Multiplex MHC Class I Tetramer Combined with Intranuclear Staining by Mass Cytometry
Antigen-specific CD8+ T cells play a crucial role in the host protective immune response against viruses, tumors, and other diseases. Major histocompatibility complex (MHC) class I tetramers allow for a direct detection of such antigen-specific CD8+ T cells. Using mass cytometry together with multiplex MHC class I tetramer staining, we are able to screen more than 1000 different antigen candidates simultaneously across tissues in health and disease, while retaining the possibility to deliver an in-depth characterization of antigen-specific CD8+ T cells and associated phenotypes. Here we describe the method for a MHC class I tetramer multiplexing approach together with intracellular antibody staining for a parallel phenotypic cell characterization using mass cytometry in human specimens.
Key wordsMass cytometry MHC class I tetramer Antigen-specific T cell Multiplex tetramer staining
The authors thank all members of Evan Newell lab.
Competing interests: E.W.N. is a board director and shareholder of immunoSCAPE Pte.Ltd. M.F. is Director, Scientific Affairs and shareholder of immunoSCAPE Pte. Ltd. All other authors declare no competing financial interests.
- 1.Wong P, Pamer EG (2003) CD8 T cell responses to infectious pathogens. Annu Rev Immunol 21:29–70. https://doi.org/10.1146/annurev.immunol.21.120601.141114CrossRefPubMedGoogle Scholar
- 27.Becht E, Dutertre C-A, Kwok IWH, et al (2018) Evaluation of UMAP as an alternative to t-SNE for single-cell data. bioRxiv. https://doi.org/10.1101/298430