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Updated: Apr 12, 2026
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Published on: October 29, 2018
Quantitating MHC class II trafficking in primary dendritic cells using imaging flow cytometry
Cassandra M Hennies1, Maria A Lehn1, Edith M Janssen1
1Division of Immunobiology, Cincinnati Children's Hospital Medical Center and the University of Cincinnati College of Medicine, Cincinnati, OH, USA.
Insights
This study introduces a new imaging flow cytometry assay to visualize MHC class II (MHCII) trafficking in dendritic cells (DCs). This tool helps uncover molecular mechanisms regulating MHCII levels and CD4(+) T cell activation.
Area of Science:
- Immunology
- Cell Biology
Background:
- Dendritic cells (DCs) present antigenic peptides via MHC class II (MHCII) molecules, initiating CD4(+) T cell activation.
- Surface MHCII-peptide complex levels critically influence CD4(+) T cell response magnitude and phenotype.
- The molecular regulation of MHCII surface expression, including trafficking dynamics, remains incompletely understood.
Purpose of the Study:
- To develop and validate a novel multispectral imaging flow cytometry assay.
- To visualize and analyze MHCII trafficking dynamics in primary dendritic cells.
- To provide a tool for dissecting molecular mechanisms governing MHCII homeostasis.
Main Methods:
- Multispectral imaging flow cytometry.
- Visualization of MHCII trafficking in primary mouse and human dendritic cells.
- Assay development for dissecting molecular mechanisms of MHCII regulation.
Main Results:
- Successfully developed a multispectral imaging flow cytometry assay for visualizing MHCII trafficking.
- Demonstrated the assay's utility in primary mouse and human dendritic cells.
- Established a tool to investigate the molecular basis of MHCII surface expression regulation.
Conclusions:
- The developed assay is a valuable tool for studying MHCII trafficking and homeostasis in dendritic cells.
- This method facilitates the dissection of molecular mechanisms controlling MHCII surface presentation.
- Understanding MHCII regulation is crucial for modulating CD4(+) T cell responses.
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.
