Cargando…

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

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 boos...

Descripción completa

Detalles Bibliográficos
Autores principales: Son, Eric T., Paul-Heng, Moumita, Leong, Mario, Wang, Chuanmin, Hill, Alexandra E., Denkova, Martina, Purcell, Anthony W., Mifsud, Nicole A., Sharland, Alexandra F.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9792548/
https://www.ncbi.nlm.nih.gov/pubmed/36525346
http://dx.doi.org/10.1016/j.xpro.2022.101943
Descripción
Sumario: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)