Bi-specific MHC heterodimers for characterization of cross-reactive T cells
Interdisciplinary Graduate Program
Department of Pathology; Department of Medicine, Division of Diabetes; Department of Biochemistry and Molecular Pharmacology
Medical Subject Headings
Animals; Antigens, Viral; CD8-Positive T-Lymphocytes; Cell Line; Cricetinae; Cross Reactions; H-2 Antigens; Humans; Lymphocytic choriomeningitis virus; Mice; Peptides; Protein Structure, Quaternary; Vaccinia virus; Viral Proteins
Immunology and Infectious Disease | Life Sciences | Medicine and Health Sciences
T cell cross-reactivity describes the phenomenon whereby a single T cell can recognize two or more different peptide antigens presented in complex with MHC proteins. Cross-reactive T cells have previously been characterized at the population level by cytokine secretion and MHC tetramer staining assays, but single-cell analysis is difficult or impossible using these methods. In this study, we describe development of a novel peptide-MHC heterodimer specific for cross-reactive T cells. MHC-peptide monomers were independently conjugated to hydrazide or aldehyde-containing cross-linkers using thiol-maleimide coupling at cysteine residues introduced into recombinant MHC heavy chain proteins. Hydrazone formation provided bi-specific MHC heterodimers carrying two different peptides. Using this approach we prepared heterodimers of the murine class I MHC protein H-2K(b) carrying peptides from lymphocytic choriomeningitis virus and vaccinia virus, and used these to identify cross-reactive CD8+ T cells recognizing both lymphocytic choriomeningitis virus and vaccinia virus antigens. A similar strategy could be used to develop reagents to analyze cross-reactive T cell responses in humans.
Rights and Permissions
J Biol Chem. 2010 Oct 22;285(43):33144-53. Link to article on publisher's site
Shen, Zu T.; Brehm, Michael A.; Daniels, Keith A.; Sigalov, Alexander B.; Selin, Liisa K.; Welsh, Raymond M.; and Stern, Lawrence J., "Bi-specific MHC heterodimers for characterization of cross-reactive T cells" (2010). GSBS Student Publications. Paper 1685.