The molecular and cellular basis of complement-mediated T helper 1 (TH1) differentiation and regulation

Lead Research Organisation: King's College London
Department Name: Immunology Infection and Inflam Diseases

Abstract

A group of white cells called T lymphocytes forms a major part of our immune system and self-defence against dangerous infections. T lymphocytes seek and destroy pathogen-infected tissue and cells. However, these lymphocytes sometimes wrongly recognise proteins on our own tissues: This can cause destructive conditions such as rheumatoid arthritis and multiple sclerosis. Because T lymphocytes have such potent ability to be either beneficial or dangerous, their function is tightly regulated. To fight infections, these cells produce a cytokine called IFN-gamma. One of the control mechanisms to regulate the production of IFN-gamma is the induction of another cytokine, IL-10, within the same T cell, which then ?shuts down? IFN-gamm; expression. However, how exactly the T cell regulates the appropriate production of IFN-gamma vs. IL-10 is not understood. We have recently discovered that a protein called CD46 (which is present on the surface of T lymphocytes) is an important trigger for the switching of IFN-gamma to IL-10 in human T cells by sending appropriate signals into the cell. In addition, we have made the exciting observation that our finding is important for understanding a major human disease, rheumatoid arthritis (RA): We observed that the CD46 signals transduced do not work properly in T cells from RA patients (they do not shut down IFN-gamma). We now aim at finding out what exactly happens on a molecular level inside the cells when CD46 is activated on the cell surface of a healthy T cell. Understanding these ?signaling events? within healthy T cells will then allow us to compare those to data to signals from RA patients? T cells and define exactly were things ?are going wrong?. The understanding of this induction pathway will then lay the groundwork to understand the pathogenesis of RA and thereby to develop better diagnostics and therapies. The main investigator (CK) of this study is a leading authority in the functions of CD46 and regulatory lymphocytes. She has recently relocated from the US to the MRC Centre for Transplantation at King?s College and will now collaborate with outstanding experts (AC) in the RA field and in cytokine regulation (PL) on this project. Because King?s emphasizes strongly the crosstalk between basic research and clinical/patient work, CK will be in a position to rapidly interpret the findings for patient benefit. The proposed funding will help establish this promising body of work in the UK.

Technical Summary

Control of IFN-gamma-secreting CD4+ TH1 effector T cells is vital for the prevention of immunopathologies during and after infection and unwanted responses against self or innocuous antigens. IL-10 is a key cytokine mediating negative control of TH1 responses. Although IL-10 is produced by several cell types and, thus, many cells can ?extrinsically? suppress TH1 responses, a novel concept now suggests that the major TH1 suppression pathway may be represented by ?intrinsic? regulation: A negative feedback loop working via the timely coinduction of IL-10 in addition to IFN-gamma within TH1 cells. Although the importance of IFN-gamma to IL-10 switching is acknowledged, the molecular pathways regulating these two cytokines in TH1 cells are undefined.
We identified the complement regulator CD46 as key receptor in regulating IFN-gamma and IL-10 production in TH1 cells (Cardone et. al, Nature Immunology, 2010). We demonstrated that CD46 not only drives IFN-gamma production and effector function of human TH1 cells but also induces the switch to IL-10 secretion and into the regulatory phase in an IL-2-dependent fashion: In the presence of low IL-2 CD46 induces almost exclusively IFN-gamma+ effector TH1 cells while high environmental IL-2 then switches these cells from an IFN-gamma+/IL-10+ intermediate finally into IL-10+ T cells, with both of the latter populations being suppressive. Importantly, we showed that this CD46-mediated switch is defective in rheumatoid arthritis (RA) patients: T cells from these patients lack the IL-10+ cell subpopulation, produce up to 20 times more IFN-gamma compared to those from healthy individuals and never enter the regulatory phase. Thus, we hypothesize that CD46-transduced signals play a vital role in the induction and resolution of human TH1 responses and that disturbance in CD46 signals is associated with RA. This application aims now at delineating the molecular signals regulating IFN-gamma/IL-10 switching in human TH1 cells. We propose that each of the CD46/IL-2-mediated sequential phases in the TH1 cell ?life cycle? (IFN-gamma+, the IFN-gamma+/IL-10+ and IL-10+) are defined by the expression of specific genes and miRNAs as well as distinct changes in the chromatin environment. We will identify those changes and generate a comprehensive transcriptomic signature of each of the TH1 induction and regulation phases present in T cells from healthy donors (in comparison with T cells from RA patients). This ?molecular landscape? will then provide an experimental framework to identify single proteins and pathways required for IFN-gamma/IL-10 switching, and to uncover the defect(s) in TH1 regulation in RA.
 
