MRC/FAPESP - bilateral agreement: Multiple-epitope vaccine to confer serotype-independent protection against pneumonia

Lead Research Organisation: Liverpool School of Tropical Medicine
Department Name: Clinical Sciences

Abstract

Pneumococcal infections account for 11% of all deaths in children under 5 years of age and in older adults, Streptococcus pneumoniae is the leading cause of community-acquired pneumonia. Pneumococcal infections in both children and adults are potentially preventable by vaccination but current vaccines do not offer the level of protection needed. Furthermore, the use of polysaccharide conjugate vaccine is associated with disease caused by non-vaccine types, a phenomenon known as serotype replacement. The existence of >90 different pneumococcal types implies that serotype replacement will be a persistent problem for anti-capsular vaccines. The development of a vaccine based on pneumococcal proteins common to all strains offers the potential for serotype-independent protection and the complete eradication of carriage.
Pneumococcal surface proteins A (PspA) and Pneumococcal surface protein C (PspC) among the most promising candidates for a protein-based vaccine. Although these proteins are variable among different clinical isolates, we have found some variants that are able to induce antibodies with broad reactivity, recognizing different PspA and PspC variants. We have also shown that fusion of PspA protein fragments is an efficient strategy to extend protection provided by a protein, but the ideal epitopes (parts of the protein) to be included in the vaccine has yet to be defined. The selection of epitopes capable of inducing antibodies with broad cross-reactivity (able to recognize all variants of PspA and PspC) is of clinical importance to the successful development of a vaccine using this approach.
We will use peptide array techniques to describe the entire range of cross-reactive epitopes target by sera that are known to be cross-reactive (murine immunized and human sera) and determine the optimal epitopes to be included in the vaccine. The approach of using peptide arrays has been successful used to identify immunogenic epitopes of candidates for Plasmodium and Leptospira vaccine but not yet for S. pneumoniae.
Our group at LSTM has successful established an Experimental Human Pneumococcal Carriage (EHPC) model and has established pneumonia patient cohorts. We will use a unique set of pre- and post-pneumococcal intranasal inoculation samples to dissect responses to PspA and PspC epitopes to a known carriage event. We will use samples from pneumonia patients to compare healthy and susceptible antibody function. Results obtained in a previous experimental human carriage study showed that presence of serum antibodies against PspA correlated with failure to establish carriage. We have observed increased antibody responses to PspA and PspC in serum, nasal wash and bronchoalveolar lavage following carriage. Immunoglobulin reponses to exposure, carriage and disease are different and show compartmentalization.
There is an urgent need for a pneumonia vaccine that is effective in childhood and in old age. We propose that an optimal vaccine will protect both age groups against pneumonia caused by all serotypes of pneumococci. We will achieve this by identifying epitopes of PspA and PspC to produce a multiple-epitope vaccine, test this vaccine in murine models of invasive disease and confirm cross-reactivity of antibodies using a wide panel of pneumococcal clinical isolates. Further we will use the effective acquired immunity that protects healthy adults against carriage and the deficient response of pneumonia patients to determine the effective levels and function of antibodies to this vaccine candidate. Finally, we will confirm induction of cellular responses elicited by immunisation in mice and the presence of multiple-epitope antigen specific CD4 T cells in healthy adults and pneumonia patients.

Technical Summary

Invasive pneumococcal disease is a leading cause of morbidity and premature mortality around the world. Although current vaccination strategies are effective, they have major limitations such as low levels of protection against pneumonia and serotype replacement.
We have conducted extensive basic laboratory and clinical translational research that has substantially improved our understanding of the limitations of current vaccines and provided insights into new approaches for vaccine development.
We now propose a collaborative project between BI and LSTM that draws on previous successful projects to identify targets for a completely new type of pneumonia vaccine that will confer serotype-independent protection against pneumonia.
Pneumococcal surface protein A (PspA) and Pneumococcal surface protein C (PspC) are among the most promising vaccine candidates but they exhibit antigen variability. We will describe the entire range of PspA and PspC peptides in peptide arrays that will be probed with serum from immunized mice and adults exposed to pneumococci during experimental carriage to identify optimal cross-reacting epitopes to be included in a multiple-epitope vaccine.
We will test the protective potential of this vaccine in murine models and serum cross-reactivity in in vitro assays. We will confirm induction of cellular responses in immunized mice and the presence of multiple-epitope antigen specific CD4 T cells in healthy adults and pneumonia patients after ex-vivo stimulation. We will determine responses to vaccine associated with protection from experimental human pneumococcal carriage and with susceptibility to pneumonia disease.
These experiments will add a novel approach to protein vaccine discovery and knowledge regarding optimal vaccination strategy for mucosal protection.

