Optimmunize: imprOving the beneficial effects Of vaccines 19 - 21 february 2020

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Name:

Optimmunize: Improving the

beneficial effects of vaccines

Wellcome Genome Campus Conference Centre, Hinxton, Cambridge, UK

19-21 February 2020

Scientific Programme Committee:

Christine Stabell Benn

University of Southern Denmark, Denmark

Rebecca Chandler

Uppsala Monitoring Centre, Sweden

Nigel Curtis

The University of Melbourne, Australia

Eleanor Fish

University of Toronto, Canada

Sabra Klein

Johns Hopkins University, USA

Tweet about it: #optimmunize20

@ACSCevents /ACSCevents /c/WellcomeGenomeCampusCoursesandConferences

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Scientific Programme Committee

Christine Stabell Benn

University of Southern Denmark, Denmark

Rebecca Chandler

Uppsala Monitoring Centre, Sweden

Nigel Curtis

The University of Melbourne, Australia

Eleanor Fish

University of Toronto, Canada

Sabra Klein

Johns Hopkins University, USA

Wellcome Genome Campus Scientific Conferences Team:

Jemma Beard

Conference and Events

Organiser

Nicole Schatlowski

Scientific Programme

Officer

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Dear colleague,

I would like to offer you a warm welcome to the Wellcome Genome Campus Advanced Courses and

Scientific Conferences: Optimmunize: Improving the beneficial effects of vaccines. I hope you will find

the talks interesting and stimulating, and find opportunities for networking throughout the schedule.

The Wellcome Genome Campus Advanced Courses and Scientific Conferences programme is run on a

not-for-profit basis, heavily subsidised by the Wellcome Trust.

We organise around 50 events a year on the latest biomedical science for research, diagnostics and

therapeutic applications for human and animal health, with world-renowned scientists and clinicians

involved as scientific programme committees, speakers and instructors.

We offer a range of conferences and laboratory-, IT- and discussion-based courses, which enable the

dissemination of knowledge and discussion in an intimate setting. We also organise invitation-only

retreats for high-level discussion on emerging science, technologies and strategic direction for select

groups and policy makers. If you have any suggestions for events, please contact me at the email

address below.

The Wellcome Genome Campus Scientific Conferences team are here to help this meeting run

smoothly, and at least one member will be at the registration desk between sessions, so please do

come and ask us if you have any queries. We also appreciate your feedback and look forward to your

comments to continually improve the programme.

Best wishes,

Dr Rebecca Twells Head of Advanced Courses and Scientific Conferences rebecca.twells@wellcomegenomecampus.org

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General Information

Conference Badges

Please wear your name badge at all times to promote networking and to assist staff in identifying you.

Scientific Session Protocol

Photography, audio or video recording of the scientific sessions, including poster session is not

permitted.

Social Media Policy

To encourage the open communication of science, we would like to support the use of social media at

this year’s conference. Please use the conference hashtag #optimmunize20. You will be notified at

the start of a talk if a speaker does not wish their talk to be open. For posters, please check with the

presenter to obtain permission.

Internet Access

Wifi access instructions:

Join the ‘ConferenceGuest’ network

Enter your name and email address to register

Click ‘continue’ – this will provide a few minutes of wifi access and send an email to the

registered email address

Open the registration email, follow the link ‘click here’ and confirm the address is valid

Enjoy seven days’ free internet access!

Repeat these steps on up to 5 devices to link them to your registered email address

Presentations

Please provide an electronic copy of your talk to a member of the AV team who will be based in the

meeting room.

Poster Sessions

Posters will be displayed throughout the conference. Please display your poster in the Conference

Centre on arrival. There will be two poster sessions during the conference.

Odd number poster assignments will be presenting in poster session 1, which takes place on

Wednesday, 19 February, at 18:00 – 19:00.

Even number poster assignments will be presenting in poster session 2, which takes place on

Thursday, 20 February, at 18:00 – 19:00

The page number of your abstract in the abstract book indicates your assigned poster board

number. An index of poster numbers appears in the back of this book.

Conference Meals and Social Events

Lunch and dinner will be served in the Hall, apart from lunch on Wednesday, 19 February when it will

be served in the Conference Centre, alongside registration. Please refer to the conference

programme in this book as times will vary based on the daily scientific presentations. Please note there

are no lunch or dinner facilities available outside of the conference times.

