Intrahepatic signals involved in the recruitment and differentiation of IL-17 secreting T cells and Regulatory T cells i

Lead Research Organisation: University of Birmingham
Department Name: Immunity and Infection

Abstract

Liver disease is increasing in Western countries and there are nowhere near enough organ donors to treat the number of patients with end-stage liver disease. Chronic hepatitis leads to scarring (cirrhosis) and liver failure and the only effective treatment is liver transplantation. Lymphocytes are a type of white blood cell and an important part of the body‘s immune system. They normally fight infections but if they persist or become inappropriately activated they can cause liver injury as part of chronic hepatitis or inflammation that leads to scarring and liver failure. Lymphocytes are directed to sites of liver inflammation by molecules called chemokines that are produced in response to inflammation. These chemokines are specific in that they can direct the entry/ recruitment of particular groups of lymphocytes to specific tissues. These groups include damaging effector lymphocytes and regulatory cells which damp down inflammation leading to resolution of damage. The outcome of liver injury depends on the balance between these inflammatory and anti-inflammatory regulatory lymphocytes which in turn will be determined by a) which types of cells are recruited to the liver from blood b) signals within the liver that shape the survival, activation and expansion of specific lymphocytes. Shifting the balance towards anti-inflammatory regulatory cells may prevent liver inflammation and promote resolution thereby reversing progression to fibrosis/cirrhosis. We would like to investigate the molecular signals involved and then exploit our knowledge to develop novel therapeutic approaches using anti-inflammatory regulatory lymphocytes as cell therapy to treat chronic liver diseases.

Technical Summary

Hypothesis

Intrahepatic signals regulate the balance of effector regulatory networks in the human liver by shaping the recruitment and differentiation of functional t cells subsets.
The outcome of liver injury depends on the balance between inflammatory effector and anti-inflammatory response. In the proposed fellowship I will investigate this balance in human inflammatory liver disease by 1) defining signals that recruit and position Treg, Th1, Th22 and Th17 cells within the inflamed liver 2) defining the effect of the liver microenvironment and particularly dendritic cells and stromal cells on T cell survival and differentiation. I will also study the trafficking of autologous regulatory T cells after intravenous injection into patients with liver disease using MRI to determine homing and survival of Treg in vivo.
Aims and Design
1. Intra-hepatic signals regulate T cell subset recruitment to the human liver
I will use flow-based assays to determine endothelial and stromal cell factors that promote selective recruitment of effector versus regulatory T cells to the human liver. I will develop a human tissue assay to study post-endothelial migration of T cells in viable liver ex vivo in real-time. T cells and liver cells will be labelled with different fluorchromes and the migration of T cells through tissue tracked using Nipkov spinning-disc confocal microscopy.
2. Hepatic do stromal cells and dendritic cells regulate T cell survival and differentiation
I will study intrahepatic signals including those mediated through stromal cells that modulate T cell survival and differentiation using co-culture assays in which different cytokines and differentiation factors are added. The survival, plasticity and differentiation of Th1, Th17 and Th22 cells will be studied in Prof Paul Klenerman‘s laboratory in Oxford and the role of specific Th17 and Th122 differentiation factors will involve a collaboration with Dr Mark Veldhoen in Cambridge.
3. Homing of regulaty T cells in patients with autoimmune liver disease
The Birmingham NIHR Liver BRU has facilities and expertise to carry out cell therapy clinical trials. Within the BRU we are developing MRI to track infused cells in patients. I will use this technique to track infused autologous Treg isolated from the blood of patients with autoimmune liver disease. I have defined GMP-compliant methods for Treg isolation I will now optimise MRI labelling in collaboration with Prof Hannon (Centre for Physical Sciences of Imaging in the Biomedical Sciences) who has developed novel MRI labels for cell imaging. I will record migration and localization of infused Treg using our research-dedicated 3T MRI.
This project will provide an outstanding opportunity to continue my development as a translational researcher in immunology putting me at the forefront of cell-based therapy for liver disease.

