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DNA-Based Probes for Measuring Mechanical Forces in Cell-Cell Contacts: Application to B Cell Antigen Extraction from Immune Synapses.
Spillane, Katelyn M; Tolar, Pavel.
Afiliação
  • Spillane KM; Immune Receptor Activation Laboratory, The Francis Crick Institute, 1 Midland Road, London, NW1 1AT, UK.
  • Tolar P; Division of Immunology and Inflammation, Department of Medicine, Imperial College London, London, UK.
Methods Mol Biol ; 1707: 69-80, 2018.
Article em En | MEDLINE | ID: mdl-29388100
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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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Estresse Mecânico / Linfócitos B / Receptores de Antígenos de Linfócitos B / Comunicação Celular / Sinapses Imunológicas / Antígenos Limite: Animals / Humans Idioma: En Revista: Methods Mol Biol Assunto da revista: BIOLOGIA MOLECULAR Ano de publicação: 2018 Tipo de documento: Article País de afiliação: Reino Unido

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Estresse Mecânico / Linfócitos B / Receptores de Antígenos de Linfócitos B / Comunicação Celular / Sinapses Imunológicas / Antígenos Limite: Animals / Humans Idioma: En Revista: Methods Mol Biol Assunto da revista: BIOLOGIA MOLECULAR Ano de publicação: 2018 Tipo de documento: Article País de afiliação: Reino Unido