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.

Journal article

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.

Authors: Toshitani, K; Braud, V; Browning, M; Murray, N; Mcmichael, A

• Publication status: Published
• Journal: Proceedings of the National Academy of Sciences of the United States of America
• Volume: 93
• Issue: 1
• Pages: 236-240
• Publication date: 1996-01-01
• DOI: 10.1073/pnas.93.1.236
• EISSN: 1091-6490
• ISSN: 0027-8424
• Language: English
• Keywords: T-Lymphocytes, Cytotoxic; Humans; Lymphocyte Cooperation; Signal Transduction; Antigen-Presenting Cells; Cell Line; Base Sequence; Amino Acid Sequence; Molecular Sequence Data; beta 2-Microglobulin; Recombinant Fusion Proteins; DNA Primers; Oligopeptides; Transfection; HLA-A2 Antigen