.
authors: Bernard
authors: B
authors: Klein
authors: Department of internal medicine V University of HeidelbergHeidelberg,DE.
authors: Michael
authors: M
authors: Hundemer
mesh_terms: D000595:Amino Acid Sequence; D017951:Antigen Presentation; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D003602:Cytotoxicity, Immunologic; D003713:Dendritic Cells / Q000276:immunology; D018984:Epitopes, T-Lymphocyte / Q000276:immunology*; D015789:HLA-A2 Antigen / Q000276:immunology*; D006801:Humans; D007127:Immunoglobulin Constant Regions; D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains / Q000276:immunology*; D007167:Immunotherapy; D009101:Multiple Myeloma / Q000276:immunology* / Q000628:therapy
chemical_list: D018984:Epitopes, T-Lymphocyte; D015789:HLA-A2 Antigen; D007127:Immunoglobulin Constant Regions; D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains