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Analysis of Dendritic Cell Function Using Clec9A-DTR Transgenic Mice.

Piotr Tetlak1, Christiane Ruedl2

  • 1School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, Singapore, 637551, Singapore.

Methods in Molecular Biology (Clifton, N.J.)
|May 5, 2016
PubMed
Summary

The Clec9A-diphtheria toxin receptor (DTR) mouse model enables study of CD8(+) and CD103(+) dendritic cells (DCs) in disease. Protocols for DC isolation, depletion, and analysis are provided, with cerebral malaria as a functional example.

Keywords:
Clec9A/DNGR-1Cross-primingDendritic cell (DC) subsetsDiphtheria toxin receptor (DTR) transgenic miceInterferon-γ experimental cerebral malaria (ΕCM)

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Area of Science:

  • Immunology
  • Cell Biology
  • Infectious Diseases

Background:

  • CD8(+) dendritic cells (DCs) in lymphoid organs and CD103(+) DCs in nonlymphoid tissues play crucial roles in immune responses.
  • Understanding their specific functions in infectious diseases and inflammation requires effective experimental models.

Purpose of the Study:

  • To describe protocols for utilizing the Clec9A-diphtheria toxin receptor (DTR) transgenic mouse model.
  • To detail methods for isolating, depleting, and analyzing CD8(+) and CD103(+) DCs.
  • To demonstrate the application of this model in studying DC function during experimental cerebral malaria.

Main Methods:

  • Generation and utilization of Clec9A-DTR transgenic mice.
  • Isolation of CD8(+) and CD103(+) dendritic cells (DCs).
  • In vivo depletion strategies for targeted DC populations.
  • Multi-color flow cytometry for DC characterization.
  • Establishment of an experimental cerebral malaria model for functional assessment.

Main Results:

  • The Clec9A-DTR system allows for robust investigation of CD8(+) and CD103(+) DC functions.
  • Protocols for DC isolation, depletion, and flow cytometry analysis are established.
  • The model is effective for studying DC roles in the context of experimental cerebral malaria.
  • Advantages and limitations of the Clec9A-DTR system are discussed.

Conclusions:

  • The Clec9A-DTR mouse model is a valuable tool for dissecting the roles of specific dendritic cell subsets in immunity and disease.
  • The provided protocols facilitate the use of this model in immunological research.
  • Further studies can leverage this system to explore DC-mediated mechanisms in various inflammatory and infectious conditions.