Publications

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Bhogal RH (2018) The Reactive Oxygen Species-Mitophagy Signaling Pathway Regulates Liver Endothelial Cell Survival During Ischemia/Reperfusion Injury. in Liver transplantation : official publication of the American Association for the Study of Liver Diseases and the International Liver Transplantation Society

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Chi H (2016) Flares during long-term entecavir therapy in chronic hepatitis B. in Journal of gastroenterology and hepatology

 
Description IL-17 secreting cells in human liver
Amount £86,000 (GBP)
Organisation Novartis 
Sector Private
Country Global
Start 09/2008 
End 09/2010
 
Description Positioning and funciton of Human Th17 cell in inflamed liver (University Hospital Birmingham Charity)
Amount £70,000 (GBP)
Organisation Queen Elizabeth Hospital Birmingham Charity (QEHB) 
Sector Charity/Non Profit
Country United Kingdom
Start 09/2010 
End 02/2012
 
Title Regulatory T cells in the human liver 
Description Lab based investigation at present and plan to move to translational proof of concept of Treg tracking to human liver in future. Currently generating GMP grade Treg for future cell therapy. 
Type Of Material Model of mechanisms or symptoms - in vitro 
Provided To Others? No  
Impact N/A 
 
Description Accumulation of NKT cells in Progressive Non Alcoholic Steato Hepatitis 
Organisation Duke University Medical Centre
Country United States 
Sector Academic/University 
PI Contribution Phenotyping of Human NKT population in normal and diseaase
Collaborator Contribution Understanding of NKT in human liver disease
Impact Accumulation of NKT cells in progressive NASH,. (Hepatology. 2010 Jun;51(6):1998-2007)
Start Year 2009
 
Description CD8aa and IL-22 cells in HCV 
Organisation University of Oxford
Department Peter Medawar Building for Pathogen Research
Country United Kingdom 
Sector Academic/University 
PI Contribution Liver infiltrating lymphocytes isolation and phenotyping
Collaborator Contribution Collaborative works with liver infiltrating lymphocytes. This collaboration results in Blood and Frontiers in Immunology manuscripts.
Impact • Lucy J Walker, Yuhoi H Kang, Matthew O Smith, Hannah Tharmalingham, Ye Htun Oo, Thomas J Scriba, Willem A Hanekom, Georg M Lauer, David H Adams , Eleanor Barnes, Paul Klenerman. Human CD8aaT cells are a CD161++ subset with distinct functional characteristics. (submitted to Gastroenterology) • Yu-Hoi Kang, Eva Billerbeck, Vicki Fleming, Stefanie Grafmueller, Lucy Walker, Chris B. Willberg, Eleanor J. Barnes, Anisha Bhagwanani, Ye Htun Oo, Hubert E Blum, David H. Adams, Robert Thimme, Paul Klenerman. Recruitment of IL-22 secreting CD4+ T cells to the liver in hepatitis C. (submitted to Hepatology)
Start Year 2009
 
Description Gamma Delta cells in HCV 
Organisation University of Birmingham
Department Human Tissue Biorepository
Country United Kingdom 
Sector Academic/University 
PI Contribution To expend intracellular T cell network experiments
Collaborator Contribution Cellular culture and protein expression
Impact In progress
Start Year 2012
 
Description HCV vaccination 
Organisation University of Oxford
Department Peter Medawar Building for Pathogen Research
Country United Kingdom 
Sector Academic/University 
PI Contribution Second UK centre for HCV Therepeutic vaccination in Genotype 1 patients
Collaborator Contribution I contributed both HCV and normal healthy volunteer from Birmingham for this study. This study resulted in Hepatology and Science Translational Medicine manuscripts.
Impact Five patients from Birmingham vaccinated
Start Year 2009
 
Description Osteopontin promote fiborsis progression in NAFLD 
Organisation Duke University
Department Department of Hepatology
Country United States 
Sector Academic/University 
PI Contribution Immunohistochemical staining of Osteopontin in human liver tissue
Impact Osteopontin is induced by Hedgehog Pathway Activation and Promotes Fibrosis Progression in Nonalcoholic Steatohepatitis. Hepatology Osteopontin is induced by hedgehog pathway activation and promotes fibrosis progression in non-alcoholic steatohepatitis. Hepatology 2010 (in press DOI: 10.1002/hep.23998).
Start Year 2010
 
Description Th17 and AHR ligands 
Organisation Babraham Institute
Country United Kingdom 
Sector Private 
PI Contribution We performed staining but PI in Cambridge has left to Portugal.
Collaborator Contribution No output as PI from Cambridge left.
Impact Not started yet
Start Year 2010
 