All conference meals and social events are for registered delegates. Please inform the conference

organiser if you are unable to attend the conference dinner.

The Hall Bar (cash bar) will be open from 19:00 – 23:00 each day.

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Dietary Requirements

If you have advised us of any dietary requirements, you will find a coloured dot on your badge.

Please make yourself known to the catering team and they will assist you with your meal request.

If you have a gluten or nut allergy, we are unable to guarantee the non-presence of gluten or nuts in

dishes, even if they are not used as a direct ingredient. This is due to gluten and nut ingredients being

used in the kitchen.

For Wellcome Genome Campus Conference Centre Guests

Check in

If you are staying on site at the Wellcome Genome Campus Conference Centre, you may check into

your bedroom from 14:00. The Conference Centre reception is open 24 hours.

Breakfast

Your breakfast will be served in the Hall restaurant from 07:30 – 09:00.

Telephone

If you are staying on-site and would like to use the telephone in your room, you will need to

contact the Reception desk (Ext. 5000) to have your phone line activated – they will require your

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Departures

You must vacate your room by 10:00 on the day of your departure. Please ask at reception for

assistance with luggage storage in the Conference Centre.

Taxis

Please find a list of local taxi numbers on our website. The conference centre reception will also be

happy to book a taxi on your behalf.

Return Ground Transport

Complimentary departure transport from the Conference Centre has been arranged on Friday, 21

February 2020 at the following times:

13:15 to Stansted and Heathrow airports.

13:30 to Cambridge train station and city centre (Downing Street).

A sign-up sheet will be available at the conference registration desk from 15:30 on Wednesday, 19

February until Thursday, 20 February. Please note that places are limited and will be allocated on a

first come first served basis.

Please allow a 30-40 minute journey time to both Cambridge and Stansted Airport, and two and a

half hours to Heathrow.

Messages and Miscellaneous

Lockers are located outside the Conference Centre toilets and are free of charge.

All messages will be available for collection from the registration desk in the Conference Centre.

A variety of toiletry and stationery items are available for purchase at the Conference Centre

reception. Cards for our self-service laundry are also available.

Certificate of Attendance

A certificate of attendance can be provided. Please request one from the conference organiser

based at the registration desk.

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Contact numbers

Wellcome Genome Campus Conference Centre – 01223 495000 (or Ext. 5000)

Wellcome Genome Campus Conference Organiser (Jemma) – 07473 934631

If you have any queries or comments, please do not hesitate to contact a member of staff who will

be pleased to help you.

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Conference Summary

Wednesday, 19 February

11:30 – 12:50 Registration with buffet lunch

12:50 – 13:00 Welcome and introduction

13:00 – 14:30 Session1: Why the no-specific effects of vaccines are important

14:30 – 15:30 Keynote lecture: Mihai Netea, Radboud University, The Netherlands

15:30 – 16:00 Afternoon tea

16:00 – 17:30 Session 2: Potential non-specific effects of vaccines – evidence from clinical

trials

17:30 – 18:00 Lightning talks

18:00 – 19:00 Poster session 1 (odd numbers) and drinks reception

19:00 – 21:00 Dinner

Thursday, 20 February

09:00 – 10:45 Session 3: Effects of vaccines on immunity and the epigenome

10:45 – 11:15 Morning coffee

11:15 – 12:30 Session 4: Sex-differnces in immunity and response to vaccines

12:30 – 14:00 Lunch

14:00 – 15:30 Session 5: Potential off-target use of vaccines

15:30 – 16:00 Afternoon tea

16:00 – 17:30 Session 6: Conference Q&A

17:30 – 18:00 Lightning talks

18:00 – 19:00 Poster session 2 (even numbers) and drinks reception

19:00 – 21:00 Dinner

Friday, 21 February

09:00 – 10:00 Session 7: Vaccination of the elderly

10:00 – 10:30 Session 8: Non-specific effects of vaccines: A complex and controversial topic

10:30 – 10:45 Morning coffee

10:45 – 12:45 Session 9: Looking toward the future: New vaccines/testing products in the

pipeline

12:45 – 12:55 Closing remarks

12:55 – 13:15 Lunch

13:15 Coach departs to London Heathrow airport via London Stansted airport

12:45 Coach departs to Cambridge train station and city centre (Downing Street)

The Conference Centre hosts hundreds of one-day and residential meetings for biomedical sector clients each year, and offers preferential rates to organisers from this sector.

