Expression of a single-chain HLA class I molecule in a human cell line: presentation of exogenous peptide and processed antigen to cytotoxic T lymphocytes
| Autor: | Koji Toshitani, Veronique M. Braud, Nicholas Murray, M. J. Browning, Walter F. Bodmer, Andrew J. McMichael |
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| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
Recombinant Fusion Proteins
Lymphocyte Cooperation Molecular Sequence Data Antigen-Presenting Cells Human leukocyte antigen Biology Transfection Cell Line Antigen HLA-A2 Antigen medicine Humans Cytotoxic T cell Interferon gamma Amino Acid Sequence Antigen-presenting cell Peptide sequence DNA Primers Multidisciplinary Base Sequence Antigen processing Molecular biology beta 2-Microglobulin Oligopeptides Research Article Signal Transduction T-Lymphocytes Cytotoxic medicine.drug |
| DOI: | 10.1073/pnas.93.1.236 |
| Popis: | We have synthesized a recombinant gene encoding a single-chain HLA-A2/beta 2-microglobulin (beta 2m) molecule by linking beta 2m through its carboxyl terminus via a short peptide spacer to HLA-A2 (A*0201). This gene has been expressed in the beta 2m-deficient colorectal tumor cell line DLD-1. Transfection of this cell with the single-chain construct was associated with conformationally correct cell surface expression of a class I molecule of appropriate molecular mass. The single-chain HLA class I molecule presented either exogenously added peptide or (after interferon-gamma treatment) endogenously processed antigen to an influenza A matrix-specific, HLA-A2-restricted cytotoxic T-lymphocyte line. The need for interferon gamma for the processing and presentation of endogenous antigen suggests that DLD-1 has an antigen-processing defect that can be up-regulated, a feature that may be found in other carcinomas. Our data indicate that single-chain HLA class I constructs can form functional class I molecules capable of presenting endogenously processed antigens. Such molecules should be of use for functional studies, as well as providing potential anticancer immunotherapeutic agents or vaccines. |
| Databáze: | OpenAIRE |
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