Role of memory B cell migration through the lymph node subcapsular sinus
Lead Research Organisation:
University of Birmingham
Department Name: Institute of Immunology & Immunotherapy
Abstract
Vaccination programmes are a core element of public health strategy worldwide and are key to preventing a wide range of life threatening infections. How vaccines induce immune memory to bacteria and viruses are still poorly understood. We know that vaccination and infection induces B cells to mutate and adapt their genes coding for antibody specific to structures seen on the vaccine. These mutations then will lead to the production of highly specific antibody that can neutralize bacteria or viruses. During this process are also long-lived memory B cells generated, which preserve these mutations for a long time.
We recently observed that memory B cells not only spread through the body and preserve information on vaccines encountered, they also seem to harvest vaccine particles they encounter and transport them back to the places where B cells mature.
The purpose of this project is to understand why memory B cells do this antigen harvest, and which factors regulate it. To understand the regulation, we will produce B cells that cannot produce a range of receptors that direct memory B cell migration. To understand the function of antigen transport, we will disrupt the process and test whether efficient B cell maturation is dependent on continued antigen harvest and transport by memory B cells.
Aged people have problems efficiently producing good antibody responses to vaccines. Some of the defects seen during their B cell maturation may be due to a defect in antigen harvest by memory B cells, and we plan to test whether this process is disrupted in the aged immune system.
Results from this project will help to better understand how vaccines induce protective immune responses and memory. We may better understand how vaccines should be designed or delivered. Better understanding why elderly do not react efficiently to vaccination may lead to ideas on how to correct this process.
We recently observed that memory B cells not only spread through the body and preserve information on vaccines encountered, they also seem to harvest vaccine particles they encounter and transport them back to the places where B cells mature.
The purpose of this project is to understand why memory B cells do this antigen harvest, and which factors regulate it. To understand the regulation, we will produce B cells that cannot produce a range of receptors that direct memory B cell migration. To understand the function of antigen transport, we will disrupt the process and test whether efficient B cell maturation is dependent on continued antigen harvest and transport by memory B cells.
Aged people have problems efficiently producing good antibody responses to vaccines. Some of the defects seen during their B cell maturation may be due to a defect in antigen harvest by memory B cells, and we plan to test whether this process is disrupted in the aged immune system.
Results from this project will help to better understand how vaccines induce protective immune responses and memory. We may better understand how vaccines should be designed or delivered. Better understanding why elderly do not react efficiently to vaccination may lead to ideas on how to correct this process.
Technical Summary
The relevance of migration of germinal centre (GC)-derived memory B cells (mBC) entering the lymph node (LN) subcapsular sinus (SCS), interaction with macrophages and reentry into the GC is tested. Five migratory receptors (CCR7, CXCR3, CCR6, EBI2, S1PR1) are induced during at this stage. We have shown that LN reentry is directed by CCR7.
Objectives:
1) Identify roles of migratory receptors: Cg1Cre mTmG mice, where GC-derived mBC are GFP positive will be used. Cg1Cre mTmG CCR6ko mice are already available and used for CRISP-mediated deletion to produce CCR6 CXCR3 double ko mice. CXCR3 single ko mice will be generated by backcrossing onto wt (Cxcr3 is X-chromosomal). Cg1Cre-EBI2-floxed mice will be generated from available ES cells. S1PR1 will be blocked chemically, which allows study of blockage of combinations of migratory receptors.
Models will be used to study effects on mBC entry into the SCS, entry into other lymphoid and non-lymphoid tissues, and effects on antigen rechallenge.
2) Why is mBC return into the GC is happening? Four hypotheses will be tested:
I. mBC reentry replaces antigen consumed in the GC and sustains the GC while new antigen is produced at the site of entry.
II. unstable membrane bound antigens need to be continuously replaced to support continued GC B cell selection against native undenatured versions of the antigen
III. continued supply of antigen safeguards against escape mutants and supports selection towards antigenic variations
IV. interaction with SCS mBC represents an additional selection step during mBC generation
This will be done using mice generated earlier, by deleting SCS macrophages, or by using Cg1Cre mTmG Ackr4ko mice (available, and deficient in CCR7-dependent mBC reentry).
3) Aged lymph nodes show defects in immune complex deposition in the GC and CCR7 dependent migration. We will test whether reduced vaccine-induced antibody responses are related to reduced SCS mBC - macrophage interaction and mBC reentry.
