The biology of lymphatics in inflammation, immunity and cancer
Lead Research Organisation:
University of Oxford
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.
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, United Kingdom (Collaboration, Lead Research Organisation)
- Centre for Cooperative Research in Biomaterials (CIC BiomaGUNE) (Collaboration)
- Swiss Federal Institute of Technology in Lausanne (EPFL) (Collaboration)
- University of California, San Francisco, United States (Collaboration)
- Imperial College London, United Kingdom (Collaboration)
- University Clinic Cologne (Collaboration)
- Oxford University Hospitals NHS Foundation Trust, Headington (Collaboration)
- University of Houston, United States (Collaboration)
- University of Birmingham, United Kingdom (Collaboration)
- Medical Research Council (Collaboration)
- Regeneron Pharmaceuticals, Inc. (Collaboration)
- Medical University of Vienna (Collaboration)
- University of Seville, Spain (Collaboration)
- Cardiff University, United Kingdom (Collaboration)
- Wellcome Trust, LONDON (Collaboration)
Publications

Banerji S
(2016)
Homodimerization of the Lymph Vessel Endothelial Receptor LYVE-1 through a Redox-labile Disulfide Is Critical for Hyaluronan Binding in Lymphatic Endothelium.
in The Journal of biological chemistry

Bano F
(2016)
A single molecule assay to probe monovalent and multivalent bonds between hyaluronan and its key leukocyte receptor CD44 under force.
in Scientific reports

Bauer J
(2018)
TGFß counteracts LYVE-1-mediated induction of lymphangiogenesis by small hyaluronan oligosaccharides.
in Journal of molecular medicine (Berlin, Germany)

Brown M
(2018)
Lymphatic exosomes promote dendritic cell migration along guidance cues.
in The Journal of cell biology

Chen X
(2016)
Micromechanical Analysis of the Hyaluronan-Rich Matrix Surrounding the Oocyte Reveals a Uniquely Soft and Elastic Composition.
in Biophysical journal


Jackson DG
(2014)
Lymphatic Regulation of Cellular Trafficking.
in Journal of clinical & cellular immunology

Jackson DG
(2019)
Leucocyte Trafficking via the Lymphatic Vasculature- Mechanisms and Consequences.
in Frontiers in immunology

Jackson DG
(2019)
Hyaluronan in the lymphatics: The key role of the hyaluronan receptor LYVE-1 in leucocyte trafficking.
in Matrix biology : journal of the International Society for Matrix Biology

