Leucocyte and Pathogen Trafficking through Lymphatics in Inflammation and Immunity
Lead Research Organisation:
University of Oxford
Department Name: UNLISTED
Abstract
We are investigating how cells enter and move within lymphatic vessels so that in the future we can develop new therapies to block inflammation, prevent tumour spread, and make the delivery of vaccines to emerging infections more efficient. The lymphatic vessels form a dense network that intertwines the blood circulation and whose main functions are to maintain fluid balance and to transport cells carrying foreign antigen to the lymph nodes where they meet T and B cells and stimulate the immune response. Importantly, the lymphatics also contribute to diseases such as cancer by allowing metastatic tumours (e.g. skin and breast cancer) to spread to other parts of the body, and they act as a reservoir for malignant cells. Even though the involvement of lymphatics in disease have been known for a long time, their biology is still not well understood and so research in my group is dedicated to understanding how immune cells and tumours enter and navigate within the lymphatics. Thus far we have discovered that in inflammation, entry of immune cells carrying proteins from infectious agents depends critically on recognition of inflammation-induced protein receptors on the surface of lymphatic vessels, and that this is obligatory for the generation of the immune response to infection. We have also identified a separate mechanism that controls tumour cell entry to the lymphatics during lymph node metastasis in murine tumours and humans. We are also particularly interested in researching a molecule in lymphatics called LYVE-1 and we anticipate that further research in this area will reveal new ways to block inflammation, and to promote protective immunity to emerging infectious agents by enhancing vaccine delivery.
Technical Summary
Goals To investigate mechanisms of lymphatic trafficking in inflammation, immunity and cancer. In addition to maintaining fluid balance and tissue homeostasis the lymphatics also provide a conduit for trafficking of antigen presenting cells to lymph nodes for activation of nave T cells, and in so doing they allow a route for tumour metastasis. Recent work has focused on understanding the key molecular mechanisms regulating entry of leukocytes and tumour cells to lymphatics and their subsequent trafficking to lymph nodes. We have discovered a separate new integrin and chemokine dependent pathway for translymphatic migration of dendritic cells (DC) that functions in inflammation (1) and demonstrated its importance for dermal antigen delivery during T cell responses - identifying a new potential target for intervention (2). We have focused on two lectin-like receptors of the Link superfamily, LYVE-1 and CD44. LYVE-1 is the major HA receptor of lymphatic endothelial cells and was originally cloned and characterized in my laboratory (3). We have shown that in addition to mediation of lymphatic trafficking of DC, LYVE-1 also plays a role in lymphatic invasion in tumour metastasis and in the resolution of inflammation in a model of acute lung injury. We have identified tumour lymphatic endotherlial cell markers (TLEM) and have shown these to be important in nodal metastasis in a range of murine and human cancers (4). The crystal structure and activation mechanism of CD44, an inflammatory leukocyte HA receptor closely related to LYVE-1 and which plays a pivotal role in transmigration of activated leukocytes across inflamed vascular endothelium have been determined (5). Future research plans Future work will concentrate on the definition of the lymphatic entry points and molecular mechanisms for transmigration of different leukocyte populations. The use of microscopic imaging and structure/function analyses will allow us to determine how receptors such as LYVE-1 mediate cell trafficking in animal models of inflammation. Moreover, we will pursue the 3D structure of the LYVE-1 ectodomain and elucidate the mechanisms regulating it's interaction with hyaluronan, search for additional LYVE-1 ligands mediating leukocyte trafficking, define downstream signalling pathways mediating transendothelial migration and investigate function blocking antibodies of potential therapeutic value for inflammation and cancer. References: (1) Johnson et al. 2006 J Exp Med 203: 2763; (2) Teoh et al. 2009 J Immunol 182: 2425; (3) Gale et al 2007 Mol Cell Biol 27: 595; (4) Clasper et al. 2008 Cancer Res 68:7293; (5) Banerji et al. 2007 Nature Struct Mol Biol 14:234
Organisations
- University of Oxford (Lead Research Organisation)
- UNIVERSITY OF OXFORD (Collaboration)
- ETH Zurich (Collaboration)
- Medical University of Vienna (Collaboration)
- Regeneron Pharmaceuticals, Inc. (Collaboration)
- Wellcome Trust (Collaboration)
- University of Houston (Collaboration)
- University of Seville (Collaboration)
- University Clinic Cologne (Collaboration)
- Centre for Cooperative Research in Biomaterials (CIC BiomaGUNE) (Collaboration)
- IMPERIAL COLLEGE LONDON (Collaboration)
People |
ORCID iD |
David Jackson (Principal Investigator) |
Publications
Vieira J
(2018)
The cardiac lymphatic system stimulates resolution of inflammation following myocardial infarction
in Journal of Clinical Investigation
Sun M
(2019)
Hyaluronan Derived From the Limbus is a Key Regulator of Corneal Lymphangiogenesis.