Planned Impact

The impact of this grant will be mainly on the partner Institutions and the development of a novel collaboration. Our Institutions are of similar age and have committed to serve least served populations. The Butantan Institute (BI) has a long standing expertise in vaccine technology that is novel to Liverpool School of Tropical Medicine (LSTM). LSTM has wide international links and an experimental/ medicine platform that is novel to BI.
1. Impact of the collaboration on partner Institutions
I. Change in the culture of organizations (LSTM and BI). We intend to add vaccine research to LSTM portfolio and improve BI international collaboration and international exposure. Vaccine research has long been the focus of BI effort but is new to the LSTM portfolio.
II. Attract R&D investment: LSTM will help BI to partner with a UK vaccine production company.
BI has expertise in technology transfer for vaccine production. BI will gain from partnership with a UK based vaccine company to transfer technology for production of other vaccines important to Brazil and other least economically developed countries (LEDC).
2. Impact on project staff and students
I. International exposure: BI researchers will gain from enhanced impact publications, international collaborators and conferences. International exposure will bring more international collaborators and joined grant applications.
II. Collaborative training: C Vadesilho (MSc student) and the PhD student that will continue the Multiple-epitope project at BI will have the opportunity of internships in the LSTM laboratory. BI students will gain from opportunity of visiting and work in an international laboratory environment. All LSTM staff will gain from the exchange of knowledge and experience during visit of international students and researchers. Staff will also benefit from future grant applications.
3. Impact on general public
I. Public involvement: General public will gain knowledge that will be disseminated during "Meet the public days" and school visits, which are regular events on both institutions. BI and LSTM have a strong past record of successful public engagement and are committed to improving the public understanding of science.
4. Impact on vulnerable populations
I. LSTM and BI have close collaboration with least economically developed countries (LEDC). The global burden of pneumococcal disease is highest in vulnerable LEDC and a successful vaccine will be targeting particularly those LEDC that do not have a pneumococcal vaccine in their national immunization program. Vulnerable population living in these countries such as woman, children and HIV population will benefit from a new vaccine.
5. Vaccination Policy
I. Vaccine success will impact in change of vaccination policy. BI is affiliated with Ministry of Health in Brazil and both produces vaccines used in the Brazilian vaccine programme and advices in vaccination policy. Our results will directly feed into a body of work that is used to inform government on vaccination strategy.

Publications

10 25 50
 
Description A phase 2, randomized double-blind placebo-controlled study to evaluate the safety, tolerability and efficiacy of GEN-004
Amount £1,180,000 (GBP)
Funding ID 50190 
Organisation Genocea Biosciences 
Sector Private
Country Unknown
Start 08/2014 
End 07/2016
 
Description Effect of Live Attenuated and Inactivated Influenza vaccines on Experimental Human Pneumococcal Carriage
Amount $2,518,294 (USD)
Funding ID OPP1117728 
Organisation Bill and Melinda Gates Foundation 
Sector Charity/Non Profit
Country United States
Start 11/2014 
End 06/2017
 
Description Enhancing mucosal immunity to Streptococcus pneumoniae by nasal administration of live strains attenuated in virulence
Amount £523,377 (GBP)
Funding ID MR/N02687X/1 
Organisation Medical Research Council (MRC) 
Sector Academic/University
Country United Kingdom
Start 05/2016 
End 04/2019
 
Description Experimental Human Pneumococcal Carriage to determine optimal protection from carriage and mechanisms of mucosal immunisation against disease
Amount £2,282,780 (GBP)
Funding ID MR/M011569/1 
Organisation Medical Research Council (MRC) 
Sector Academic/University
Country United Kingdom
Start 11/2014 
End 10/2019
 
Description Jean Clayton Travel Award - Jessica Owugha
Amount £2,470 (GBP)
Organisation Liverpool School of Tropical Medicine 
Sector Academic/University
Country United Kingdom
Start 10/2014 
End 06/2015
 
