Generation of antigen specific regulatory T cells for ...Document Transcript
Biomedical Research Centre
Guy’s & St Thomas’ NHS Foundation Trust and King’s College
Project Proforma 2009
Note: Projects should be supervised by a basic scientist and clinician
Clinical Training Fellow
4 year PhD Studentship Programme X
3 year PhDs for Nurses/Midwives/Allied Healthcare Professionals
Names of project supervisors:
Dr Timothy Tree
Dr Jake Powrie
Name of the Department/Division where the research project would be
undertaken in KCL or Trust (including NHS strategic partner Trusts):
Department of Immunobiology / DIIID KCL
Diabetes and Endocrine Unit / GSTT
Name of the BRC research theme supporting the project:
Infection and Immunity Theme
Name of BRC cross cutting discipline:
Title of research project:
Generation of antigen specific regulatory T cells for therapeutic use.
Abstract of the research project:
Current efforts to prevent or reverse the severe immunopathology associated with
autoimmune disease and graft rejection rely upon the use of non-specific immune
suppressive drugs that carry the risk of serious associated side effects, such as
infection and cancer. What is required in these fields is a therapeutic approach that
offers the real possibility of a restoration of long-lasting immunological tolerance. A
series of recent seminal discoveries in immunobiology has clearly established the
existence of an immune cell type whose function is the regulation of the immune
response (Treg) and hence the maintenance of immunological tolerance. Critically,
these studies highlighted the importance of antigen specificity: antigen-specific Tregs
have far greater potency in preventing organ rejection and autoimmune diseases
when compared to those derived from polyclonal populations1. There is a clear
translational pathway from this discovery to the clinic, which could ultimately lead to
the development and use of protocols that enhance Treg function or number in
individuals at risk of developing autoimmune disease.
Recently, we have made substantial inroads into the characterization of a population
of naturally occurring regulatory T cells with relevance to the prototypical T cell
mediated autoimmune disease type 1 diabetes, which we term IL-10 secreting islet-
specific (ISIS) Tregs. In 2004 we demonstrated a link between biased production of
these Tregs and both the late onset of the T1D and the non-diabetic state, implying
that ISIS Tregs have a protective role2. In the intervening period cloning of ISIS
Tregs from non-diabetic individuals has confirmed (i) their potent suppressive
qualities and (ii) ease of expansion in vitro in substantial quantities, making them a
promising candidate for therapeutic exploitation. In addition, we have identified a
similar population of graft specific T cells present in recipients of islet allografts who
remain insulin independent following transplantation, suggesting that a similar
population of cells may play a role in transplant tolerance3.
Aims. In this project we will examine the potential to generate populations of IL-10
secreting Tregs in vitro from naïve T cell populations and investigate their stability
and therapeutic potential.
Specific Aim 1. To investigate protocols to generate populations of polyclonal Tregs
from the naïve repertoire with similar characteristics to the naturally occurring ISIS
Specific Aim 2. To implement the protocols developed in Aim 1 to generate clinical
grade ISIS Treg lines/clones from individuals with recent onset T1D.
Specific Aim 3. To investigate the influence of gene polymorphisms in T1D
susceptibility loci (including polymorphisms in the IL-10, CD25 and insulin genes) on
the generation of IL-10 secreting Tregs.
Specific Aim 4. To investigate the stability and therapeutic potential of in vitro
generated ISIS Tregs using a range of in vitro assay systems and humanised animal
Brusko, T.M., Putnam, A.L., & Bluestone, J.A., Human regulatory T cells: role in autoimmune disease
and therapeutic opportunities. Immunol Rev 223, 371-390 (2008).
Arif, S. et al., Autoreactive T cell responses show proinflammatory polarization in diabetes but a
regulatory phenotype in health. J Clin Invest 113 (3), 451-463 (2004).
Huurman, V.A. et al., Allograft-specific cytokine profiles associate with clinical outcome after islet cell
transplantation. Am J Transplant 9 (2), 382-388 (2009).
