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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.

Type

Journal article

Journal

Proc Natl Acad Sci U S A

Publication Date

09/01/1996

Volume

93

Pages

236 - 240

Keywords

Amino Acid Sequence, Antigen-Presenting Cells, Base Sequence, Cell Line, DNA Primers, HLA-A2 Antigen, Humans, Lymphocyte Cooperation, Molecular Sequence Data, Oligopeptides, Recombinant Fusion Proteins, Signal Transduction, T-Lymphocytes, Cytotoxic, Transfection, beta 2-Microglobulin