Bi-specific MHC heterodimers for characterization of cross-reactive T cells.
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.
Shen
Zu T
ZT
From the Department of Pathology, Worcester, Massachusetts 01655.
Brehm
Michael A
MA
Division of Diabetes, Worcester, Massachusetts 01655.
Daniels
Keith A
KA
From the Department of Pathology, Worcester, Massachusetts 01655.
Sigalov
Alexander B
AB
From the Department of Pathology, Worcester, Massachusetts 01655.
Selin
Liisa K
LK
From the Department of Pathology, Worcester, Massachusetts 01655.
Welsh
Raymond M
RM
From the Department of Pathology, Worcester, Massachusetts 01655.
Stern
Lawrence J
LJ
From the Department of Pathology, Worcester, Massachusetts 01655; Department of Biochemistry & Molecular Pharmacology, University of Massachusetts Medical School, Worcester, Massachusetts 01655. Electronic address: lawrence.stern@umassmed.edu.
D000818:Animals; D000956:Antigens, Viral / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D002460:Cell Line; D006224:Cricetinae; D003429:Cross Reactions / Q000276:immunology; D006183:H-2 Antigens / Q000276:immunology*; D006801:Humans; D008217:Lymphocytic choriomeningitis virus / Q000276:immunology*; D051379:Mice; D010455:Peptides / Q000276:immunology*; D020836:Protein Structure, Quaternary; D014616:Vaccinia virus / Q000276:immunology*; D014764:Viral Proteins / Q000276:immunology*
D000956:Antigens, Viral; D006183:H-2 Antigens; C032876:H-2Kb protein, mouse; D010455:Peptides; D014764:Viral Proteins