Facilities:

We would love to welcome you and your delegates to our venue at the heart of life-changing science.

• 300-seat auditorium with all the latest audiovisual capabilities• 8 distinctive meeting rooms with flexible set-up options for groups of 2-120 people• Large naturally-lit exhibition space with bar • 300-seat restaurant

• On-site accommodation: 134 modern and comfortable bedrooms• Outdoor space for team building activities and BBQs in summer• Complimentary parking for 180 cars and bike rack

Hold your own meeting at the Wellcome Genome Campus Conference Centre

To enquire or to book a show round please call the Sales team on 01223 495123 or email conference.centre@wellcomegenomecampus.org

www.wellcomegenomecampus.org/conferencecentre

We chose the conference facility for the beautiful surrounds, the professionalism of the staff, the high quality of the venue itself and the amazing wow factor.

Health Enterprise East Innovation Showcase”“

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Conference Sponsors

https://www.mdpi.com/journal/vaccines

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Optimmunize: Improving the beneficial effects of vaccines

Wellcome Genome Campus Conference Centre,

Hinxton, Cambridge

19 – 21 February 2020

Lectures to be held in the Francis Crick Auditorium

Lunch and dinner to be held in the Hall Restaurant

Poster sessions to be held in the Conference Centre

Spoken presentations - If you are an invited speaker, or your abstract has been selected for a

spoken presentation, please give an electronic version of your talk to the AV technician.

Poster presentations – If your abstract has been selected for a poster, please display this in the

Conference Centre on arrival.

Conference programme

Wednesday, 19 February

11:30-12:50 Registration with lunch

12:50-13:00 Welcome and Introductions

Programme Committee

13:00-14:30 Session 1: Why the non-specific effects of vaccines are important

Chair: Nigel Curtis

13:00 Opening note

Nigel Curtis

Melbourne University, Australia

13.10 Why the non-specific effects of vaccines are important

Peter Aaby

Bandim Health Project, Guinea-Bissau

13:30 Off target effects: the box is finally open but the contents are still

confusing

Adam Finn

University of Bristol, UK

13:50 Q&A & discussion

14:30-15:30 Keynote Lecture:

Trained immunity: a memory for innate host defense

Mihai Netea

Radboud University, The Netherlands

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15:30-16:00 Afternoon Tea

16:00-17:30 Session 2: Potential non-specific effects of vaccines – evidence from

clinical trials

Chair: Sabra Klein

16:00 The Melbourne Infant Study: BCG for Allergy and Infection Reduction

(MIS BAIR)

Nigel Curtis

Melbourne University, Australia

16:30 Repurposing of Vaccines: What is in a name?

Tobias Kollmann

Telethon Kids Institute, Australia

17:00 Immune system development varies according to age, location and

anemia in African children

Danika Hill

Babraham Institute, UK

17:15 Evidence for BCG-induced protection against heterologous infectious

disease in Ugandan neonates: an investigator-blind randomised

controlled trial.

Sarah Prentice

East and North Hertfordshire NHS Trust, UK

17:30-18:00 Lightning talks

18:00-19:00 Poster Session 1 (odd numbers) with drinks reception

19:00 Dinner

Thursday, 20 February

09:00-10:45 Session 3: Effects of vaccines on immunity and the epigenome

Chair: Mihai Netea

09:00 What have we learnt about BCG vaccination in the last 20 years?

(and can we improve its efficacy?)

Hazel Dockrell

LSHTM, UK

09.30 Non-specific effects of vaccines and nanoparticles: potential use in

cancer patients

Magda Plebanski

RMIT, Australia

10.00 BCG and DTPw vaccination induce altered programmes of “trained”

innate immunity in mice

David Lynn

South Australian Health and Medical Research Institute, Australia

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10.15 Exploring the role of RORa on trained immunity in human monocytes