Description Renal Project Board Steering Committee
Geographic Reach National 
Policy Influence Type Membership of a guidance committee
 
Description BTCURE: Understanding aberrant adaptive immunity mechanisms in human chronic immune-mediated diseases: rheumatoid arthritis, systemic lupus erythematosus and inflammatory bowel disease.
Amount £680,000 (GBP)
Organisation European Commission 
Department Seventh Framework Programme (FP7)
Sector Public
Country European Union (EU)
Start 04/2011 
End 03/2017
 
Description Wellcome Trust New Investigator
Amount £1,540,000 (GBP)
Organisation Wellcome Trust 
Sector Charity/Non Profit
Country United Kingdom
Start 04/2014 
End 03/2019
 
Title Protocol for the proteolytic cleavage of C3 by cathepsin L 
Description This output describes in detail the cleavage and activation of human C3 by cathepsin L - a method discovered in my laboratory. 
Type Of Material Technology assay or reagent 
Year Produced 2013 
Provided To Others? Yes  
Impact Not yet measurable 
URL http://www.bio-ptotocol.org/e1205
 
Title Gene arrays of T cells from patients with CD46 deficiency 
Description We have generated gene array analyses from rare patients with deficiency in CD46 expression (non-activated and 2 hr time point post TCR, TCR and CD28 and TCR and CD46 activation). 
Type Of Material Database/Collection of data 
Provided To Others? No  
Impact Data are in the progress of being published on OA data bases 
 
Description Assessment of CD46-mediated IFN-gamma to IL-10 switching as diagnostic tool for TNF weaning in RA 
Organisation King's College London
Department Department of Academic Rheumatology
Country United Kingdom 
Sector Academic/University 
PI Contribution Development of an in vivo assay to assess if RA patents can be tapered of anti-TNF therapy
Collaborator Contribution My partners contribute with the provision of clinical patients samples, data analyses etc.
Impact Ongoing. First data expected end 2013
Start Year 2010
 
Description Characterization of a novel convertase-independent intracellular complement activation pathway 
Organisation Imperial College London
Department Centre for Complement and Inflammation Research
Country United Kingdom 
Sector Academic/University 
PI Contribution We have discovered a novel intracellular complement activation pathway and are spear-heading the further characterization of this pathway in collaboration with the listed investigators.
Collaborator Contribution Our collaborators are contributing with expertise, reagents, crystal structure generation and pertinent animal models to this study.
Impact Manuscript published in 'Immunity'. Currently working on a follow-up manuscript.
Start Year 2011
 
Description Characterization of a novel convertase-independent intracellular complement activation pathway 
Organisation King's College London
Department Randall Division of Cell & Molecular Biophysics
Country United Kingdom 
Sector Academic/University 
PI Contribution We have discovered a novel intracellular complement activation pathway and are spear-heading the further characterization of this pathway in collaboration with the listed investigators.
Collaborator Contribution Our collaborators are contributing with expertise, reagents, crystal structure generation and pertinent animal models to this study.
Impact Manuscript published in 'Immunity'. Currently working on a follow-up manuscript.
Start Year 2011
 
Description Characterization of a novel convertase-independent intracellular complement activation pathway 
Organisation Spanish National Research Council (CSIC)
Department Biological Research Center
Country Spain 
Sector Private 
PI Contribution We have discovered a novel intracellular complement activation pathway and are spear-heading the further characterization of this pathway in collaboration with the listed investigators.
Collaborator Contribution Our collaborators are contributing with expertise, reagents, crystal structure generation and pertinent animal models to this study.
Impact Manuscript published in 'Immunity'. Currently working on a follow-up manuscript.
Start Year 2011
 
Description Characterization of a novel convertase-independent intracellular complement activation pathway 
Organisation University Medical Center Freiburg
Country Germany 
Sector Hospitals 
PI Contribution We have discovered a novel intracellular complement activation pathway and are spear-heading the further characterization of this pathway in collaboration with the listed investigators.
Collaborator Contribution Our collaborators are contributing with expertise, reagents, crystal structure generation and pertinent animal models to this study.
Impact Manuscript published in 'Immunity'. Currently working on a follow-up manuscript.
Start Year 2011
 
