title: Quantitating MHC class II trafficking in primary dendritic cells using imaging flow cytometry.
abstract: Presentation of antigenic peptides in MHC class II (MHCII) on dendritic cells (DCs) is the first step in the activation of antigen-specific CD4(+)T cells. The expression of surface MHCII-peptide complexes is tightly regulated as the frequency of MHCII-peptide complexes can affect the magnitude, as well as the phenotype of the ensuing CD4(+)T cell response. The surface MHCII-peptide levels are determined by the balance between expression of newly generated complexes, complex internalization, and their subsequent re-emergence or degradation. However, the molecular mechanisms that underpin these processes are still poorly understood. Here we describe a multispectral imaging flow cytometry assay to visualize MHCII trafficking that can be used as a tool to dissect the molecular mechanisms that regulate MHCII homeostasis in primary mouse and human DCs.
authors.affiliation: Division of Immunobiology, Cincinnati Children's Hospital Medical Center and the University of Cincinnati College of Medicine, Cincinnati, OH, USA.
authors.forename: Cassandra M
authors.initials: CM
authors.lastname: Hennies
authors.affiliation: Division of Immunobiology, Cincinnati Children's Hospital Medical Center and the University of Cincinnati College of Medicine, Cincinnati, OH, USA.
authors.forename: Maria A
authors.initials: MA
authors.lastname: Lehn
authors.affiliation: Division of Immunobiology, Cincinnati Children's Hospital Medical Center and the University of Cincinnati College of Medicine, Cincinnati, OH, USA. Electronic address: Edith.Janssen@cchmc.org.
authors.forename: Edith M
authors.initials: EM
authors.lastname: Janssen
mesh_terms: D000818:Animals; D001692:Biological Transport / Q000502:physiology*; D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism; D002460:Cell Line; D003713:Dendritic Cells / Q000378:metabolism*; D005434:Flow Cytometry / Q000379:methods; D000949:Histocompatibility Antigens Class II / Q000378:metabolism*; D006801:Humans; D019044:Image Cytometry / Q000379:methods; D008213:Lymphocyte Activation / Q000502:physiology; D051379:Mice
chemical_list: D000949:Histocompatibility Antigens Class II
keywords: Dendritic cells; Imaging; Internalization; MHC class II