A three-domain T cell receptor is biologically active and specifically stains cell surface MHC/peptide complexes

• Published in: The Journal of immunology (1950) Vol. 158; no. 5; pp. 2218 - 2227
• Main Authors: Plaksin, D, Polakova, K, McPhie, P, Margulies, DH
• Format: Journal Article
• Language: English
• Published: United States Am Assoc Immnol 01-03-1997
• Subjects: AIDS/HIV; Amino Acid Sequence; Animals; Antibodies, Monoclonal - chemistry; Antibody Specificity; Antigen-Antibody Reactions; Base Sequence; Circular Dichroism; Epitopes - chemistry; Epitopes - immunology; H-2 Antigens - chemistry; Hybridomas - immunology; Immunoglobulins - chemistry; Lymphocyte Activation; Mice; Molecular Sequence Data; Peptides - chemistry; Peptides - immunology; Peptides - physiology; Protein Binding - immunology; Protein Structure, Secondary; Protein Structure, Tertiary; Receptors, Antigen, T-Cell - biosynthesis; Receptors, Antigen, T-Cell - chemistry; Receptors, Antigen, T-Cell - immunology; Staining and Labeling; T-Lymphocytes - immunology
• ISSN: 0022-1767; 1550-6606
• DOI: 10.4049/jimmunol.158.5.2218

We have expressed in bacteria a single-chain T cell receptor (scTCR) with specificity for an HIV gp120-derived peptide bound to the murine MHC-I molecule, H-2Dd. This scTCR consists of V alpha covalently linked to the VbetaCbeta domains that was solubilized, refolded, and purified in high yield. Specific binding of the scTCR to MHC/peptide complexes was demonstrated by surface plasmon resonance, with a Kd of 2 to 8 x 10(-6) M. This scTCR specifically inhibited T cell activation, and stained cell surface MHC/peptide complexes as measured by cytofluorimetry. The preservation of binding specificity by such a three-domain scTCR suggests that this structure is sufficient for specific MHC/peptide recognition and that this strategy will be of general use as applied to other TCR.