in Investigative ophthalmology & visual science
Stanly T
(2020)
The cortical actin network regulates avidity-dependent binding of hyaluronan by the lymphatic vessel endothelial receptor LYVE-1
in Journal of Biological Chemistry
Siggins MK
(2020)
Extracellular bacterial lymphatic metastasis drives Streptococcus pyogenes systemic infection.
in Nature communications
Ravaud C
(2021)
Lymphatic Clearance of Immune Cells in Cardiovascular Disease.
in Cells
Podgrabinska S.
(2003)
Erratum: Molecular characterization of lymphatic endothelial cells (Proceedings of the National Academy of Sciences of the United States of America (December 10, 2002) 99: 25 (16069-16074))
in Proceedings of the National Academy of Sciences of the United States of America
Lu W
(2019)
TNF-derived peptides inhibit tumour growth and metastasis through cytolytic effects on tumour lymphatics.
in Clinical and experimental immunology
Johnson LA
(2021)
Hyaluronan and Its Receptors: Key Mediators of Immune Cell Entry and Trafficking in the Lymphatic System.
in Cells
Related Projects
Project Reference | Relationship | Related To | Start | End | Award Value |
---|---|---|---|---|---|
MC_UU_00008/1 | 31/03/2017 | 30/03/2023 | £2,738,000 | ||
MC_UU_00008/2 | Transfer | MC_UU_00008/1 | 31/03/2017 | 30/03/2023 | £1,821,000 |
MC_UU_00008/3 | Transfer | MC_UU_00008/2 | 31/03/2017 | 30/03/2023 | £2,257,000 |
MC_UU_00008/4 | Transfer | MC_UU_00008/3 | 31/03/2017 | 30/03/2023 | £1,459,000 |
MC_UU_00008/5 | Transfer | MC_UU_00008/4 | 31/03/2017 | 30/03/2023 | £1,346,000 |
MC_UU_00008/6 | Transfer | MC_UU_00008/5 | 31/03/2017 | 30/03/2023 | £1,660,000 |
MC_UU_00008/7 | Transfer | MC_UU_00008/6 | 31/03/2017 | 30/03/2023 | £401,000 |
MC_UU_00008/8 | Transfer | MC_UU_00008/7 | 31/03/2017 | 31/03/2024 | £2,876,000 |
MC_UU_00008/9 | Transfer | MC_UU_00008/8 | 31/03/2017 | 30/03/2023 | £2,568,000 |
MC_UU_00008/10 | Transfer | MC_UU_00008/9 | 31/03/2017 | 30/03/2023 | £2,060,000 |
MC_UU_00008/11 | Transfer | MC_UU_00008/10 | 31/03/2017 | 30/03/2023 | £1,477,000 |
Description | CRUK Grant reviewing |
Geographic Reach | National |
Policy Influence Type | Participation in a guidance/advisory committee |
Description | Research Grant Review committee Finnish Academy of Sciences 31 Jan 1 Feb 2022 |
Geographic Reach | Europe |
Policy Influence Type | Participation in a guidance/advisory committee |
Description | MRC Research Grant Streptococcal dissemination |
Amount | £592,795 (GBP) |
Funding ID | MR/L008610/1 |
Organisation | Medical Research Council (MRC) |
Sector | Public |
Country | United Kingdom |
Start | 03/2014 |
End | 03/2017 |
Description | Revealing complexity of hyaluronan-protein interactions: novel tools and insights |
Amount | £937,158 (GBP) |
Funding ID | BB/X007278/1 |
Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
Sector | Public |
Country | United Kingdom |
Start | 03/2023 |
End | 03/2026 |
Title | Characterisation of leucocyte hyaluronan surface glycocalyx and its role in lymphatic docking and entry |
Description | Imaging of hyaluronan on surface of dendritic cells in mouse tissues and in mouse and human tissue-derived dendritic cells using biotinylated VG1. Analaysis of dendritic cell trafficking in mouse inflamed skin lymphatics before/after depletion of surface hyaluronan using 4-MU and hyaluronidase. |
Type Of Material | Model of mechanisms or symptoms - mammalian in vivo |
Year Produced | 2017 |
Provided To Others? | No |
Impact | Publication Nature Immunology paper (Johnson LA et al In Press 2017). Related aspects (HA glycocalyx in mouse macrophages) published in PMID 26823460 |
Description | Blockade of LYVE-1 for prevention of skin transplant rejection |
Organisation | University of Oxford |
Department | Nuffield Department of Surgical Sciences Oxford |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Provision of mAbs, transgenic mice, advice and expertise |
