Investigating triggering events in T&B Lymphocytes
Lead Research Organisation:
University of Oxford
Department Name: RDM Radcliffe Department of Medicine
Abstract
T and B lymphocytes are the 2 major cell types of the adaptive immune system, working together to coordinate a specific and long-lasting response to infectious agents and cancer. Both cell types sense their environment through surface proteins called receptors, which are responsible for detecting foreign material in the body and initiating downstream signalling to kick-start the immune response.
Despite extensive research, early events in T and B cell signalling are still not well understood. These early processes are known collectively as receptor triggering, and act as the first "go" or "no-go" checkpoint of the adaptive immune system. After receptor triggering occurs, signalling molecules interacting with the receptor coordinate downstream signalling pathways which ultimately lead to a change in behaviour of the cell.
My project will involve using super high-resolution microscopy to study these events in live cells.
I will first create model cell lines with fluorescent receptors and signalling molecules of interest by using CRISPR-Cas9 genome editing and lentiviral transduction methods. These cells will then be imaged using TIRF microscopy on different types of supported lipid bilayers, mimicking the surface of an antigen presenting cell. To complement this, I will also be analysing the stoichiometry of these receptors, looking in particular at how this changes from a resting state to activated and how proximal signalling molecules interact with the receptors.
Further work will focus on refining these imaging techniques to eventually study receptor triggering at the single-molecule level, which will address important questions about receptor sensitivity to agonist antigens. I will also investigate the role of other proximal signalling molecules and how mutations in key binding sites affect receptor triggering.
In the short term, these projects will provide insight into the exact spatial and temporal mechanisms behind receptor triggering, increasing our understanding of this fundamental first point of regulation in the adaptive immune system. In a wider context, understanding the dynamics and regulation of receptor triggering could eventually lead to the development of drugs for controlling the behaviour of T and B cells, which would be a huge benefit to many patients struggling with an autoimmune disease or supressed immune system.
Despite extensive research, early events in T and B cell signalling are still not well understood. These early processes are known collectively as receptor triggering, and act as the first "go" or "no-go" checkpoint of the adaptive immune system. After receptor triggering occurs, signalling molecules interacting with the receptor coordinate downstream signalling pathways which ultimately lead to a change in behaviour of the cell.
My project will involve using super high-resolution microscopy to study these events in live cells.
I will first create model cell lines with fluorescent receptors and signalling molecules of interest by using CRISPR-Cas9 genome editing and lentiviral transduction methods. These cells will then be imaged using TIRF microscopy on different types of supported lipid bilayers, mimicking the surface of an antigen presenting cell. To complement this, I will also be analysing the stoichiometry of these receptors, looking in particular at how this changes from a resting state to activated and how proximal signalling molecules interact with the receptors.
Further work will focus on refining these imaging techniques to eventually study receptor triggering at the single-molecule level, which will address important questions about receptor sensitivity to agonist antigens. I will also investigate the role of other proximal signalling molecules and how mutations in key binding sites affect receptor triggering.
In the short term, these projects will provide insight into the exact spatial and temporal mechanisms behind receptor triggering, increasing our understanding of this fundamental first point of regulation in the adaptive immune system. In a wider context, understanding the dynamics and regulation of receptor triggering could eventually lead to the development of drugs for controlling the behaviour of T and B cells, which would be a huge benefit to many patients struggling with an autoimmune disease or supressed immune system.
Organisations
People |
ORCID iD |
| Simon Davis (Primary Supervisor) | |
| Caitlin O'Brien-Ball (Student) |