Objectives:
1) Identify roles of migratory receptors: Cg1Cre mTmG mice, where GC-derived mBC are GFP positive will be used. Cg1Cre mTmG CCR6ko mice are already available and used for CRISP-mediated deletion to produce CCR6 CXCR3 double ko mice. CXCR3 single ko mice will be generated by backcrossing onto wt (Cxcr3 is X-chromosomal). Cg1Cre-EBI2-floxed mice will be generated from available ES cells. S1PR1 will be blocked chemically, which allows study of blockage of combinations of migratory receptors.
Models will be used to study effects on mBC entry into the SCS, entry into other lymphoid and non-lymphoid tissues, and effects on antigen rechallenge.
2) Why is mBC return into the GC is happening? Four hypotheses will be tested:
I. mBC reentry replaces antigen consumed in the GC and sustains the GC while new antigen is produced at the site of entry.
II. unstable membrane bound antigens need to be continuously replaced to support continued GC B cell selection against native undenatured versions of the antigen
III. continued supply of antigen safeguards against escape mutants and supports selection towards antigenic variations
IV. interaction with SCS mBC represents an additional selection step during mBC generation
This will be done using mice generated earlier, by deleting SCS macrophages, or by using Cg1Cre mTmG Ackr4ko mice (available, and deficient in CCR7-dependent mBC reentry).
3) Aged lymph nodes show defects in immune complex deposition in the GC and CCR7 dependent migration. We will test whether reduced vaccine-induced antibody responses are related to reduced SCS mBC - macrophage interaction and mBC reentry.
Planned Impact
There is a broad range of potential beneficiaries of the research that can be summarised as follows:
The scientific community.
The project will produce fundamental new understanding on how vaccine or pathogen antigens are delivered to the place where they induce affinity maturation and the generation of antibody neutralizing pathogens and preventing infection. Understanding these fundamental processes may benefit vaccinology and our understanding how antigens should be delivered and presented in order to produce efficient vaccines that induce antibody that targets the relevant structures on pathogen antigens. Understanding how memory is generated is relevant many different immunological disciplines as it is relevant for T cell immunology, memory, inflammation, autoimmunity, and ageing.
Society
The importance of this work extends significantly beyond supporting academic research. By understanding how antibody responses develop we are helping to understand how to improve vaccine development. This is vital. In our highly mobile society the risks from infectious diseases are increasing due to increased travel and altered global migration and extend also into the food-chain. This is because many of our infections are actively acquired from food (e.g. Salmonella) or livestock rearing acts as an incubator and enables pathogen diversity (e.g. influenza virus). In parallel, we have a decreasing efficacy of anti-microbial treatments due to resistance and only a modest number of anti-virals available. Vaccination is a cost-effective approach that can protect against infection at the extremes of age in those groups that are most susceptible. Furthermore, vaccination is an acceptable intervention to society at large, and as the media response to the recent measles outbreak demonstrates, is one that is diminishing in controversy. Indeed, the measles outbreak demonstrates the importance of vaccination programmes to protecting society and the consequences when there is insufficient vaccine coverage. Theoretical background how vaccines work, what signals regulate the emergence of high affinity B cells and antibody producing cells, and how immunological memory cells are regulated, is still in its infancy.
Industry
The scientific community.
The project will produce fundamental new understanding on how vaccine or pathogen antigens are delivered to the place where they induce affinity maturation and the generation of antibody neutralizing pathogens and preventing infection. Understanding these fundamental processes may benefit vaccinology and our understanding how antigens should be delivered and presented in order to produce efficient vaccines that induce antibody that targets the relevant structures on pathogen antigens. Understanding how memory is generated is relevant many different immunological disciplines as it is relevant for T cell immunology, memory, inflammation, autoimmunity, and ageing.
Society
The importance of this work extends significantly beyond supporting academic research. By understanding how antibody responses develop we are helping to understand how to improve vaccine development. This is vital. In our highly mobile society the risks from infectious diseases are increasing due to increased travel and altered global migration and extend also into the food-chain. This is because many of our infections are actively acquired from food (e.g. Salmonella) or livestock rearing acts as an incubator and enables pathogen diversity (e.g. influenza virus). In parallel, we have a decreasing efficacy of anti-microbial treatments due to resistance and only a modest number of anti-virals available. Vaccination is a cost-effective approach that can protect against infection at the extremes of age in those groups that are most susceptible. Furthermore, vaccination is an acceptable intervention to society at large, and as the media response to the recent measles outbreak demonstrates, is one that is diminishing in controversy. Indeed, the measles outbreak demonstrates the importance of vaccination programmes to protecting society and the consequences when there is insufficient vaccine coverage. Theoretical background how vaccines work, what signals regulate the emergence of high affinity B cells and antibody producing cells, and how immunological memory cells are regulated, is still in its infancy.
