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                <title xml:lang="en">Identification of a new HLA-A2-restricted T-cell epitope within HM1.24 as immunotherapy target for multiple myeloma.</title>
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                    <forename type="first">Maud</forename>
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                  <idno type="halauthorid">193046-0</idno>
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                  <idno type="halauthorid">194217-0</idno>
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                  <persName>
                    <forename type="first">Dirk</forename>
                    <surname>Hose</surname>
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                  <idno type="halauthorid">191308-0</idno>
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                  <persName>
                    <forename type="first">Marion</forename>
                    <surname>Moos</surname>
                  </persName>
                  <idno type="halauthorid">191304-0</idno>
                  <affiliation ref="#struct-29781"/>
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                <author role="aut">
                  <persName>
                    <forename type="first">Friedrich</forename>
                    <forename type="middle">W.</forename>
                    <surname>Cremer</surname>
                  </persName>
                  <idno type="halauthorid">191299-0</idno>
                  <affiliation ref="#struct-29781"/>
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                  <persName>
                    <forename type="first">Christian</forename>
                    <surname>Kleist</surname>
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                  <idno type="halauthorid">194218-0</idno>
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                  <idno type="halauthorid">194219-0</idno>
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                    <forename type="first">Bernard</forename>
                    <surname>Klein</surname>
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              <classCode scheme="mesh">Antigens, CD</classCode>
              <classCode scheme="mesh">Cell Line, Tumor</classCode>
              <classCode scheme="mesh">Multiple Myeloma</classCode>
              <classCode scheme="mesh">Neoplasm Proteins</classCode>
              <classCode scheme="mesh">Oligopeptides</classCode>
              <classCode scheme="mesh">Plasma Cells</classCode>
              <classCode scheme="mesh">T-Lymphocytes, Cy</classCode>
              <classCode scheme="mesh">Dendritic Cells</classCode>
              <classCode scheme="mesh">Epitopes, T-Lymphocyte</classCode>
              <classCode scheme="mesh">HLA-A2 Antigen</classCode>
              <classCode scheme="mesh">Humans</classCode>
              <classCode scheme="mesh">Immunotherapy</classCode>
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              <classCode scheme="mesh">Lymphocyte Activation</classCode>
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              <p>OBJECTIVE: The aim of this study was identification of human leukocyte antigen (HLA)-A2-restricted T-cell epitopes within the HM1.24 antigen as target for multiple myeloma (MM)-directed specific peptide-based immunotherapy. METHODS: The HM1.24 sequence was scanned for immunogenic peptides using the HLA-binding prediction software SYFPEITHI and BIMAS. Peripheral blood mononuclear cells from HLA-A2(+) healthy volunteers/blood donors (ND) were stimulated with autologous HM1.24-peptide-loaded dendritic cells, and expanded in vitro. Activation of T cells was assessed by ELISpot and cytotoxicity by (51)Chromium ((51)Cr)-release assays. T2-cells pulsed with irrelevant peptide, the HM1.24(-)/HLA-A2(+) breast carcinoma cell line MCF-7 and the HM1.24(+)/HLA-A2(-) myeloma cell line RPMI-8226 were used as controls. Expression of the HM1.24 gene (BST2) was assessed using purified plasma cells and Affymetrix-U133A+B microarrays. Frequency of peptide-specific CD8(+) T cells was detected using the flow-cytometric tetramer technique. RESULTS: Of eight nona-peptides with the highest probability of binding to HLA-A2, the HM1.24 aa22-30 peptide (LLLGIGILV) showed the most frequent activation of CD8(+) T cells in healthy volunteers (specific activation in 8 of 11 [73%] ND; compared with 5-19% for the 7 other HM1.24 peptides). Antigen recognition by the HM1.24 aa22-30-specific CD8(+) T cells was HLA-A2-restricted (ELISpot with HLA-A2-blocking antibodies: median, 15; range, 14-18 spots/well; isotype-control antibodies: median, 47; range, 44-48). HM1.24-aa22-30-specific CD8(+) T cells lysed HLA-A2(+) myeloma-derived cell lines ((51)Cr-release assay: XG-1 vs MCF-7, 91% vs 0%; U266 vs MCF-7, 38% vs 4.2%; IM-9 vs RPMI-8226, 22% vs 0%). Using the cross-reactive Neisseria meningitidis peptide LLSLGIGILV-specific CD8(+) T cells recognizing target cells loaded with the HM1.24 aa22-30 peptide (LLLGIGILV) as well as the myeloma-derived cell line U266 could be expanded from MM patients. The HM1.24 gene was expressed at comparable levels by plasma cells from 65 MM patients, 7 patients with monoclonal gammopathy of undetermined significance, and 7 ND. CONCLUSIONS: HM1.24 aa22-30 is a newly identified HLA-A2-restricted T-cell epitope that is processed and presented by major histocompatibility complex class I. Specifically activated CD8(+) T cells are able to lyse MM cell lines. We conclude that HM1.24 aa22-30 represents a suitable candidate target for a specific peptide-based immunotherapy of MM.</p>
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