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Imaging the Interactions Between B Cells and Antigen-Presenting Cells.

Jia C Wang1, Madison Bolger-Munro1, Michael R Gold2

  • 1Department of Microbiology & Immunology and the Life Sciences Institute, University of British Columbia, 2350 Health Sciences Mall, Vancouver, BC, V6T 1Z3, Canada.

Methods in Molecular Biology (Clifton, N.J.)
|February 2, 2018
PubMed
Summary
This summary is machine-generated.

Researchers developed a new method to study B cell activation using model antigens on Cos-7 cells. This approach allows detailed observation of immune synapse formation and B cell receptor signaling dynamics.

Keywords:
Antigen-presenting cellB cell receptorB cellsConfocal microscopyCytoskeletonImmune synapseSignal transduction

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Area of Science:

  • Immunology
  • Cell Biology
  • Biophysics

Background:

  • B cell activation is crucial for adaptive immunity, typically initiated by antigen presentation on antigen-presenting cells (APCs).
  • The B cell receptor (BCR) interaction with membrane-bound antigens triggers rapid cellular rearrangements, forming an immune synapse.
  • Previous models used artificial systems, limiting physiological relevance for B cell-APC interactions.

Purpose of the Study:

  • To establish a more physiologically relevant model for studying B cell-APC interactions.
  • To investigate B cell immune synapse formation, including cytoskeletal dynamics and BCR organization.
  • To analyze BCR-mediated antigen capture and signaling pathways.

Main Methods:

  • Expression of model transmembrane antigens (anti-Igκ antibody, hen egg lysozyme-fluorescent protein) on Cos-7 cells.
  • Transfection of B cells with siRNAs or fluorescent protein plasmids.
  • Utilizing fixed and live-cell fluorescence microscopy to visualize B cell-APC interactions.
  • Quantification of changes in BCR spatial organization and signaling upon APC contact.

Main Results:

  • Successfully generated Cos-7 cells presenting model antigens for B cell interaction studies.
  • Observed and quantified cytoskeletal reorganization, BCR microcluster coalescence, and antigen gathering.
  • Demonstrated the utility of this system for analyzing BCR signaling dynamics in response to antigen presentation.
  • Provided detailed protocols for replicating these experimental setups.

Conclusions:

  • The Cos-7 cell-based model provides a robust and adaptable system for studying B cell activation and immune synapse formation.
  • This methodology enables detailed investigation of the molecular mechanisms underlying B cell-APC interactions.
  • The developed protocols facilitate further research into B cell immunology and signaling pathways.