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Covalent Assembly of a Soluble T Cell Receptor-peptide-major Histocompatibility Class I Complex

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Specialty Science
Date 1996 Jul 9
PMID 8692966
Citations 6
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Abstract

We used stepwise photochemical cross-linking for specifically assembling soluble and covalent complexes made of a T-cell antigen receptor (TCR) and a class I molecule of the major histocompatibility complex (MHC) bound to an antigenic peptide. For that purpose, we have produced in myeloma cells a single-chain Fv construct of a TCR specific for a photoreactive H-2Kd-peptide complex. Photochemical cross-linking of this TCR single-chain Fv with a soluble form of the photoreactive H-2Kd-peptide ligand resulted in the formation of a ternary covalent complex. We have characterized the soluble ternary complex and showed that it reacted with antibodies specific for epitopes located either on the native TCR or on the Kd molecules. By preventing the fast dissociation kinetics observed with most T cell receptors, this approach provides a means of preparing soluble TCR-peptide-MHC complexes on large-scale levels.

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References
1.
Gunning P, Leavitt J, MUSCAT G, Ng S, Kedes L . A human beta-actin expression vector system directs high-level accumulation of antisense transcripts. Proc Natl Acad Sci U S A. 1987; 84(14):4831-5. PMC: 305199. DOI: 10.1073/pnas.84.14.4831. View

2.
Abastado J, Lone Y, Casrouge A, Boulot G, Kourilsky P . Dimerization of soluble major histocompatibility complex-peptide complexes is sufficient for activation of T cell hybridoma and induction of unresponsiveness. J Exp Med. 1995; 182(2):439-47. PMC: 2192121. DOI: 10.1084/jem.182.2.439. View

3.
Kubo R, Born W, Kappler J, Marrack P, Pigeon M . Characterization of a monoclonal antibody which detects all murine alpha beta T cell receptors. J Immunol. 1989; 142(8):2736-42. View

4.
White J, Blackman M, Bill J, Kappler J, Marrack P, Gold D . Two better cell lines for making hybridomas expressing specific T cell receptors. J Immunol. 1989; 143(6):1822-5. View

5.
Gregoire C, Rebai N, Schweisguth F, Necker A, Mazza G, Auphan N . Engineered secreted T-cell receptor alpha beta heterodimers. Proc Natl Acad Sci U S A. 1991; 88(18):8077-81. PMC: 52449. DOI: 10.1073/pnas.88.18.8077. View