Description Characterization of a novel convertase-independent intracellular complement activation pathway 
Organisation University Medical Center Utrecht (UMC)
Country Netherlands 
Sector Academic/University 
PI Contribution We have discovered a novel intracellular complement activation pathway and are spear-heading the further characterization of this pathway in collaboration with the listed investigators.
Collaborator Contribution Our collaborators are contributing with expertise, reagents, crystal structure generation and pertinent animal models to this study.
Impact Manuscript published in 'Immunity'. Currently working on a follow-up manuscript.
Start Year 2011
 
Description Characterization of a novel convertase-independent intracellular complement activation pathway 
Organisation Washington University in St Louis
Department Division of Rheumatology
Country United States 
Sector Academic/University 
PI Contribution We have discovered a novel intracellular complement activation pathway and are spear-heading the further characterization of this pathway in collaboration with the listed investigators.
Collaborator Contribution Our collaborators are contributing with expertise, reagents, crystal structure generation and pertinent animal models to this study.
Impact Manuscript published in 'Immunity'. Currently working on a follow-up manuscript.
Start Year 2011
 
Description Characterization of the molecular crosstalk between CD46 and IL-2R during Th1 induction and regulation 
Organisation King's College London
Department Division of Immunology, Infection & Inflammatory Diseases (DIIID)
Country United Kingdom 
Sector Academic/University 
PI Contribution We have developed successfully an approach to differentiate between the CD46 and the IL-2R-mediated signals in Th1 induction and are using this approach to define the single contributions as well as the crosstalk of these receptor-mediated pathways in IFN-gamma induction and IL-10 switching.
Collaborator Contribution Our collaborators contribute with reagents, expertise and pertinent experiments. Results and subsequent experimental strategies are discussed during regular meetings.
Impact U.S. Provisional Patent Application U.S.S.N.61/868,016 with full patent application pending
Start Year 2011
 
Description Characterization of the molecular crosstalk between CD46 and IL-2R during Th1 induction and regulation 
Organisation University of Oxford
Department Sir William Dunn School of Pathology
Country United Kingdom 
Sector Academic/University 
PI Contribution We have developed successfully an approach to differentiate between the CD46 and the IL-2R-mediated signals in Th1 induction and are using this approach to define the single contributions as well as the crosstalk of these receptor-mediated pathways in IFN-gamma induction and IL-10 switching.
Collaborator Contribution Our collaborators contribute with reagents, expertise and pertinent experiments. Results and subsequent experimental strategies are discussed during regular meetings.
Impact U.S. Provisional Patent Application U.S.S.N.61/868,016 with full patent application pending
Start Year 2011
 
Description Dissecting the roles of anaphylatoxins in human Th1 induction and regulation 
Organisation Oslo University Hospital
Department Department of Immunology and Transfusion Medicine
Country Norway 
Sector Hospitals 
PI Contribution We have developed a hypothesis and preliminary data on novel roles for several complement activation fragments in human Th1/Th2 regulation and will provide further cell-based assay data.
Collaborator Contribution Our collaborators will provide unique reagents (proteins, inhibitors etc.) as well as pertinent patient blood samples (all ethics in place) and validation of new findings for in vivo importance then in mouse models.
Impact Manuscript in preparation
Start Year 2012
 
Description Dissecting the roles of anaphylatoxins in human Th1 induction and regulation 
Organisation University of Lubeck
Department Institute for Inflammation Research
Country Germany 
Sector Academic/University 
PI Contribution We have developed a hypothesis and preliminary data on novel roles for several complement activation fragments in human Th1/Th2 regulation and will provide further cell-based assay data.
Collaborator Contribution Our collaborators will provide unique reagents (proteins, inhibitors etc.) as well as pertinent patient blood samples (all ethics in place) and validation of new findings for in vivo importance then in mouse models.
Impact Manuscript in preparation
Start Year 2012
 
Description Dissecting the roles of anaphylatoxins in human Th1 induction and regulation 
Organisation University of Pennsylvania
Department Department of Pathology and Laboratory Medicine
Country United States 
Sector Academic/University 
PI Contribution We have developed a hypothesis and preliminary data on novel roles for several complement activation fragments in human Th1/Th2 regulation and will provide further cell-based assay data.
Collaborator Contribution Our collaborators will provide unique reagents (proteins, inhibitors etc.) as well as pertinent patient blood samples (all ethics in place) and validation of new findings for in vivo importance then in mouse models.
Impact Manuscript in preparation
Start Year 2012
 
