Screening self-peptides for recognition by mouse alloreactive CD8+ T cells using direct ex vivo multimer staining

Autor: Eric T. Son, Moumita Paul-Heng, Mario Leong, Chuanmin Wang, Alexandra E. Hill, Martina Denkova, Anthony W. Purcell, Nicole A. Mifsud, Alexandra F. Sharland
Jazyk: angličtina
Rok vydání: 2023
Předmět:
Zdroj: STAR Protocols, Vol 4, Iss 1, Pp 101943- (2023)
Druh dokumentu: article
ISSN: 2666-1667
DOI: 10.1016/j.xpro.2022.101943
Popis: Summary: Here, we present a protocol to identify immunogenic self-peptide/allogeneic major histocompatibility complex (MHC) epitopes. We describe the generation of enriched alloreactive CD8+ T cells by priming mice with a skin graft expressing the allogeneic MHC class I molecule of interest, followed by boosting with a liver-specific AAV vector encoding the heavy chain of that donor MHC allomorph. We then use a peptide-exchange approach to assemble a range of peptide-MHC (pMHC) multimers for measuring recognition of the various epitopes by these alloreactive T cells.For complete details on the use and execution of this protocol, please refer to Son et al. (2021).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
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