Description LSTM PhD award of Jessica Owugha
Amount £41,178 (GBP)
Funding ID LSTM 200499290 
Organisation Liverpool School of Tropical Medicine 
Sector Academic/University
Country United Kingdom
Start 10/2013 
End 09/2016
 
Description MRC Proximity to Discovery funds
Amount £20,000 (GBP)
Organisation Medical Research Council (MRC) 
Sector Academic/University
Country United Kingdom
Start 10/2017 
End 08/2018
 
Description Studentship Cintia Vadesilho for peptide array discovery at LSTM
Amount £4,460 (GBP)
Organisation São Paulo Research Foundation (FAPESP) 
Sector Public
Country Brazil
Start 04/2013 
End 06/2013
 
Description Vaccine Fapesp MICA : Pulmonary Delivery of a Targeted Mucosal Nanocarrier Vaccine for Pneumonia
Amount £456,762 (GBP)
Funding ID MR/P022758/1 
Organisation Medical Research Council (MRC) 
Sector Academic/University
Country United Kingdom
Start 02/2017 
End 02/2020
 
Description Bilateral Partnership LSTM/BI 
Organisation Butantan Institute
Department Biotechnology Centre
Country Brazil 
Sector Public 
PI Contribution We have received visiting students from Butantan Institute. Two students (Adriana Moreno and Cintia Vadesilho) have visited our laboratory at LSTM for 3 months each. They gained from the experience of being part of a research laboratory team during their PhD and MSc studies.
Collaborator Contribution Butantan Institute has provided important advice on pneumococcal protein discovery as well as supplied LSTM lab with several recombinant proteins and DNA plasmids for protein expression.
Impact Internship of Cintia Vadesilho LSTM PhD award to Jessica Owugha Eliane Miyaji's visit at LSTM Daniela Ferreira's visit at BI
Start Year 2009
 
Description Collaboration with Liverpool John Moores University - Nanoparticles for pulmonary delivery of vaccines 
Organisation Liverpool John Moores University
Country United Kingdom 
Sector Academic/University 
PI Contribution We have been working together with the team at LJMU to optimize nanoparticles for the delivery of PspA antigens. We have provide laboratory support for this project as well as clinical samples.
Collaborator Contribution The team at LJMU have performed characterisation of particles loaded with PspA
Impact Three publications have already resulted from this partnership and a new grant from the Medical Research Council/FAPESP awarded to Dr Imran Saleem in 2017
Start Year 2014
 
Description Collaboration with University of Sao Paulo (Helder Nakaya) for bioinformatic analysis 
Organisation University of Sao Paulo
Country Brazil 
Sector Academic/University 
PI Contribution Our team has provided immunology data obtained during EHPC studies
Collaborator Contribution The team at University of Sao Paulo has performed computacional analysis to identify differential expression between volunteers protected vs non protected from pneumococcus.
Impact An award to Helder Nakaya from the University of Sao Paulo to visit LSTM in 2017 (awarded in 2016) . The data from this collaboration was used for 2 grant applications (one will be submitted to the MRC in May 2017).
Start Year 2013
 
Description Computational approaches to common conserved epitope to 30 antigens 
Organisation University of Oxford
Department Jenner Institute
Country United Kingdom 
Sector Academic/University 
PI Contribution Ongoing collaboration with Dr Arturo Reyes-Sandoval
Collaborator Contribution TBC
Impact TBC
Start Year 2013
 
Description Science without Borders 
Organisation Liverpool School of Tropical Medicine
Country United Kingdom 
Sector Academic/University 
PI Contribution Launched LSTM PhD SwB Programme and went to Brazil on a tour of Universities to attract bright PhD students to LSTM. 3 Brazilian students have applied to the programme this year.
Collaborator Contribution TBC
Impact tbc
Start Year 2014
 
Description Science without Borders 
Organisation State University of Campinas
Country Brazil 
Sector Academic/University 
PI Contribution Launched LSTM PhD SwB Programme and went to Brazil on a tour of Universities to attract bright PhD students to LSTM. 3 Brazilian students have applied to the programme this year.
Collaborator Contribution TBC
Impact tbc
Start Year 2014
 