Description Structural analyses on the role of complement and Notch system interactions 
Organisation University of Oxford
Department Sir William Dunn School of Pathology
Country United Kingdom 
Sector Academic/University 
PI Contribution We are contributing the functional assays and data on the in vivo importance of the complement/Notch system crosstalk
Collaborator Contribution Our collaborators provide the protein and biochemical 'side' of this project.
Impact Publication in a high impact journal (Nature Immunology 2012) and an ongoing collaboration with additional novel data likely also publishable in a high impact journal.
Start Year 2010
 
Description The crosstalk beteen complement and the IL-2 receptor system 
Organisation King's College London
Country United Kingdom 
Sector Academic/University 
PI Contribution We have observed a new function for certain complement components in the regulation of the IL-2 receptor family system and performing the cell biology experiments to dissect those
Collaborator Contribution This partner contributes unique expertise and reagents (mouse strains) to the study
Impact None yet. Manuscript submission is expected at the end of 2015.
Start Year 2014
 
Description The crosstalk beteen complement and the IL-2 receptor system 
Organisation National Institutes of Health (NIH)
Country United States 
Sector Public 
PI Contribution We have observed a new function for certain complement components in the regulation of the IL-2 receptor family system and performing the cell biology experiments to dissect those
Collaborator Contribution This partner contributes unique expertise and reagents (mouse strains) to the study
Impact None yet. Manuscript submission is expected at the end of 2015.
Start Year 2014
 
Description The role of C5 in the negative regulation of human Th1 immunity 
Organisation University of Oslo
Department Centre for Immune Regulation
Country Norway 
Sector Academic/University 
PI Contribution Collaborative experiments on new functions of C5aR and C5L2 in human T cell biology, exchange of ideas and data
Collaborator Contribution Delivery of rare C5-deficient patient samples, collaborative experiments, exchange of reagents
Impact A manuscript is in preparation. The data from this collaboration were presented at the 14th European meetimg on Complement in Human Disease and chosen for an oral presentation and outstanding oral presentation award.
Start Year 2012
 
Description The role of CD46 in key T cell metabolic pathways 
Organisation University of Basel
Country Switzerland 
Sector Academic/University 
PI Contribution We generated preliminary data on which the study is based and performed the core T cell activation experiments
Collaborator Contribution This group has performed all experiments in conjunction with T cell metabolic measurements and had critical and equal intellectual input into this project.
Impact Manuscript is under review in a high impact journal.
Start Year 2013
 
Description The role of immmune cell-derived complement in Scleroderma 
Organisation University College London
Department Division of Medicine
Country United Kingdom 
Sector Academic/University 
PI Contribution Collaborative experiments, training of a postdoctoral fellow from Dr. Abraham's laboratory (Dr. Sandra Guerra) in T cell-related techniques with emphasis on the roles of T cell-expressed complement receptors in Scleroderma.
Collaborator Contribution Delivery of Scleroderma patients samples, collaborative experiments, exchange of data, reagents and ideas.
Impact We expect outputs and outcomes by the middle/end of 2014
Start Year 2013
 
Description The role of of novel in human T cell Biology 
Organisation Nagoya University
Department Graduate School of Medicine
Country Japan 
Sector Academic/University 
PI Contribution Collaborative experiments on the role of a novel complement receptor in human T cell responses. We are assessing the novel function of this molecule mostly on human T cells.
Collaborator Contribution Dr. Takahashi's group provided the receptor KO mouse strain to us and is performing at the same time experiments to asses the role of this molecule in vivo in infection models.
Impact Data generation is progressing well, we expect a first collaborative paper in 2014.
Start Year 2013
 
Description Therapeutic targeting of complement receptors 
Organisation Apellis Pharmaceutical
Country United States 
Sector Private 
PI Contribution Experiments on the role of specific complement receptors in T cell responses in in vivo disease models.
Collaborator Contribution Generation and provision of human complement receptor targeted reagents (agonists and antagonists).
Impact U.S. Provisional Patent Application U.S.S.N.61/868,016 with full patent application pending
Start Year 2012
 
Title COMPOSITIONS AND METHODS RELATING TO C5L2 (Inventor: Claudia Kemper, co-applicants: KCL and Apellis) 
Description We discovered that C5L2 on the surface of all T cells, monocytes and stem cells can impact in cell function and differentiation. This provisional patent protects the development of a wide range of specific C5L2 agonistic and antagonistic reagents (chemical, petides, antibodies etc. for a wide range of applications (human dieseases, transplantation, stem cell development etc.) 
IP Reference US Provisional patent application 
Protection Patent application published
Year Protection Granted 2013
Licensed Commercial In Confidence
Impact Expected in 2014