Soluble, High-Affinity Dimers of T-Cell Receptors and Class II Major Histocompatibility Complexes: Biochemical Probes for Analysis and Modulation of Immune Responses

• Published in: Cellular immunology 1999-03, Vol.192 (2), p.175-184
• Main Authors: Lebowitz, Michael S., O'Herrin, Sean M., Hamad, Abdel-Rahim A., Fahmy, Tarek, Marguet, Didier, Barnes, Nicholas C., Pardoll, Drew, Bieler, Joan G., Schneck, Jonathan P.
• Format: Article
• Language: English
• Subjects: Base Sequence; Dimerization; Histocompatibility Antigens Class II - chemistry; Histocompatibility Antigens Class II - metabolism; Humans; Hybridomas - immunology; Immunoglobulin G - metabolism; Lymphocyte Activation; Molecular Sequence Data; Receptors, Antigen, T-Cell - chemistry; Receptors, Antigen, T-Cell - metabolism; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - metabolism
• ISSN: 0008-8749; 1090-2163
• DOI: 10.1006/cimm.1999.1441

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 chimeric 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 MHC (MCCI-Ek2Ig) 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.MCCI-Ek2Ig 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 modulatingin vivocellular responses.