Description Science without Borders 
Organisation University of Sao Paulo
Country Brazil 
Sector Academic/University 
PI Contribution Launched LSTM PhD SwB Programme and went to Brazil on a tour of Universities to attract bright PhD students to LSTM. 3 Brazilian students have applied to the programme this year.
Collaborator Contribution TBC
Impact tbc
Start Year 2014
 
Description UK influenza research networks 
Organisation Department of Health (DH)
Department Human Tissue Authority (HTA)
Country United Kingdom 
Sector Public 
PI Contribution During the H1N1 pandemic, Liverpool recruited as two of four sites for the MOSAIC consortium. Subsequently, we are poised as partners in the HTA funded ASAP study to study the effect of steroids should pandemic influenza recur. Partnerships led by Prof P Openshaw and Dr WS Lim.
Collaborator Contribution Major recruiting site for MOSAIC study (WT/MRC) and planned key site for ASAP study (NIHR HTA).
Impact Publications, planned new studies.
Start Year 2011
 
Title GEN-004 is a recombinant Streptococcus pneumoniae protein subunit vaccine 
Description GEN-004 is a recombinant Streptococcus pneumoniae protein subunit vaccine consisting of 3 recombinant antigens. Aluminium hydroxide is used as an adjuvant. In a Phase 1 study, GEN-004 met its safety, tolerability and immunogenicity goals, including measurable increases in the blood of TH17 cells. The randomized, double-blind, dose-escalation, placebo-controlled clinical trial included approximately 90 healthy adult volunteers. 
Type Therapeutic Intervention - Vaccines
Year Development Stage Completed 2014
Development Status Under active development/distribution
Clinical Trial? Yes
Impact tbc 
 
Description 9th International Symposium on Pneumococci and Pneumococcal Diseases, ISPPD-9, India 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Talk to disseminate research findings and discussion of future collaborative activities

tbc
Year(s) Of Engagement Activity 2014
 
Description A visit to LSTM from Helder Nakaya 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Professional Practitioners
Results and Impact Helder Nakaya visited LSTM to discuss the analysis for the RNA-Seq for the LAIV1 study, and to discuss the integrative analysis of other samples collected.
Year(s) Of Engagement Activity 2017
 
Description Academic visit and Talk at Butantan Institute and University of Sao Paulo. 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Postgraduate students
Results and Impact Postgraduate, undergraduate students and principal investigators attended these two seminars which sparked questions and discussion afterwards. This visit initiated the visit of two academics from Brazil to the LSTM in 2017 (Helder Nakaya and Alessandra Schanoski).
Year(s) Of Engagement Activity 2016
 
Description Academic visit to Butantan Institute and Univeristy of Sao Paulo 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Other audiences
Results and Impact Daniela Ferreira visited the University of Sao Paulo and the Butantan Institute for discussions with Eliane Miyaji, Helder Nakaya and Maria Leonor regarding Jessica Owugha's publication - MRC/FAPESP - bilateral agreement: Multiple-epitope vaccine to confer serotype-independent protection against pneumonia. The collaborators also discussed the Pulmonary Delivery of a Targeted Mucosal Nanocarrier Vaccine for Pneumonia project, which is a follow on piece of work as a result of the FAPESP project.
Year(s) Of Engagement Activity 2017
 
Description Annually Invited Lecture at Butantan Institute, Sao Paulo, Brazil 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact As part of the collaboration between LSTM and Butantan Institute underpinned by the MRC/FAPESP and MRC Programme grant awards DF visits Butantan annually and give a talk about project results aimed at all researchers and students at the Institute

tbc
Year(s) Of Engagement Activity 2010,2012,2013,2014,2015
 
Description BREATHE theme leaders visit Malawi CAPS sites 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Participants in your research and patient groups
Results and Impact We planned exposure monitoring for 10000 children, and wrote 2 new grants.

New grants including MRC NIRG and NIH RO1
Year(s) Of Engagement Activity 2013
URL http://www.capstudy.org/
 
Description Biological Challenges of effective vaccines - Royal Society, London 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Type Of Presentation keynote/invited speaker
Geographic Reach International
Primary Audience Supporters
Results and Impact This satellite meeting discussed recent findings and challenge participants to identify key research questions that will yield more effective vaccines in low-income countries.

