title: Evaluation of viral interference with MHC class I-restricted antigen processing and presentation using a flow cytometry-based approach.
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.
authors: Department of Medical Microbiology, University Medical Center Utrecht, Utrecht, The Netherlands.
authors: Daniëlle
authors: D
authors: Horst
authors: Department of Medical Microbiology, University Medical Center Utrecht, Utrecht, The Netherlands.
authors: Maaike E
authors: ME
authors: Ressing
authors: Department of Immunohaematology and Blood Transfusion, Leiden University Medical Center, Leiden, The Netherlands.
authors: Arend
authors: A
authors: Mulder
authors: Department of Medical Microbiology, University Medical Center Utrecht, Utrecht, The Netherlands. e.wiertz@umcutrecht.nl.
authors: Emmanuel J H J
authors: EJHJ
authors: Wiertz
mesh_terms: D017951:Antigen Presentation; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology / Q000821:virology; D002460:Cell Line; D005434:Flow Cytometry / Q000379:methods*; D015395:Histocompatibility Antigens Class I / Q000276:immunology*; D016086:Lentivirus / Q000235:genetics / Q000502:physiology*; D013194:Staining and Labeling; D014162:Transfection
chemical_list: D015395:Histocompatibility Antigens Class I