αβ T cell receptor interactions with syngeneic and allogeneic ligands: Affinity measurements and crystallization

K. Christopher Garcia, Michelle D. Tallquist, Larry R. Pease, Anders Brunmark, Christopher A. Scott, Massimo Degano, Enrico A. Stura, Per A. Peterson, Ian A. Wilson, Luc Teyton

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127 Scopus citations

Abstract

Cellular immunity is mediated by the interaction of an αβ T cell receptor (TCR) with a peptide presented within the context of a major histocompatibility complex (MHC) molecule. Alloreactive T cells have αβ TCRs that can recognize both self- and foreign peptide-MHC (pMHC) complexes, implying that the TCR has significant complementarity with different pMHC. To characterize the molecular basis for alloreactive TCR recognition of pMHC, we have produced a soluble, recombinant form of an alloreactive αβ T cell receptor in Drosophila melanogaster cells. This recombinant TCR, 2C, is expressed as a correctly paired αβ heterodimer, with the chains covalently connected via a disulfide bond in the C-terminal region. The native conformation of the 2C TCR was probed by surface plasmon resonance (SPR) analysis by using conformation-specific monoclonal antibodies, as well as syngeneic and allogeneic pMHC ligands. The 2C interaction with H-2Kb-dEV8, H-2Kbm3-dEV8, H-2Kb-SIYR, and H-2L(d)-p2Ca spans a range of affinities from K(d) = 10-4 to 10-6 M for the syngeneic (H-2Kb) and allogeneic (H- 2Kbm3, H-2L(d)) ligands. In general, the syngeneic ligands bind with weaker affinities than the allogeneic ligands, consistent with current threshold models of thymic selection and T cell activation. Crystallization of the 2C TCR required proteolytic trimming of the C-terminal residues of the α and chains. X-ray quality crystals of complexes of 2C with H-2Kb-dEV8, H- 2Kbm3-dEV8 and H-2Kb-SIYR have been grown.

Original languageEnglish (US)
Pages (from-to)13838-13843
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume94
Issue number25
DOIs
StatePublished - Dec 9 1997

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    Garcia, K. C., Tallquist, M. D., Pease, L. R., Brunmark, A., Scott, C. A., Degano, M., Stura, E. A., Peterson, P. A., Wilson, I. A., & Teyton, L. (1997). αβ T cell receptor interactions with syngeneic and allogeneic ligands: Affinity measurements and crystallization. Proceedings of the National Academy of Sciences of the United States of America, 94(25), 13838-13843. https://doi.org/10.1073/pnas.94.25.13838