Soluble, high-affinity dimers of T-cell receptors and class II major histocompatibility complexes: Biochemical probes for analysis and modulation of immune responses

Michael S. Lebowitz, Sean M. O'Herrin, Abdel Rahim A. Hamad, Tarek Fahmy, Didier Marguet, Nicholas C. Barnes, Drew Pardoll, Joan G. Bieler, Jonathan P. Schneck

Research output: Contribution to journalArticlepeer-review

Abstract

T cell receptors (TCR) and major histocompatibility complex (MHC) molecules are integral membrane proteins that have central roles in cell- mediated immune recognition. Therefore, soluble analogs of these molecules would be useful for analyzing and possibly modulating antigen-specific immune responses. However, due to the intrinsic low-affinity and inherent solubility problems, it has been difficult to produce soluble high-affinity analogs of TCR and class II MHC molecules. This report describes a general approach which solves this intrinsic low-affinity by constructing soluble divalent analogs using IgG as a molecular scaffold. The divalent nature of the complexes increases the avidity of the chimerio molecules for cognate ligands. The generality of this approach was studied by making soluble divalent analogs of two different classes of proteins, a TCR (2C TCR2Ig) and a class II IHC ((MCC)I-E(k)2Ig) molecule. Direct flow cytometry assays demonstrate that the divalent 2C TCR2Ig chimera retained the specificity of the native 2C TCR, while displaying increased avidity for cognate peptide/MHC ligands, resulting in a high-affinity probe capable of detecting interactions that heretofore have only been detected using surface plasmon resonance. TCR2IgG was also used in immunofluorescence studies to show ER localization of intracellular peptide-MHC complexes after peptide feeding. (MCC)I-E(k)2Ig chimeras were able to both stain and activate an MCC-specific T cell hybridoma. Construction and expression of these two diverse heterodimers demonstrate the generality of this approach. Furthermore, the increased avidity of these soluble divalent proteins makes these chimeric molecules potentially useful in clinical settings for probing and modulating in vivo cellular responses.

Original languageEnglish (US)
Pages (from-to)175-184
Number of pages10
JournalCellular Immunology
Volume192
Issue number2
DOIs
StatePublished - Mar 15 1999

ASJC Scopus subject areas

  • Immunology

Fingerprint Dive into the research topics of 'Soluble, high-affinity dimers of T-cell receptors and class II major histocompatibility complexes: Biochemical probes for analysis and modulation of immune responses'. Together they form a unique fingerprint.

Cite this