Collaborator Contribution | Conduct of mouse allogeneic skin transplant experiments, advice and expertise |
Impact | The collaboration has just recently begun. Outputs are expected in the coming months |
Start Year | 2022 |
Description | Corneal lymphangiogenesis Vivien Coulson-Thomas |
Organisation | University of Houston |
Department | College of Optometry |
Country | United States |
Sector | Academic/University |
PI Contribution | Advice expertise and experimental design |
Collaborator Contribution | Performed analyses of corneal lymphangiogenesis in relationship to local hyaluronan deposition in mouse models of corneal inflammation |
Impact | Co-author Publication "Hyaluronan derived from the limbus is a key Regulator of Corneal Lymph-angiogenesis" Sun M, Puri S, Mutoji N, Coulson-Thomas YM, Hascall V, Jackson DG, Gesteira TF & Coulson- Thomas, VJ (2019) Invest. Ophthalmol. Vis. Sci. 60, 1050-1062. |
Start Year | 2018 |
Description | Corneal transplant rejection - Claus Cursiefen, Felix Bock |
Organisation | University Clinic Cologne |
Department | Department of Ophthalmology |
Country | Germany |
Sector | Academic/University |
PI Contribution | Provision of blocking mAbs and transgenic mice. Experimental design. |
Collaborator Contribution | Testing of mAbs and transgenic mice in corneal allograft rejection model. Does disruption of LYVE-1 : hyaluronan interactions in limbal lymphatics ameliorate transplant rejection and/or lymphangiogenesis ? |
Impact | In progress. Multidisciplinary - clinical (surgical), and microscopic imaging |
Start Year | 2018 |
Description | Jesus Angulo NMR collaboration |
Organisation | University of Seville |
Department | Department of Bioorganic Chemistry |
Country | Spain |
Sector | Academic/University |
PI Contribution | Provision of recombinant soluble CD44 protein for STD NMR experiments |
Collaborator Contribution | Performed STD NMR analyses |
Impact | In progress |
Start Year | 2012 |
Description | Lung lymphatics mAbs |
Organisation | Medical University of Vienna |
Department | Clinical Institute of Pathology |
Country | Austria |
Sector | Academic/University |
PI Contribution | Generation of mAbs for imaging vasculature in lung sections |
Collaborator Contribution | EM and confocal imaging of lung vasculature |
Impact | Manuscript in preparation |
Start Year | 2012 |
Description | Paul Riley collaboration - role of lymphatics in response to myocardial infarction |
Organisation | University of Oxford |
Department | Department of Physiology, Anatomy and Genetics |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Reagents, transgenic mice, advice and expertise |
Collaborator Contribution | Reagents, conduct of experiments in mouse model of MI, advice and expertise |
Impact | 3 Publications to date "The cardiac lymphatic system stimulates resolution of inflammation following myocardial infarction" Vieira JM, Norman, S, Cahill, TJ, Villa del Campo, C, Rohling, M, Barnette, DN, Johnson, LA, Greaves, DR, Carr, CA, Jackson, DG & Riley P (2018) J. Clin. Invest. 28, :3402-3412. "Tissue-resident macrophages regulate lymphatic vessel growth and patterning in the developing heart" Cahill TJ, Sun X, Ravaud C, Villa del Campo C, Klaourakis K, Lupu IE, Lord AM, Browne C, Jacobsen SEW, Greaves DR, Jackson DG, Cowley SA, James W, Choudhury RP, Vieira JM and Riley RP. (2021) Development 148 dev194563 https://doi.org/10.1242/dev.194563 "Lymphatic clearance of immune cells in cardiovascular disease". Ravaud, C, Ved, N. Jackson, DG, Vieira, JM and Riley, PR. (2021) Cells 2021, 10, 2061. https://doi.org/10.3390/cells10082061 (Special Issue "Mechanisms of Lymphatic Trafficking in Inflammation, Immunity and Metastasis"). |
Start Year | 2015 |
Description | Ralf Richter collaboration |
Organisation | Centre for Cooperative Research in Biomaterials (CIC BiomaGUNE) |
