Studying Organelle Dynamics in B Cells During Immune Synapse Formation

J Vis Exp. 2019 Jun 1:(148). doi: 10.3791/59621.

Abstract

Recognition of surface-tethered antigens by the B cell receptor (BCR) triggers the formation of an immune synapse (IS), where both signaling and antigen uptake are coordinated. IS formation involves dynamic actin remodeling accompanied by the polarized recruitment to the synaptic membrane of the centrosome and associated intracellular organelles such as lysosomes and the Golgi apparatus. Initial stages of actin remodeling allow B cells to increase their cell surface and maximize the quantity of antigen-BCR complexes gathered at the synapse. Under certain conditions, when B cells recognize antigens associated to rigid surfaces, this process is coupled to the local recruitment and secretion of lysosomes, which can facilitate antigen extraction. Uptaken antigens are internalized into specialized endo-lysosome compartments for processing into peptides, which are loaded onto major histocompatibility complex II (MHC-II) molecules for further presentation to T helper cells. Therefore, studying organelle dynamics associated with the formation of an IS is crucial to understanding how B cells are activated. In the present article we will discuss both imaging and a biochemical technique used to study changes in intracellular organelle positioning and cytoskeleton rearrangements that are associated with the formation of an IS in B cells.

Publication types

  • Research Support, Non-U.S. Gov't
  • Video-Audio Media

MeSH terms

  • Actins / metabolism
  • Animals
  • Antigen Presentation* / immunology
  • Antigens / metabolism
  • Antigens, Surface
  • B-Lymphocytes / cytology
  • B-Lymphocytes / immunology*
  • Cell Membrane / metabolism
  • Cell Polarity*
  • Centrosome / metabolism
  • Centrosome / physiology*
  • Cytoskeleton / metabolism
  • Histocompatibility Antigens Class II
  • Humans
  • Lysosomes / metabolism
  • Organelles / metabolism
  • Organelles / physiology*
  • Receptors, Antigen, B-Cell / immunology*
  • Synaptic Membranes / metabolism

Substances

  • Actins
  • Antigens
  • Antigens, Surface
  • Histocompatibility Antigens Class II
  • Receptors, Antigen, B-Cell