Quantitating MHC class II trafficking in primary dendritic cells using imaging flow cytometry

Journal of Immunological Methods
Cassandra M HenniesEdith Janssen

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.

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Citations

Related Concepts

Immune-Associated Antigens
Veiled Cells
Flow Cytometry
Lymphocyte Activation
CD4 Positive T Lymphocytes
Image Cytometry
Dendritic Cells
Flow Cytometry
Histocompatibility Antigens Class II
Laboratory mice

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