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MHC II-EGFP Knock-in Mouse Model.

Jan Pačes1, Valéria Grobárová1, Zdeněk Zadražil1

  • 1Laboratory of Cell Immunology, Department of Cell Biology, Faculty of Science, Charles University, Prague, Czech Republic.

Current Protocols
|November 7, 2023
PubMed
Summary

A novel MHC II-EGFP knock-in mouse model allows in vivo visualization of antigen-presenting cells (APCs). This tool aids in studying immune cell dynamics, localization, and MHC II gene regulation for quantitative immunology research.

Keywords:
APCsMHC II-EGFP knock-in mouselight-sheet fluorescence microscopylymphoid organsquantitative immunology

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Area of Science:

  • Immunology
  • Molecular Biology
  • Microscopy

Background:

  • Visualizing and tracking antigen-presenting cells (APCs) is crucial for understanding immune responses.
  • Existing methods may lack the precision needed for detailed analysis of APCs and their interactions.
  • MHC class II (MHC II) molecules are central to adaptive immunity, presenting antigens to CD4+ T cells.

Purpose of the Study:

  • To introduce and characterize a new MHC II-EGFP knock-in mouse model for visualizing MHC-II-expressing cells in vivo.
  • To demonstrate the utility of this model for identifying APCs, analyzing tissues, and studying MHC II gene regulation.
  • To provide a tool for quantitative and systems immunology research.

Main Methods:

  • Generation of a knock-in mouse model expressing enhanced green fluorescent protein (EGFP) fused to MHC class II under the MHC II beta chain promoter.
  • Utilizing fluorescence microscopy techniques, including light-sheet fluorescence microscopy, for visualization and quantification of APCs.
  • Correlating MHC II expression (MHC II-EGFP) with surface fraction via antibody detection for gene regulation studies.

Main Results:

  • The MHC II-EGFP model enables precise in vivo identification and tracking of various APC populations (dendritic cells, B cells, macrophages, ILC3s).
  • EGFP fluorescence is retained after fixation, facilitating immunofluorescence studies and unbiased histological characterization.
  • The model allows for detailed analysis of APC distribution in tissues and organs, and investigation into MHC II gene expression regulation.

Conclusions:

  • The MHC II-EGFP knock-in mouse model is a valuable asset for quantitative and systems immunology.
  • It provides a powerful tool for precise cell identification, studying immune cell dynamics, and understanding MHC II gene regulation.
  • This model significantly advances research into immune responses and their underlying molecular mechanisms.