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Bi‐specific MHC Heterodimers for Characterization of Cross‐reactive T Cells
Author(s) -
Shen Zu Ting,
Brehm Michael A,
Daniels Keith A,
Sig Alexander B,
Selin Liisa K,
Welsh Raymond M,
Stern Lawrence
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.lb106
Subject(s) - lymphocytic choriomeningitis , major histocompatibility complex , mhc class i , t cell , mhc restriction , cytotoxic t cell , antigen , biology , cd8 , microbiology and biotechnology , chemistry , biochemistry , in vitro , immune system , immunology
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.