Industry
Organisations
- University of Birmingham (Lead Research Organisation)
- Francis Crick Institute (Collaboration)
- AstraZeneca (Collaboration)
- University of Ulm (Collaboration)
- RIKEN (Collaboration)
- University of Basel (Collaboration)
- Leibniz Association (Collaboration)
- UNIVERSITY OF CAMBRIDGE (Collaboration)
- Alloy Therapeutics (Collaboration)
- Helmholtz Association of German Research Centres (Collaboration)
Publications
Roco JA
(2019)
Class-Switch Recombination Occurs Infrequently in Germinal Centers.
in Immunity
Yam-Puc JC
(2021)
Enhanced BCR signaling inflicts early plasmablast and germinal center B cell death.
in iScience
Zhang Y
(2022)
Germinal center derived B cell memory without T cells.
in The Journal of experimental medicine
Jennings E
(2020)
Nr4a1 and Nr4a3 Reporter Mice Are Differentially Sensitive to T Cell Receptor Signal Strength and Duration.
in Cell reports
Darby M
(2019)
Pre-conception maternal helminth infection transfers via nursing long-lasting cellular immunity against helminths to offspring
in Science Advances
Zhang Y
(2022)
Recycling of memory B cells between germinal center and lymph node subcapsular sinus supports affinity maturation to antigenic drift.
in Nature communications
Yam-Puc J
(2018)
Role of B-cell receptors for B-cell development and antigen-induced differentiation
in F1000Research
Description | We found how immune memory of B cell spreads through the body, and which factors affect the migration of B cells during this process. While doing this we found a new way B cells interact with mutating pathogens. This may have important implications for our understanding of B cell memory and adaption to antigenic drift, as it happens for mutating pathogen, e.g. HIV, influenza, Covid-19, or Dengue |
Exploitation Route | Understanding and design of vaccines. Understanding of the immune response to pathogens may help treating long term or recurrent illness to mutating pathogens. |
Sectors | Pharmaceuticals and Medical Biotechnology |
Title | Recycling of memory B cell supports affinity maturation to antigenic drift |
Description | Gene expression data for memory B cells and germinal centre cells |
Type Of Material | Database/Collection of data |
Year Produced | 2022 |
Provided To Others? | Yes |
Impact | Led to manuscript "Recycling of memory B cells between germinal center and lymph node subcapsular sinus supports affinity maturation to antigenic drift" currently under review |
URL | https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE188687 |
Title | Recycling of memory B cells between germinal center and lymph node subcapsular sinus supports affinity maturation to antigenic drift |
Description | ImageJ macro for automatic subcapsular sinus segmentation |
Type Of Material | Computer model/algorithm |
Year Produced | 2022 |
Provided To Others? | Yes |
Impact | Led to publication "Recycling of memory B cells between germinal center and lymph node subcapsular sinus supports affinity maturation to antigenic drift" currently under review |
URL | https://github.com/JeremyPike/SCS-cell-tracking |
Title | Recycling of memory B cells supports affinity maturation to antigenic drift |
Description | Customised Fiji code for intravital cell tracking |
Type Of Material | Computer model/algorithm |
Year Produced | 2022 |
Provided To Others? | Yes |
Impact | Led to publication "Recycling of memory B cell between germinal center and lymph node subcapsular sinus supports affinity maturation to antigenic drift", currently under review. |
URL | https://github.com/JeremyPike/SCS-cell-tracking |
Description | An anergic B cell model for antibody discovery |
Organisation | Alloy Therapeutics |
Country | United States |
Sector | Private |
PI Contribution | We developed and characterised a new mouse strain (IgMg1 mouse) with hyperactive B cell receptor signalling, leading to general B cell anergy. We plan to test whether this model is suitable for antibody discovery. |
Collaborator Contribution | Hosting an BBSRC iCASE student to test the model using antigens used in industry, and access to lab facilities, e.g. 10x genomics. |
Impact | None yet. Industry collaboration. |
Start Year | 2020 |
Description | Gene expression in memory B cells |
Organisation | Francis Crick Institute |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Provision of mRNA from in vivo differentiated memory B cells |
Collaborator Contribution | Gene expression analysis from different in vivo differentiated memory B cells by RNAseq and bioinformatic analysis |
Impact | New data on gene expression in new memory B cell subsets. |
Start Year | 2015 |
Description | In silico modelling of germinal centres |
Organisation | Helmholtz Association of German Research Centres |
Department | Helmholtz Centre for Infection Research (HZI) |