Department | Biosurfaces Research Unit |
Country | Spain |
Sector | Academic/University |
PI Contribution | Generation of protein reagents for ligand binding analysis by monitoring changes in mass and viscoelastic properties using accoustic resonance measurements (Quartz Crystal Microbalance with dissipation monitoring - QCM-D) technique and Atomic Force Spectroscopy. Intellectual input experimental design and analysis, co-authoring |
Collaborator Contribution | Expertise in physical biochemical techniques |
Impact | Publications "Analysis of CD44-hyaluronan interactions in an artificial membrane system, insights into the distinct binding properties of high and low molecular weight hyaluronan" (2010) Wolny PM, Banerji S, Gounou C, Brisson AR, Day AJ, Jackson DG, & Richter RP. J. Biol. Chem. 285, 30170-30180. "Micromechanical Analysis of the Hyaluronan-Rich Matrix Surrounding the Oocyte Reveals a Uniquely Soft and Elastic Composition" Chen, X, Bonfiglio, R, Banerji, S, Jackson, DG, Salustri, A & Richter, R (2016) Biophys. J. 110, 2779-2789. "A Single molecule assay to probe monovalent and multivalent bonds between hyaluronan and its key leukocyte receptor CD44 under force" Bano, F, Banerji, S, Howarth, M, Jackson, DG & Richter RP (2016) Sci. Rep. 6, 34176; doi: 10.103 Multidisciplinary Biochemistry, Biophysics, Immunology |
Start Year | 2016 |
Description | Regeneron collaboration |
Organisation | Regeneron Pharmaceuticals, Inc. |
Country | United States |
Sector | Private |
PI Contribution | Characterisation of phenotype of LYVE1 KO mice, generation of potential human function blocking mAbs and authoring of publication |
Collaborator Contribution | Generated transgenic mice, providing intellectual contributions and technical expertise for mAb production and testing |
Impact | Generated publication "Normal lymphatic development and function in mice deficient for the lymphatic hyaluronan receptor LYVE-1" Gale, NW, Prevo, R, Espinosa-Fematt, J, Dominguez, M, Ferguson, DJ, Yancopoulos, G. Thurston, G & Jackson, DG (2007) Mol. Cell. Biol. 27, 595-604. "Dendritic cells enter lymphatic vessels by hyaluronan-mediated docking to the endothelial hyaluronan receptor LYVE-1" Johnson LA. Banerji S Lawrance W, Gileadi, U, Prota, G. Holder, K, Roshorm Y, Hanke T, Cerundolo, V, Gale, N & Jackson DG (2017) Nat. Immunol. 18, 762-770. Multidisciplinary |
Start Year | 2016 |
Description | Robert Gilbert collaboration |
Organisation | University of Oxford |
Department | Wellcome Trust Centre for Human Genetics |
Country | United Kingdom |
Sector | Charity/Non Profit |
PI Contribution | Research using X-Ray crystallography carried out by my Unit postDOC Suneale Banerji with assistance from Robert Gilbert at Wellcome Trust Centre for Human Genomics, Oxford |
Collaborator Contribution | Expertise in Protein Crystallography and associated physical techniques |
Impact | Determination of 3D structure of LYVE-1 alone and in complex with its ligand hyaluronan. Insight into mechanisms by which immune cells use HA and LYVE-1 for interaction with lymphatics during trafficking to LNs in inflammation and immunity. Potential target for therapeutic intervention to prevent inflammation and tissue rejection. Publication in preparation "The lymphatic vessel endothelial hyaluronan receptor LYVE-1 mediates adhesion via unusual sliding interactions with its polymer ligand" Banerji, Bano, Ni, Paci, Lepsik, De Angelis, Gilbert, Richter and Jackson for submission Summer 2022 Multidisciplinary |
Start Year | 2016 |
Description | Role of hyaluronan glycocalyx in T cell migration via lymph |
Organisation | ETH Zurich |
Department | Institute of Pharmaceutical Sciences |
Country | Switzerland |
Sector | Academic/University |
PI Contribution | Advice on experimental design and execution. Reagents. |
Collaborator Contribution | Intravital microscopy, Mouse models |