Gizem Kilic

Radboud University Medical Center, The Netherlands

10.30 Systemic bacterial burden of sepsis-challenged neonatal mice

significantly associated with survival

Byron Brook

University of British Columbia, Canada

10:45-11:15 Morning Coffee

11:15-12:30 Session 4: Sex-differences in immunity and response to vaccines

Chair: Eleanor Fish

11:15 Age-associated sex differences in immunity to seasonal inactivated

influenza vaccines

Sabra Klein

Johns Hopkins University, USA

11:45 Vertically transferred immunity in neonates: Mothers, mechanisms and

mediators

Petra Arck

University Medical Center Hamburg-Eppendorf, Germany

12:15 Female-biased effects of aging on a chimeric hemagglutinin-based

universal influenza virus vaccine in mice

Santosh Dhakal

Johns Hopkins University, USA

12:30-14:00 Lunch

14:00-15:30 Session 5: Potential off-target use of vaccines

Chair: Christine Stabell Benn

14:00 BCG vaccines off target effects: so many good things with an emphasis

on type 1 diabetes

Denise Faustman

Massachusetts General Hospital, USA

14:30 Sex-differential effects of rabies vaccine on survival in young free-

roaming dogs, a novel animal model for the study of non-specific

effects of vaccines in high-mortality populations

Darryn Knobel

Ross University School of Veterinary Medicine, West Indies

15:00 BCG vaccination is associated with reduced malaria prevalence in

children under the age of five in sub-Saharan Africa

Mike Berendsen

Bandim Health Project, Guinea-Bissau

15:15 Discussion: How can we use off-target effects to optimize vaccination?

15:30-16:00 Afternoon Tea

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16:00-17:30 Session 6: Conference Q&A

Chair: Christine Stabell Benn

Young investigators(< 10 yrs post last degree (PhD, MD) will be asked to provide one

critical question related to the conference subject when registering. The organizers

will select the 2 best abstracts to be presented during this session. In addition, the

organizers will select the 5 most thought provoking questions, and they will be

discussed in a subsequent panel session that will include relevant speakers and the

young researchers who posed the critical questions.

17:30-18:00 Lightning talks

18:00-19:00 Poster Session 2 (even numbers) with drinks reception

19:00 Dinner

Friday, 21 February

09:00-10:00 Session 7: Vaccination of the elderly

Chair: Sarah Prentice

09:00 The impact of immunosenescence on immune responses after

influenza vaccination in older individuals

Inna Ovsyannikova,

Mayo Clinic, USA

09:20 Evidence for immunomodulatory effects of vaccination in aging

Australians

Katie Flanagan

Monash University, Australia

09:40 Panel discussion:

Can vaccines be used to combat immunosenescence?

10:00-10:30 Session 8: Non-specific effects of vaccines: a complex and

controversial topic

Chair: Eleanor Fish

10:00 Non-specific effects of vaccines: Complex and controversial….and a

challenge for current vaccine paradigms

Rebecca Chandler

Uppsala Monitoring Centre, Sweden

10:15 Discussion

10:30-10:45 Morning Coffee

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10:45-12:45 Session 9: Looking toward the future: New vaccines/testing products

in pipeline

Chair: Rebecca Chandler

10:45 Specific and off-target effects of novel live attenuated pertussis vaccine

Camille Locht

Institute Pasteur, France

11:15 Impact of adjuvants, sex, and the microbiome on SIV-vaccine-induced

immunity and protective efficacy in rhesus macaques

Marjorie Robert-Guroff

National Cancer Institute, USA

11:45 Precision adjuvants for next generation vaccines tailored to

vulnerable populations

Ofer Levy

Harvard University, USA

12:15 New live attenuated tuberculosis vaccine MTBVAC induces trained

immunity and confers protection against experimental lethal

pneumonia

Jorge Dominguez Andres

Radboud University Medical Center, The Netherlands

12:30 MV130 as a Trained Immunity-based Vaccine, clinical implications and

mechanism of action

Laura Conejero

Immunotek, Spain

12.45-12:55 Closing remarks

12:55 -13:15 Lunch

13:15 Coach depart to Heathrow Airport via Stansted Airport

13:30 Coach departs to Cambridge City Centre and Train Station

The following abstracts should not be cited in bibliographies. Materials contained herein should be treated as personal communication and should be cited as such

only with consent of the author.

