The University of Southampton
University of Southampton Institutional Repository

Real-time activation profiles of a single T cell arrays following controlled interaction with antigen-presenting cells

Real-time activation profiles of a single T cell arrays following controlled interaction with antigen-presenting cells
Real-time activation profiles of a single T cell arrays following controlled interaction with antigen-presenting cells
The aim of this project was to develop a new microdevice to allow precise control of cellcell interactions, and to apply it to measure T cell signalling in arrays of single cells in realtime after stimulation by live antigen presenting cells.

The specific T cell responses which underlie effective immunotherapy of cancer are mediated via cell-cell interactions and the formation of immune synapses. However, methods to study these interactions are limited by lack of control over single cell pairing and analysis methods. This project describes a new approach to track the activation profile of cytotoxic T cells against tumour cells in a high-throughput manner.

A simple yet effective mechanism to pair the single T cells with antigen presenting cells was optimized to study cell-cell contact in a time-controlled manner. The cell-trap array is an open system consisting of thousands of microwells cast in an agarose block, which is biocompatible and permeable to nutrients allowing functional observations on viable cells to be carried out over a number of hours. T cells can be singularly isolated in the wells via passive sedimentation and size exclusion, achieving up to 90% occupancy.

Activation profiles of thousands of single CD8+ cells could be screened in the same field of view using calcium-chelating fluorophore dyes to measure the early stages of T cell signalling. Custom software was developed to process experimental data and allow rapid detection of individual cells with the strongest activation signals defined by several different criteria. This capability could be used in future to identify and characterise single T-cells that have especially high propensity to be activated by immunotherapy such as vaccination or checkpoint blockade, in preclinical animal models and biopsy samples from human cancer patients.
University of Southampton
Desalvo, Anna
45bbaca1-d004-4a5c-a9cd-3c58f5e2ee0d
Desalvo, Anna
45bbaca1-d004-4a5c-a9cd-3c58f5e2ee0d
Elliott, Timothy
16670fa8-c2f9-477a-91df-7c9e5b453e0e
Morgan, Hywel
de00d59f-a5a2-48c4-a99a-1d5dd7854174
James, Edward
7dc1afb7-d326-4050-89fc-1f4e2a1a19a4

Desalvo, Anna (2019) Real-time activation profiles of a single T cell arrays following controlled interaction with antigen-presenting cells. University of Southampton, Doctoral Thesis, 243pp.

Record type: Thesis (Doctoral)

Abstract

The aim of this project was to develop a new microdevice to allow precise control of cellcell interactions, and to apply it to measure T cell signalling in arrays of single cells in realtime after stimulation by live antigen presenting cells.

The specific T cell responses which underlie effective immunotherapy of cancer are mediated via cell-cell interactions and the formation of immune synapses. However, methods to study these interactions are limited by lack of control over single cell pairing and analysis methods. This project describes a new approach to track the activation profile of cytotoxic T cells against tumour cells in a high-throughput manner.

A simple yet effective mechanism to pair the single T cells with antigen presenting cells was optimized to study cell-cell contact in a time-controlled manner. The cell-trap array is an open system consisting of thousands of microwells cast in an agarose block, which is biocompatible and permeable to nutrients allowing functional observations on viable cells to be carried out over a number of hours. T cells can be singularly isolated in the wells via passive sedimentation and size exclusion, achieving up to 90% occupancy.

Activation profiles of thousands of single CD8+ cells could be screened in the same field of view using calcium-chelating fluorophore dyes to measure the early stages of T cell signalling. Custom software was developed to process experimental data and allow rapid detection of individual cells with the strongest activation signals defined by several different criteria. This capability could be used in future to identify and characterise single T-cells that have especially high propensity to be activated by immunotherapy such as vaccination or checkpoint blockade, in preclinical animal models and biopsy samples from human cancer patients.

Text
PhD Thesis Anna Desalvo - Version of Record
Available under License University of Southampton Thesis Licence.
Download (29MB)

More information

Published date: May 2019

Identifiers

Local EPrints ID: 437706
URI: http://eprints.soton.ac.uk/id/eprint/437706
PURE UUID: f4af73f5-f95b-4a0c-9489-11fa9b2218e5
ORCID for Anna Desalvo: ORCID iD orcid.org/0000-0003-1357-3222
ORCID for Timothy Elliott: ORCID iD orcid.org/0000-0003-1097-0222
ORCID for Hywel Morgan: ORCID iD orcid.org/0000-0003-4850-5676
ORCID for Edward James: ORCID iD orcid.org/0000-0001-8638-7928

Catalogue record

Date deposited: 12 Feb 2020 17:31
Last modified: 18 Feb 2021 17:07

Export record

Contributors

Author: Anna Desalvo ORCID iD
Thesis advisor: Timothy Elliott ORCID iD
Thesis advisor: Hywel Morgan ORCID iD
Thesis advisor: Edward James ORCID iD

University divisions

Download statistics

Downloads from ePrints over the past year. Other digital versions may also be available to download e.g. from the publisher's website.

View more statistics

Atom RSS 1.0 RSS 2.0

Contact ePrints Soton: eprints@soton.ac.uk

ePrints Soton supports OAI 2.0 with a base URL of http://eprints.soton.ac.uk/cgi/oai2

This repository has been built using EPrints software, developed at the University of Southampton, but available to everyone to use.

We use cookies to ensure that we give you the best experience on our website. If you continue without changing your settings, we will assume that you are happy to receive cookies on the University of Southampton website.

×