Johnson LA
(2017)
Dendritic cells enter lymph vessels by hyaluronan-mediated docking to the endothelial receptor LYVE-1.
in Nature immunology
Description | Academy of Finland |
Geographic Reach | Europe |
Policy Influence Type | Participation in a advisory committee |
Impact | Advised on appointment of prestigious Professorship |
Description | Finnish Academy of Sciences |
Geographic Reach | Europe |
Policy Influence Type | Influenced training of practitioners or researchers |
Impact | Sat on Grant review committee |
Description | Fondazione Cariplo grant review |
Geographic Reach | Europe |
Policy Influence Type | Participation in a advisory committee |
Impact | Reviewed grant proposals for Italian biomedical science funding body |
Description | Lund University |
Geographic Reach | Asia |
Policy Influence Type | Participation in advisory committee |
Impact | Advised on appointment of prestigious Professorship |
Description | Membership of MRC Infection and Immunity Board |
Geographic Reach | National |
Policy Influence Type | Participation in advisory committee |
Description | Organisation of University postgraduate research training course |
Geographic Reach | Local/Municipal/Regional |
Policy Influence Type | Influenced training of practitioners or researchers |
Impact | Optimisation of Institute lecture course for first year DPhil students with aim of broadening knowledge in key areas of biomedical research techniques |
Description | Review of DK Plus postgraduate research training program |
Geographic Reach | Europe |
Policy Influence Type | Influenced training of practitioners or researchers |
Impact | Evaluation and rating of PhD students and PIs associated with Austrian Government funded program |
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 | 04/2014 |
End | 03/2017 |
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 |
Title | Crystal structure of CD44 receptor ligand complex |
Description | First crystal showing molecular details of interaction between a hyaluronan receptor and its ligand hyaluronan. Determined structure in the case of the primary receptor CD44 that is involved in inflammatory leukocyte trafficking and tumour metastasis |
Type Of Material | Technology assay or reagent |
Provided To Others? | No |
Impact | Major boost to fundamental biology. Template for design of small molecule inhibitors of CD44 function with possible therapeutic potential |
Title | Function blocking antibodies |
Description | Defined properties of mAbs generated against lymphatic vessel receptor as being function blocking for leukocyte trafficking in lymphatics |
Type Of Material | Technology assay or reagent |
Provided To Others? | No |
Impact | Use of mAbs identifies lymphatic receptor as important for leukocyte movement from tissues to lymph nodes in inflammation |
Title | Primary lymphatic endothelial cells |
Description | Primary lymphatic endothelial cells isolated by immunomagnetic bead selection from human skin, mouse skin and human colorectal tissue |
Type Of Material | Cell line |
Provided To Others? | No |
Impact | Ability to analyze molecular details of lymphatic vessel physiology and to establish various assays for cell adhesion / transmigration studies with cells of authentic phenotype |
Title | Tumour-induced lymphatic vessel markers |
Description | Identified molecular markers induced in lymphatic vessels of murine metastatic tumours by comparison with normal tissue lymphatics. Confirmed expression also in human cancers. |
Type Of Material | Model of mechanisms or symptoms - mammalian in vivo |
Provided To Others? | No |
Impact | Used lymphatics markers to predict metastasis in colorectal/head and neck cancers. Identified receptor molecules that are liely to be mechanistically involved in lymphatic metastasis of tumours. |
Description | Aspp1 collaboration |
Organisation | University of Oxford |
Department | Nuffield Department of Medicine |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Expertise reagents and advice |
Collaborator Contribution | Expertise reagents and advice |
Impact | None as yet |
Start Year | 2016 |
Description | Awen Gallimore collaboration |
Organisation | Cardiff University |
Department | Infection and Immunity |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Advice on experimental design and evaluation of experimental data from mouse tumour model |
Collaborator Contribution | Carried out studies into mouse fibrosarcoma tumour development and role of different vasculatures in T cell recruitment to tumour |
Impact | Manuscript in International Journal of Cancer PMID 24142504 |
Start Year | 2014 |
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 in press in Investigational Ophthalmology and Visual Science 2019 |
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 | David Coleman collaboration |
Organisation | Oxford University Hospitals NHS Foundation Trust |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Regular immunoisolation of primary endothelial cells from human surgically excised tissue - generation of materials essential for our research |
Collaborator Contribution | Provision of human skin samples for isolation of primary cells |
Impact | Regular supply of primary cells vital for our research and fundamental to most publications coming from my group |
Start Year | 2016 |
Description | David Greaves collaboration |
Organisation | University of Oxford |
Department | Sir William Dunn School of Pathology |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Intellectual input and experimental design Animal experiments involving cell trafficking |
Collaborator Contribution | Advice and expertise in measurement of endothelial cell junctional permeability Contribution of transgenic reporter mice |
Impact | Manuscript in Blood; describing utility of CD68 transgenic reporter mice for macrophage trafficking studies Manuscript in preparation for J Biol Chem |
Start Year | 2012 |
Description | Dontscho Kerjaschki |
Organisation | Medical University of Vienna |
Department | Clinical Institute of Pathology |
Country | Austria |
Sector | Academic/University |
PI Contribution | Research collaboration leading to joint publication - currently in final revision at Journal of Cell Biology |
Collaborator Contribution | Carried out experimental work at host Institute and provided us with reagents to perform related studies in Oxford |