My talk was discussed on twitter by Pathogens
Year(s) Of Engagement Activity 2015
 
Description Daniela Ferreira - visit and talk at BI 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact Daniela Ferreira visited Butantan Institute (BI- (Sao Paulo, Brazil) on the 17th and 18th of October 2013. Daniela met with collaborators Paulo Lee Ho, Maria Leonor Oliveira and Eliane Miyaji and gave a talk on "Experimental Human Pneumococcal Carriage for vaccine research" that was attended by 30 researchers and students.

During this visit it was discussed a possible laboratory internship of Jessica Owugha (PhD LSTM) to Butantan Institute and further grant applications relating to the Me vaccine project (MR/K01188X/1).
Year(s) Of Engagement Activity 2013
 
Description Dr Eliane Miyaji (Butantan Institute) Visit to LSTM 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact As part of the LSTM and Butantan Institute collaboration underpinned by the MRC/FAPESP and MRC Programme grant awards Dr Miyaji visits LSTM annually to disseminate results findings and discuss next steps of this collaboration.

TBC
Year(s) Of Engagement Activity 2013,2014,2015
 
Description Eliane Miyaji - visit and talk at LSTM 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach Regional
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact Eliane Miyaji (Butantan Institute) visited the Liverpool School of Tropical Medicine (LSTM) on the 4rd and 5th of June of 2013 to meet Stephen Gordon and Daniela Ferreira and to discuss results related to the Me vaccine project (MR/K01188X/1). During this visit she has presented a talk on "Pneumococcal vaccines based on PspA and other choline-binding proteins" that was attended by 20 researchers from LSTM, University of Liverpool and John Moores University.

This visit has raised the profile of Butantan Institute and Eliane has had the opportunity to discuss further collaborations. As a result Imran Saleem (John Moores University) is preparing a grant proposal in collaboration with Eliane Miyaji and Viviane Goncalves (Butantan Institute) under the BBSRC/FAPESP bilateral agreement
Year(s) Of Engagement Activity 2013
URL http://www.lstmliverpool.ac.uk/events/june-2013/4-june-eliane-miyaji/
 
Description Facebook page 
Form Of Engagement Activity Engagement focused website, blog or social media channel
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact Facebook page advertises studies and informs on latest study developments, currently over 60 members and a footprint of over 1000 people
Year(s) Of Engagement Activity 2015
URL https://www.facebook.com/LivRRC/?ref=aymt_homepage_panel
 
Description Human Challenge Models workshop 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Lancet Infectious Diseases report as per papers

MRC Programme grant Expt Human Pneumococcal Carriage funded Nov 2014
Year(s) Of Engagement Activity 2013
 
Description Medical Research Council - UK-Brazilian Infectious Disease workshop 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Geographic Reach International
Primary Audience Postgraduate students
Results and Impact The objective of the workshop will be 2 fold, not only to identify specific individual collaborations but also to scope out and inform the joint UK-Brazilian call in the area of infectious disease, which will be launched under the Newton Fund next year.

scope out and inform the joint UK-Brazilian call in the area of infectious disease, which will be launched under the Newton Fund next year.
Year(s) Of Engagement Activity 2014
 
Description Presentation to Undergraduate and Post Graduate Students at the University of Sao Paulo 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Undergraduate students
Results and Impact Daniela Ferriera gave a talk about how controlled human infection model can accelerate Pneumococcal Vaccine development to undergradiate and postgraduate students.
Year(s) Of Engagement Activity 2017
 
Description Twitter Feed (@Liv_RRN) 
Form Of Engagement Activity Engagement focused website, blog or social media channel
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact Twitter feed used to encourage study participation and inform donors and collaborators of current policy
Year(s) Of Engagement Activity 2015
 
Description University of Tokyo - EHPC collaboration meeting 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact A meeting between LSTM and Tokyo university to discuss the a joint grant application to the MRC DPFS as part of a new collaboration to develop mucosal vaccines based on PspA antigen. This collaboration resulted from the visit of Dr Ferreira to Butantan Institute (Sao Paulo) as part of the MRC/FAPESP award.
Year(s) Of Engagement Activity 2015
 
Description XII European Meeting on the Molecular Biology of the Pneumococcus, Europneumo, UK 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Talk to disseminate research findings during this conference. Breakfast meeting with approximately 20 EHPC collaborators to discuss the EHPC Programme planned activities.
Year(s) Of Engagement Activity 2015