2020 CONFERENCES

Evolutionary Systems Biology12-14 FebruaryOptimmunize: Improving the beneficialeffects of vaccines NEW19-21 FebruarySingle Cell Biology11-13 MarchGenomics of Brain Disorders 18-20 MarchGenomics of Rare Diseases25-27 MarchProteomics in Cell Biology and Disease Mechanisms30 March-1 AprilLongitudinal Studies20-22 AprilNursing, Genomics and Healthcare NEW27-29 AprilAntimicrobial Resistance - GenomeBig Data and Emerging Technologies4-6 MayCurating the Clinical Genome20-22 MayHealthy Ageing27-29 MayGenomic Epidemiology of Malaria 7-10 JuneVirus Genomics and Evolution15-17 JuneReproducibility, Replicability and Trustin Science NEW9-11 SeptemberGenome Informatics14-17 SeptemberCRISPR and beyond: perturbations at scale to understand genomes23-25 SeptemberGenomic Imprinting - from Biology to Disease NEW28-30 SeptemberExploring Human Host-Microbiome Interactions in Health and Disease21-23 October

COURSESLABORATORY COURSESGenomics and Clinical Microbiology19-24 JanuaryGenomics and Clinical Virology23–28 FebruaryGenetic Engineering of Mammalian Stem Cells15–27 MarchNext Generation Sequencing20–27 AprilLow Input Epigenomics NEW12-20 MayRNA Transcriptomics17-26 JuneSingle Cell Technologies and Analysis24-31 JulyMolecular Pathology and Diagnosis of Cancer22-27 NovemberDerivation and Culture of Human Induced Pluripotent Stem Cells (hiPSCs)14-18 December

COMPUTATIONAL COURSESMathematical Models for Infectious Disease Dynamics24 February-6 MarchFungal Pathogen Genomics11-16 MaySummer School in Bioinformatics22-26 JuneSystems Biology: From Large Datasets to Biological Insight6-10 JulyGenetic Analysis of Mendelian and Complex Disorders15-21 JulyProteomics Bioinformatics26-31 JulyGenetic Analysis of Population-based Association Studies21-25 SeptemberWorking with Protozoan Parasite Database Resources4-9 OctoberNext Generation Sequencing Bioinformatics18-24 OctoberComputational Systems Biology for Complex Human Disease NEW6-11 December

@ACSCeventswellcomegenomecampus.org/coursesandconferences

LECTURE/DISCUSSION COURSESClinical Genomics: Fundamentals ofVariant Interpretation in Clinical Practice29-31 JanuaryGenomic Practice for Genetic Counsellors3-5 FebruaryPractical Aspects of Small Molecule Drug Discovery21-26 JuneEvolutionary Biology and Ecology of Cancer29 June-3 JulyScience Policy: Improving the Uptake of Research into UK Policy19-21 AugustGenomics for Dermatology25-27 November

OVERSEAS COURSESNext Generation Sequencing Bioinformatics19-24 January (Chile) 9-14 February (Malaysia)Molecular Approaches to Clinical Microbiology in Africa 21-27 March (The Gambia)Genomics and Epidemiological Surveillance of Bacterial Pathogens19-24 April (Paraguay)Working with Pathogen Genomes10-15 May (Vietnam)Viral Genomics and Bioinformatics7-12 June (Uruguay)Antimicrobial Resistance of Bacterial Pathogens 27 September-3 October (Kenya)Malaria Experimental Genetics 8-13 November (The Gambia)Practical Aspects of Drug Discovery 29 November-4 December (Uruguay)

ONLINE COURSESBacterial Genomes - 4 coursesGenetic Counselling - 1 course

Please see our website for more details and scheduling of online courses

WGC_Courses_and_Conferences_2020-Abstract-Books(Blue-FullColour)december2019.indd 1WGC_Courses_and_Conferences_2020-Abstract-Books(Blue-FullColour)december2019.indd 1 03/12/2019 11:08:0403/12/2019 11:08:04

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Spoken Presentations

Why the non-specific effects of vaccines are important

Peter Aaby

Statens Serum Institut, Denmark

For public health, a vaccine serves to prevent a specific disease. Thus, the effect of a

vaccine on overall survival is assumed to be proportional to the number of specific infections

prevented. However, from measuring the overall effects of vaccines on morbidity and

mortality has come a growing number of contradictions of this assumption, and an increasing

realisation that vaccines also have important non-specific effects (NSEs). With this

knowledge, we can define new vaccination policies with more profound effects on survival.

