Abstract
The peptide content of MHC class I molecules present at the cell surface is monitored by surveilling CD8+ cytotoxic T cells. In case of a viral infection, a proportion of the MHC class I molecules will carry peptides derived from viral proteins. This allows the CD8+ T cells to recognize and eliminate virus-infected cells. This highly sensitive detection system of the host is counteracted by viruses, which have acquired functions to downregulate cell surface expression of MHC class I molecules. In this chapter, we describe a flow cytometry-based method to identify viral gene product(s) responsible for evasion from MHC class I-restricted antigen presentation. To this end, cells are transiently transfected using polyethylenimine (PEI) as a transfection reagent, followed by cell surface staining with MHC class I-specific monoclonal antibodies. Once viral proteins responsible for MHC class I downregulation have been identified, their mechanism of action can be characterized. Identification and characterization of virus-encoded MHC class I inhibitors augments our understanding of virus–host interactions and often provides new insights into antigen processing and presentation pathways, including related cellular processes such as protein trafficking and degradation.
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Acknowledgements
This work was supported by the Dutch Cancer Foundation (grant UU 2005-3259 to D.H., M.R., and E.W.) and the Netherlands Scientific Organization (NWO Vidi 917.76.330.1 to M.R.).
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Horst, D., Ressing, M.E., Mulder, A., Wiertz, E.J.H.J. (2013). Evaluation of Viral Interference with MHC Class I-Restricted Antigen Processing and Presentation Using a Flow Cytometry-Based Approach. In: van Endert, P. (eds) Antigen Processing. Methods in Molecular Biology™, vol 960. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-218-6_10
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DOI: https://doi.org/10.1007/978-1-62703-218-6_10
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