Description Th17 and Tc17 in Human Hepatitis C infection 
Organisation University of Oxford
Department Peter Medawar Building for Pathogen Research
Country United Kingdom 
Sector Academic/University 
PI Contribution Phenotyping of freshly isolated Liver infiltrating lymphocytes
Collaborator Contribution Understanding of IL-17 secreting lymphocytes in human hepatitis C liver
Impact Analysis of CD161 expression on human CD8+ T cells defines a functional subset with tissue-homing properties; (Proc Natl Acad Sci U S A. 2010 Feb 16;107(7):3006-11)
Start Year 2008
 
Description United Kingdom Autoimmune Hepatitis Consortium (UK-AIH) 
Organisation National Institute for Health Research
Department NIHR Newcastle Biomedical Research Centre
Country United Kingdom 
Sector Academic/University 
PI Contribution National Immunology lead for UK-AIH which suits well with current liver tolerance, Regulatory T cell project
Collaborator Contribution 500 patients nation wide Newcastle Led as sponsor I am leading immunology nation wide and Birmingham Local PI.
Impact Not yet, ongoing for next 2 years
Start Year 2014
 
Description American Association of Study of Liver Disease Annual Meeting 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Geographic Reach International
Primary Audience Health professionals
Results and Impact Oral Presentation in 2008 on Regulatory T cell work Presendential Plenary Presentation in 2009 on Th17 work

Collaboration
Year(s) Of Engagement Activity 2008,2009
 
Description Association of Physician Annual Meeting 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Geographic Reach National
Primary Audience Health professionals
Results and Impact Oral presentation on Distinct role of Regulatory T cells in inflamed human liver

Clinical cell based therapy potentail
Year(s) Of Engagement Activity 2008
 
Description British Association of Study of Liver Disease Annual Meeting 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? Yes
Geographic Reach National
Primary Audience Health professionals
Results and Impact Poster presentation at BASL meeting

Collaboration and understanding the impact of therapeutic potential for Regulatory T cells
Year(s) Of Engagement Activity 2006,2007,2008
 
Description British Sceince Festival to Public in Westmidlands 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Disseminate information to public on HCV vaccination work which is now being carried out in collaboration with Oxford

Awareness of public to liver disease
Year(s) Of Engagement Activity 2010
 
Description EASL Immunology Meeting 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? Yes
Geographic Reach International
Primary Audience Health professionals
Results and Impact Phenotype and recruitment of Regulatory T cells in inflamed human liver

Collaboration, Learning new methodolgy
Year(s) Of Engagement Activity 2008
 
Description Gordon Immunology Conference 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? Yes
Geographic Reach International
Primary Audience Health professionals
Results and Impact Human Liver Infiltraing Regulator T cells

Collboration
Year(s) Of Engagement Activity 2008
 
Description MAIR conference 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Geographic Reach Regional
Primary Audience Health professionals
Results and Impact Poster presentation on human Treg work

collaboration
Year(s) Of Engagement Activity 2008
 
Description Medical Research Society Annual Meeting 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Geographic Reach National
Primary Audience Health professionals
Results and Impact Poster presentation on human liver infiltrating Treg

Collaboration
Year(s) Of Engagement Activity 2008
 
Description Medical Student Teaching on Immune Tolerance 2006,2007,2008,2009 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? Yes
Geographic Reach Local
Primary Audience Health professionals
Results and Impact Part of Medical Student's teaching on Immunology, Autoimmune hepatitis and role of Regulatory T cells

Student's understanding of Immunological tolerance and consequence of brakdown in toleranc
Year(s) Of Engagement Activity 2006,2007,2008,2009
 
Description Medical Students and BMed Science teaching 2012 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Type Of Presentation Keynote/Invited Speaker
Geographic Reach Regional
Primary Audience Postgraduate students
Results and Impact local teaching programme participation

To find BMed science student for lab project.
Year(s) Of Engagement Activity 2012
 
Description School Visit 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact Liver Transplantation ethics to grammer school students

Spread message on important of liver disease, transplantion and liver research
Year(s) Of Engagement Activity 2010
 
Description UK Adhesion Society 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Geographic Reach National
Primary Audience Health professionals
Results and Impact Oral presentation on Regularoty T cells role in human liver inflammation

Collaboration
Year(s) Of Engagement Activity 2009