Imaging Membrane Curvature inside a FcεRI-Centric Synapse in RBL-2H3 Cells Using TIRF Microscopy with Polarized Excitation.

Brown Brown,Bendesky Bendesky,Machado Machado,Hagen Hagen,Spendier Spendier

doi:10.3390/jimaging5070063

Abstract

Total internal reflection fluorescence microscopy with polarized excitation (P-TIRF) can be used to image nanoscale curvature phenomena in live cells. We used P-TIRF to visualize rat basophilic leukemia cells (RBL-2H3 cells) primed with fluorescent anti-dinitrophenyl (anti-DNP) immunoglobulin E (IgE) coming into contact with a supported lipid bilayer containing mobile, monovalent DNP, modeling an immunological synapse. The spatial relationship of the IgE-bound high affinity IgE receptor (FcεRI) to the ratio image of P-polarized excitation and S-polarized excitation was analyzed. These studies help correlate the dynamics of cell surface molecules with the mechanical properties of the plasma membrane during synapse formation.

Highlights

The ability of eukaryotic cells to communicate with each other is important for numerous biological processes, including cell growth, motility, and immune function
In this cell-cell conjugate CD3 and γδ T-cell receptors are strongly polarized toward the mast cell contact site resulting in an immunological synapse formation [4]
These findings suggests that mast cells, and possibly basophils, can serve as nonconventional antigen-presenting cells and may play a larger role in signaling between physically contacting cells

Summary

Introduction

The ability of eukaryotic cells to communicate with each other is important for numerous biological processes, including cell growth, motility, and immune function This communication can occur through soluble factors (cytokines, interleukins, etc.), or by physical cell–cell contact. Mast cells can engage γδ T cells to protect against viral infections by forming mast cell- γδ T cell conjugates In this cell-cell conjugate CD3 and γδ T-cell receptors are strongly polarized toward the mast cell contact site resulting in an immunological synapse formation [4]. These findings suggests that mast cells, and possibly basophils, can serve as nonconventional antigen-presenting cells and may play a larger role in signaling between physically contacting cells

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Results

Conclusion