DNA-based probes for measuring mechanical forces in cell-cell contacts: Application to B cell antigen extraction from immune synapses
File(s)Spillane, Molecular Immunology, 2018 - 180226.pdf (332.63 KB)
Accepted version
Author(s)
Spillane, Katelyn M
Tolar, Pavel
Type
Journal Article
Abstract
The production of antibodies requires the expansion and selection of high-affinity B cell clones. This process is initiated by antigen uptake through the B cell receptor (BCR), which recognizes and binds antigen displayed on the surface of an antigen-presenting cell (APC). To acquire the antigen, B cells use myosin contractility to physically pull BCR-antigen clusters from the APC membrane. These mechanical forces influence association and dissociation rates of BCR-antigen bonds, resulting in affinity-dependent acquisition of antigen by B cells. Mechanical regulation of B cell antigen acquisition from APCs remains poorly understood, although the recent development of DNA-based force sensors has enabled the measurement of mechanical forces generated in B cell-APC contacts. In this chapter, we describe a protocol to design, synthesize, and purify DNA-based force sensors to measure B cell antigen extraction forces using fluorescence microscopy.
Date Issued
2018-02-01
Date Acceptance
2017-10-02
Citation
Methods in Molecular Biology, 2018, 1707, pp.69-80
ISSN
1940-6029
Publisher
Humana Press
Start Page
69
End Page
80
Journal / Book Title
Methods in Molecular Biology
Volume
1707
Copyright Statement
© 2018 Springer Science+Business Media, LLC. The final publication is available at Springer via https://dx.doi.org/10.1007/978-1-4939-7474-0_5
Identifier
https://www.ncbi.nlm.nih.gov/pubmed/29388100
Subjects
Antigen internalization
B cell receptor
B cells
Cell mechanics
DNA force sensor
DNA-protein conjugation
Fluorescence microscopy
Immune synapse
Animals
Antigens
B-Lymphocytes
Cell Communication
Humans
Immunological Synapses
Receptors, Antigen, B-Cell
Stress, Mechanical
Publication Status
Published
Coverage Spatial
United States
Date Publish Online
2018-02-01