Country | Germany |
Sector | Academic/University |
PI Contribution | Data provision for in silico models Work on the in silico model |
Collaborator Contribution | In silico modelling Predictions for optimal experimental in vivo setups |
Impact | The collaboration has led to several publications. It has produced a new in silico model to analyse the regulation of affinity maturation in germinal centres. This has lead to several further collaborations and publications for our collaborators. |
Start Year | 2006 |
Description | Industrial partnership |
Organisation | AstraZeneca |
Department | MedImmune |
Country | United Kingdom |
Sector | Private |
PI Contribution | We are analysing GM mouse strains with altered B cell receptor signalling in germinal centres for their suitability as tools to produce new therapeutic monoclonal antibody drugs. The prediction is that the GM mice will easier produce specific high affinity antibodies to difficult target antigens. Further, we are analysing effects of these genetic modification on the regulation of high affinity antibody responses. |
Collaborator Contribution | The industrial partners design and produce the initial vectors for genetic manipulation of mice. They transfect these into embryonic stem cells and produce GM mice. After this step the mice are transferred to us. At a later stage they will test the mice for practical usability as vehicles to produce high affinity B cells to difficult targets for hybridoma fusions. They will use the mice and test whether they ca produce monoclonal antibodies using some of there identified human therapeutic targets. |
Impact | Three GM mouse strains have been produced and are currently analysed in our lab, publications in preparation. New iCASE PhD studentship to work on 4th strain. |
Start Year | 2014 |
Description | Intravital imaging of memory B cell migration |
Organisation | University of Cambridge |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Provision of GMO mice, |
Collaborator Contribution | Immunisation of GMO mice and intravital imaging of migrating memory B cells. Training visits for UoB staff on intravital microscopy in preparation. |
Impact | Early results show new patterns of memory B cell migration and roles of chemokine receptors directing this. |
Start Year | 2016 |
Description | Intravital imaging of plasma cell differentiation |
Organisation | Leibniz Association |
Department | German Rheumatism Research Centre |
Country | Germany |
Sector | Charity/Non Profit |
PI Contribution | A postdoctoral research fellow spent twice 2 months at DRFZ to learn intravital imaging techniques and study plasma cell differentiation from germinal centres |
Collaborator Contribution | Hosting, training, and help with experiments. Provision of experimental animals |
Impact | Training of postdoctoral research fellow Preliminary data on plasma cell differentiation Submission of funding proposal to Wellcome Trust for an intravital microscope |
Start Year | 2011 |
Description | Memory B cell migration and activation of secondary responses to antigenic variation |
Organisation | University of Basel |
Department | Department of Biomedicine |
Country | Switzerland |
Sector | Academic/University |
PI Contribution | Advise and provision of virus vectors that can be used to study variable virus antigens |
Collaborator Contribution | The partner plans to provide virus samples that can be used at UoB for immunisation experiments |
Impact | none yet |
Start Year | 2018 |
Description | Provision of GM mice |
Organisation | RIKEN |
Department | RIKEN Center for Integrative Medical Sciences (IMS) |
Country | Japan |
Sector | Private |
PI Contribution | Provision of mice that can be used to track germinal centre B cells |
Collaborator Contribution | Provision of mice that have an inducible expression of Cre recombinase under the control of the S1PR2 promotor |
Impact | We currently use these mice to generate new unique mouse models to study germinal centre B cell differentiation. |
Start Year | 2018 |
Description | Provision of mice with anergic B cell |
Organisation | University of Ulm |
Country | Germany |
Sector | Academic/University |
PI Contribution | This is to support our ongoing collaboration with University Freiburg, Kathrin Klaesner, Michael Reth to study B cell receptor signalling the anergic IgMg1 anergic B cell mouse, that was generated and characterised by us. |
Collaborator Contribution | Transgenic mice with anergic B cells (HEL-anti HEL system) are provided as controls for ongoing collaboration with University Freiburg, Kathrin Klaesner, Michael Reth, to study B cell receptor signalling in our IgMg1 anergic B cells |
Impact | Data on the role for B cell receptor signalling for anergy and B cell tolerance |
Start Year | 2020 |