Bi-specific MHC heterodimers for characterization of cross-reactive T cells
Authors
Shen, Zu T.Brehm, Michael A.
Daniels, Keith A.
Sigalov, Alexander B.
Selin, Liisa K.
Welsh, Raymond M.
Stern, Lawrence J.
Student Authors
Zu T. ShenUMass Chan Affiliations
Department of Biochemistry and Molecular PharmacologyDepartment of Medicine, Division of Diabetes
Department of Pathology
Document Type
Journal ArticlePublication Date
2010-10-22Keywords
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 ProteinsImmunology and Infectious Disease
Life Sciences
Medicine and Health Sciences
Metadata
Show full item recordAbstract
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.Source
J Biol Chem. 2010 Oct 22;285(43):33144-53. Link to article on publisher's siteDOI
10.1074/jbc.M110.141051Permanent Link to this Item
http://hdl.handle.net/20.500.14038/33144PubMed ID
20729210Related Resources
Link to article in PubMedae974a485f413a2113503eed53cd6c53
10.1074/jbc.M110.141051