Outline the Departmental/Divisional research training and inhouse research
support in relation to this project:
Training in research
The Programme in Infection and Immunity (PII) laboratories are extremely well-
equipped, having been opened as recently as 2004. Core facilities include flow
cytometry; Q-RT-PCR; confocal microscopy; IT-room; cell-culture; and Cat-3 areas.
Core-staff supervising these facilities are charged with training new staff in their
appropriate use of relevant techniques. Each researcher has a dedicated desk area
with full e-access. In addition, a successful applicant would join a wider group of
researchers based at KCL investigating Enhanced Cell Therapy for Cancer,
Autoimmunity and Transplantation funded in part by funded by an Interdisciplinary
Programme in Translational Research award by the NIMR, of which the applicant (Dr
Timothy Tree) is a Principal Investigator. This Programme will provide the researcher
with all specialist equipment required to adapt protocols developed in the research
environment to those suitable for the generation clinical grade T cells suitable for use
in patients (e.g. GMP grade cell isolation, manipulation and culture).
Training in scholarship
The PII research staff attend a weekly seminar and have lunch with the outside
speaker. There is also mandatory attendance at the weekly PII Research Update
Conference (RUC), where graduates, postgraduates and fellows alike present their
data once or twice per year, so as to receive wide-ranging feedback and hone
presentation skills. Research staff also attend journal clubs (both departmental and
divisional); individual lab meetings; and research retreats, as well as periodic
“specialist meetings” such as those in immunoregulation and transplant tolerance.
They are also encouraged to attend the bi-weekly Biomedical Research Forum.
Researchers are strongly encouraged (via payment of fees) to attend the British
Society of Immunology Congress, and it is common for them to be sent on specialist-
skills courses, such as those run through the Wellcome Trust Sanger Centre. The
Division also organises additional programmes for researchers, such as the Nomura-
enterprise seminar series, and one-day workshops in aspects of immunology and
virology. Formal lecture programmes in cellular and molecular immunology, as well
as all aspects of clinical immunology are conducted as components of our two MSc
Programmes (Immunology and Medical Immunology) and attendance of these on an
ad hoc basis is encouraged.
Relevance of the project to the research theme and cross cutting discipline:
The Infection and Immunity theme addresses the autoimmune processes that lead to
Type 1 diabetes as part of its programme of research, and includes a considerable
thrust on strategies to halt or prevent islet destruction. These efforts include projects
funded by an Interdisciplinary Programme in Translational Research award by the
NIMR to establish a Programme in Enhanced Cell Therapy for Cancer, Autoimmunity
and Transplantation. This Programme is a multi-disciplinary, multi-theme BRC body
incorporating clinicians and scientists in the strategic partners (KCH and GST) and
cross-cutting disciplines (including Imaging Sciences) and aims to take cellular
therapies for a range of clinical settings including autoimmunity and organ
transplantation to first-in-man studies and beyond.
In addition, this project proposes to investigate a link between polymorphisms at T1D
susceptibility loci such as IL-10, CD25 and Ins and the ability to generate ISIS Tregs.
This forms part of a wider body of research investigating T1D genotype-immune
phenotype relationships funded by a Centre Grant by the Juvenile Diabetes
Foundation (Diabetes- Genes, Autoimmunity and Prevention Centre) and includes
researchers both in the Department of Immunobiology at KCL (Professor Mark
Peakman and Dr Tim Tree) and at the University of Cambridge (Professor John Todd
and Professor Linda Wicker).
Please provide project supervisors contact details - which can be used by
potential applicants wishing to discuss the project:
Dr Timothy Tree
Department of Immunobiology
Programme of Infection and Immunity
King's College London School of Medicine
at Guy's, King's College and St Thomas' Hospitals
New Guy's House
London SE1 9RT
Tel: 020 71881182
Fax: 020 7188 3385
Dr J K Powrie MD FRCP
Consultant, Honorary Senior Lecturer and Clinical Lead in Diabetes and
Diabetes and Endocrine Unit
Great Maze Pond
London SE1 9RT
Tel: 020 71881912
Fax: 020 71881926