Impact | This multidisciplinary collaboration has yielded a publication in a prestigious high-impact journal (Journal of Cell Biology) that is currently in its final round of revision with decision expected imminently |
Start Year | 2017 |
Description | Errin Johnson collaboration |
Organisation | University of Oxford |
Department | Sir William Dunn School of Pathology |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Advice and intellectual contribution |
Collaborator Contribution | Electron microscopy |
Impact | Ongoing |
Start Year | 2015 |
Description | Fluorescence and EM imaging of lung |
Organisation | Medical University of Vienna |
Department | Clinical Institute of Pathology |
Country | Austria |
Sector | Academic/University |
PI Contribution | Provision of reagents and intellectual input |
Collaborator Contribution | Performed immunoelectron microscopy and immunofluorescence imaging of thin sections of mouse alveolae |
Impact | Obtained important data for inclusion in manuscript under preparation for publication in scientific journal |
Start Year | 2012 |
Description | High Resolution confocal Lung Imaging |
Organisation | University of California, San Francisco |
Department | Department of Anatomy |
Country | United States |
Sector | Academic/University |
PI Contribution | Provided reagents and intellectual input |
Collaborator Contribution | Carried out perfusion fixation and inflation of lungs and prepared sections for confocal imaging and z sectioning to identify critical cell population in alveolae |
Impact | Obtained critical data for inclusion in a manuscript under preparation for submission to scientific journal |
Start Year | 2012 |
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 | Kevi Maloy collaboration |
Organisation | University of Oxford |
Department | Sir William Dunn School of Pathology |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Reagents expertise and advice |
Collaborator Contribution | Reagents expertise and advice |
Impact | None as yet |
Start Year | 2016 |
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 | Luxheng Xue collaboration |
Organisation | University of Oxford |
Department | Nuffield Department of Medicine |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Advice and experimental design. Discussion of data and manuscript editing |
Collaborator Contribution | Performance of experiments to investigate neutrophil : T cell crosstalk |
Impact | Manuscript in J allergy and Clinical Immunol PMID 25441644 |
Start Year | 2013 |
Description | Melody Swartz collaboration |
Organisation | Swiss Federal Institute of Technology in Lausanne (EPFL) |
Department | Institute of Bioengineering |
Country | Switzerland |
Sector | Academic/University |
PI Contribution | Travel to partners laboratory for training |
Collaborator Contribution | Training in the immunoisolation of lymphatic endothelial cells from mice using podoplanin mAbs. Training in intravital imaging of lymphatics. |
Impact | Transfer of specialist expertise |
Start Year | 2013 |
Description | Paul Riley collaboration |
Organisation | University of Oxford |
Department | Department of Physiology, Anatomy and Genetics |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Reagents advice and expertise |
Collaborator Contribution | Reagents advice and expertise |
Impact | None as yet |
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 | 27679982 27332136 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 (Johnson et al Nature Immunology In Press 2017 ) Stimulating ongoing research in new areas, identification of possible new targets for anti-inflammatory therapy and future access to further transgenic animals |
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 | Insight into 3D structure of LYVE-1 protein and the mode of its interaction with hyaluronan core to focus of my group research |
Start Year | 2016 |
Description | Sriskandan collaboration |
Organisation | Imperial College London |
Department | Faculty 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 2015 PMID 26352587 Multidsciplinary Immunology Bacteriology Infectious Diseases |
Start Year | 2012 |
Description | Steven Watson collaboration |
Organisation | University of Birmingham |
Department | College of Medical and Dental Sciences |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Expertise advice and reagents |
Collaborator Contribution | Expertise advice and reagents |
Impact | Publication in J Biol Chem PMID 25368330 |
Start Year | 2015 |
Description | Super resolution microscopy |
Organisation | Medical Research Council (MRC) |
Department | MRC Human Immunology Unit |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Preparation of primary cells and transfectants, and appropriate mAbs and conjugates for STED microscopic evaluation of receptor surface distribution |
Collaborator Contribution | Performing STED microscopy and analysis of data |
Impact | Multidisciplinary collaboration Manuscript submitted to Scientific Reports 2016 |
Start Year | 2014 |
Description | Surface Plasmon resonance (Biacore) analysis of receptor ligand binding affinity |
Organisation | University of Oxford |
Department | Sir William Dunn School of Pathology |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Generated recombinant soluble hyaluronan receptors and mutants thereof and performed Biacore analyses using hyaluronan coated chip to determine binding affinities |
Collaborator Contribution | Mathematical evaluation of binding data from kinetic analyses and derivation of accurate Kd values |
Impact | Manuscript that is about to be submitted to J Biol Chem |
Start Year | 2014 |
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 27733683 26823460 and impending new publication describing crystal structure of ligand binding domain of lymphatic hyaluronan receptor LYVE-1 |
Start Year | 2016 |
Title | Licensing of monoclonal antibodies |
Description | Hybridomas producing monoclonal antibodies to human LYVE-1 were licensed to Merck Millipore on a non-exclusive basis.The mAbs will be included in their catalogue for non-medical use. |
IP Reference | |
Protection | Protection not required |
Year Protection Granted | 2016 |
Licensed | Yes |
Impact | N.A. |
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 |