While there is now amble evidence that vaccines not only have specific effects, we are still in

the process of understanding the NSEs, but some strong principles have emerged: First, so

far it has been shown that four live vaccines enhance resistance to unrelated infections

(beneficial NSEs) by reducing child mortality more than can be explained by prevention of

the vaccine-targeted infection. Second, in contrast, six live vaccines have deleterious NSEs,

increasing the risk of other infections, particularly for females. Third, combining live and non-

live vaccines have variable effects. Fourth, females and males react differently to vaccines.

Fifth, the sequence and combination of vaccines is important since the most recent vaccine

has the strongest NSEs. Sixth, vaccinating with live vaccines in the presence of existing

maternal immunity amplifies the beneficial NSEs. Seventh, boosting with live vaccines

amplifies the beneficial NSEs. Eight, as immune-stimulants vaccines may interact

beneficially or deleteriously with other interventions affecting the immune system. Systematic

searches for the effect of various vaccines and combinations will undoubtedly produce more

principles of NSEs in the future.

As illustrated by a few examples, public health will benefit if the NSEs are taken into

consideration. First, the live high-titre measles vaccine (HTMV) was effective in preventing

measles infection, but had to be withdrawn by WHO because it was associated with two-fold

increased female mortality. This negative effects was due to non-live vaccines being given

after the live measles vaccine. Second, campaigns with measles vaccine and live oral polio

vaccine (OPV) have played a major role in reducing child mortality in low-income countries in

the last decades. Hence, stopping live vaccines after eradication may increase mortality as

happened with smallpox vaccine and may soon happen with OPV. Third, several non-live

vaccines including the malaria vaccine (RTS,S) currently being tested in Africa have been

associated with increased female-to-male mortality. Co-administration of non-live vaccine

with a live vaccine may reduce the negative effect for girls.

S2

Notes

S3

Off target effects: the box is finally open but the contents are still confusing

Adam Finn

University of Bristol, UK

Vaccines are developed, tested, licensed and, usually, implemented based on evidence

showing their direct (on target) protective effects and safety in recipients. Evidence of their

indirect effects (impact on transmission of the target infection in the wider population) usually

follows universal implementation and often exceeds expectations. Recently models

predicting such effects have begun to be incorporated into cost-benefit models which drive

implementation and even targeting strategies (e.g. paediatric influenza and adolescent

meningococcal programmes). However the extent to which immunisation, especially in early

childhood, may have off target effects is much less clear. Epidemiological evidence of some

important downstream off-target effects (when prevention of the target infection in turn

prevents secondary infections that otherwise would have ensued e.g. varicella-group A

streptococcus; influenza-pneumococcal pneumonia; measles-multiple infections) are

beginning to emerge with supporting explanatory mechanisms in some cases. However the

picture with respect to pure off-target effects (where administration of the vaccine itself alters

the recipients’ susceptibility to unrelated infections) currently appears to be both complex

and hard-to-call. Confronted with the apparent paradox of modest efficacy of the candidate

sporozoite vaccine RTS,S against malaria alongside a numerically small but proportionally

(between groups) large safety signal with an excess of cases of meningitis, diverse in both

aetiology and timing, and even of cerebral malaria in the vaccine recipients, we developed

an alternative hypothesis that the non-live rabies vaccine, given to controls in the older study

group in whom the apparent central nervous system infection signal was observed, might

actually have been protective in some non-specific off target way. We found several studies

in the literature published over a 50 year period that support this theory as well as studies

suggesting that components of the rabies virus present in the inactivated vaccine may

activate multiple unrelated lymphocyte clones. We are currently investigating possible

mechanisms for such an effect in more detail. If proven to exist, any added protective value

of such non-specific effects of rabies vaccine might drive its introduction in communities in

which deaths from paediatric rabies cases occur but not in sufficient numbers to justify the

cost of universal prevention.

S4

Notes

S5

Trained immunity: a memory for innate host defense Mihai G. Netea, Department of Medicine, Radboud University Nijmegen Medical Center, Nijmegen, the Netherlands The inability of innate immunity to build an immunological memory, considered one of the main characteristics differentiating it from adaptive immunity, has been recently challenged by studies in plants, invertebrates, and mammals. Long-term reprogramming of innate immunity, that induces adaptive traits and has been termed trained immunity characterizes prototypical innate immune cells such as natural killer cells and monocytes, and provides protection against reinfection in a T/B-cell-independent manner. In contrast, trained immunity has been shown to be able to induce protection against reinfection in a lymphocyte-independent manner. Non-specific protective effects dependent on trained immunity have also been shown to be induced after BCG vaccination in humans. Specific signaling mechanisms including the dectin-1/Raf1 and NOD2-mediated pathways induce trained immunity, through induction of histone modifications (methylation, acetylation) and epigenetic reprogramming of monocyte function. Complex immunological and metabolic circuits link cell stimulation to long-term epigenetic reprogramming of the function of myeloid cells and their bone marrow progenitors. The concept of trained immunity represents a paradigm change in immunity and its putative role in infection and inflammation may represent the next step in the design of future vaccines and immunotherapeutic approaches.

