Award details

Single molecule studies of protein organisation and triggering on live T cells

Reference	BB/D000467/1
Principal Investigator / Supervisor	Professor David Klenerman
Co-Investigators / Co-Supervisors	Professor Simon Davis, Professor William/bill FitzGerald, Dr Liming Ying
Institution	University of Cambridge
Department	Chemistry
Funding type	Research
Value (£)	429,160
Status	Completed
Type	Research Grant
Start date	01/12/2005
End date	30/04/2009
Duration	41 months

Abstract

While the proteins present on the T cell surface are now known, there is less information about how they are organised and how their organization changes, if at all, during the very earliest stages of T cell triggering. This is essentially because these early events occur on a length scale below the resolution of optical microscopy. One way to address this issue is to perform single molecule fluorescence measurements on individual molecules on the cell surface using two colour excitation and detection, so that it is possible to follow two different types of molecules simultaneously and detect interaction using coincidence fluorescence methods. We propose to exploit this method firstly to map the interactions between the key proteins on the surface of the T cell in its resting state. We will then study a model cell-contact based on the binding of CD28 to surface bound antibodies and follow the initial changes in protein reorganisation on the cell surface. This will be done using a two-colour total internal reflection microscope. This work will result in a general method to probe protein-protein interactions on cell surfaces and to follow reorganisation in structure on triggering as well as providing new molecular insights into T cell activation.

Summary

White blood cells play a key role in our immune response, responsible for clearing the body of infection. T cells are one type of white blood cell that examines the surfaces of cells to see if they have been infected. While we understand many of the events that lead to identification of an infected cell, we know less about the initial events that trigger the entire process leading to a response by the white blood cell. This is of great importance since if the testing of cells goes awry it can result in many diseases such as rheumatoid arthritis and multiple sclerosis. Since the initial events are hidden below the resolution of an optical microscope, it is necessary to track individual molecules to reveal their details by using sensitive fluorescence detection. We propose to study intially the interactions between the key proteins on the T cell surface, and then study how they reorganise during the initial processes of triggering by tracking single molecules. This will provide new insights into the molecular basis of triggering and may result in new ways to control immune responses to prevent disease.

Committee	Closed Committee - Biochemistry & Cell Biology (BCB)
Research Topics	Immunology, Technology and Methods Development
Research Priority	X – Research Priority information not available
Research Initiative	X - not in an Initiative
Funding Scheme	X – not Funded via a specific Funding Scheme

I accept the terms and conditions of use (opens in new window)

export PDF file

back to list new search