Impact | Newly started collaboration awaiting outcomes |
Start Year | 2021 |
Description | Sriskandan collaboration |
Organisation | Imperial College London |
Department | Department of Medicine |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Design of experiments and advise on performing experiments to assess interactions between hyaluronan encapsulated Group A strep and lymphatic endothelial cells leading to possible transport to lymph nodes |
Collaborator Contribution | Provision of wild-type and mutant strains of Strep pyogenes and participation of post-doctoral scientist |
Impact | Publication in PLoS Pathogens "Rapid Lymphatic dissemination of encapsulated group A streptococci via lymphatic vessel endothelial receptor-1 interaction" Lynskey, N. Banerji, S, Johnson, LA, Jackson, DG. & Sriskandan, S. (2015) PLoS Pathogens 11, Publication in Nature Communications "Extracellular Bacterial Lymphatic Metastasis Drives Systemic Infection" Siggins MK, Lynskey NN, Lamb LE, Johnson LA, Huse KK, Pearson M, Banerji S, Turner CE, Woollard K, Jackson DG & Sriskandan S. (2020) Nat. Comms. 12 Multidsciplinary Immunology Bacteriology Infectious Diseases |
Start Year | 2012 |
Description | Tony Day collaboration |
Organisation | Wellcome Trust |
Department | Wellcome Trust Centre for Cell-Matrix Research |
Country | United Kingdom |
Sector | Charity/Non Profit |
PI Contribution | Protein expression and purification, Protein structural analysis, intellectual input, co-authoring |
Collaborator Contribution | Expertise in Physical techniques including NMR spectroscopy calorimetry, small angle X ray scattering. Production and purification of hyaluronan oligosaccharides and analogues for binding studies and co-crystallisation with receptors. |
Impact | Ongoing intellectual exchange. Generation of key publications "Structures of the CD44-hyaluronan complex provide insight into a fundamental carbohydrate-protein interaction" Banerji, S, Wright, AJ, Noble, M, Mahoney, DJ, Campbell, ID, Day, AJ & Jackson, DG (2007) Nat. Struct. Mol. Biol. 14, 234-239. "Analysis of CD44-hyaluronan interactions in an artificial membrane system, insights into the distinct binding properties of high and low molecular weight hyaluronan" (2010) Wolny PM, Banerji S, Gounou C, Brisson AR, Day AJ, Jackson DG, & Richter RP. J. Biol. Chem. 285, 30170-30180. "Binding of hyaluronan to the native lymphatic vessel endothelial receptor LYVE-1 is critically dependent on surface clustering and hyaluronan organisation" Lawrance, W, Banerji, S, Day, AJ, Bhattacharjee, S & Jackson, DG (2016) J. Biol. Chem. 291, 8014-8030. "Homodimerisation of the lymphatic endothelial receptor LYVE-1 through a Redox Labile Disulfide is critical for Hyaluronan Binding in Lymphatic Endothelium" Banerji, S, Lawrance, W, Metcalfe, C, Briggs, DC, Yamauchi, A, Dushek, O, van der Merwe, PA, Day, AJ & Jackson, DG (2016) J. Biol. Chem. 291, 25004-25018. |
Start Year | 2016 |
Description | Hands on Participatory event at Swindon Science Festival |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Schools |
Results and Impact | Hands on Participatory event to explain the workings of lymph nodes and the lymphatic system for children to learn by play |
Year(s) Of Engagement Activity | 2020 |
URL | https://www.steam-museum.org.uk/events/event/36/tomorrow_-_swindons_science_festival |
Description | School visit (Oxford) |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | Local |
Primary Audience | Schools |
Results and Impact | Approx 30 primary school pupils participated in an interactive session with Suneal Banerji my senior post-doc who gave them an introductory presentation on immunology and the lymphatic system. This was extremely well received and sparked many questions and discussion both during and afterwards. |
Year(s) Of Engagement Activity | 2016 |