Chapter title |
DNA-Based Probes for Measuring Mechanical Forces in Cell-Cell Contacts: Application to B Cell Antigen Extraction from Immune Synapses
|
---|---|
Chapter number | 5 |
Book title |
B Cell Receptor Signaling
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-7474-0_5 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7473-3, 978-1-4939-7474-0
|
Authors |
Katelyn M. Spillane, Pavel Tolar |
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. |
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