S6

Notes

S7

The Melbourne Infant Study: BCG for Allergy and Infection Reduction (MIS BAIR) Nigel Curtis University of Melbourne, Australia

The Melbourne Infant Study: BCG for Allergy and Infection Reduction (MIS BAIR) [misbair.org.au] is a randomised controlled trial of neonatal BCG vaccination in which over 1200 newborns in Melbourne have been recruited. The clinical outcomes, measured over the first five years of life, are respiratory infections, eczema, allergic sensitisation (measured by skin prick test) and asthma. Parent-completed online questionnaires have been used to collect extensive and detailed antenatal, perinatal and postnatal data. The trial includes an extensive biobank of blood and stool samples [lifecourse.melbournechildrens.com] for laboratory studies to investigate the immunological and molecular basis for the heterologous (non-specific) effects of BCG vaccine.

.

S8

Notes

S9

Repurposing of Vaccines: What is in a name?

Tobias R. Kollmann, Nelly Amenyogbe

Telethon Kids Institute; Perth Children's Hospital; University of Western Australia

Vaccination induces changes far beyond the intended immune memory response targeting a specific pathogen. These effects have variously been labelled non-specific, off-target or heterologous. All of these terms suggest a vague, unintended impact. While that may have been true during the original discovery of effects that go beyond those targeting a specific pathogen, there now is ample data to suggest that the host modulatory functions of vaccines can specifically be harnessed to provide targeted benefit from e.g. preventing newborn mortality to reducing several non-communicable diseases. To reflect this confidence in the science associated with this stream of vaccine evaluation, we suggest use of the term 'repurposing of vaccines' to capture the active, intentional harnessing of this vast and largely untapped field of science. In analogy to the field of repurposing drugs, this name would also help demystify current research efforts on this topic and emphasize the benefits to be gained, ranging from accelerating vaccine discovery and design to increased safety to reduced R&D expenditure, thus market value.

S10

Notes

S11

Immune system development varies according to age, location and anemia in African children

Danika L. Hill 1,2‡, Edward J. Carr1,3, Tobias Rutishauser4,5, Gemma Moncunill6, Joseph J. Campo6, Silvia Innocentin1, Maxmillian Mpina4,5,7, Augusto Nhabomba8, Anneth Tumbo4,5,7, Chenjerai Jairoce8, Henriëtte A. Moll9, Menno C. van Zelm2, Carlota Dobaño6,8*, Claudia Daubenberger4,5*‡, Michelle A. Linterman1*‡.

1. Lymphocyte Signalling & Development, Babraham Institute, Cambridge, UK. 2. Department of Immunology and Pathology, Central Clinical School, Monash University and Alfred Hospital, Melbourne, Victoria, Australia 3. Department of Medicine, University of Cambridge, Cambridge, UK 4. Swiss Tropical and Public Health Institute, Basel, Switzerland. 5. University of Basel, Switzerland. 6. ISGlobal, Barcelona Centre for International Health Research, Hospital Clínic–Universitat de Barcelona, Catalonia, Spain. 7. Ifakara Health Institute, Bagamoyo, Tanzania 8. Centro de Investigação em Saúde de Manhiça, Maputo, Mozambique. 9. Department of Pediatrics, Sophia Children’s Hospital, Erasmus MC, University Medical Center, Rotterdam, the Netherlands *Joint senior authors ‡ Corresponding authors

The high incidence of infectious disease in children from low- and middle-income countries

creates a challenge for their immune systems in first few years of life. These children have

the greatest need of the protection afforded by vaccination, however vaccines often show

reduced efficacy in these populations. An improved understanding of how age, infection,

nutrition, and genetics influence immune ontogeny and function is key to informing vaccine

design for this at-risk population. We sought to identify factors that shape immune

development in children under five years of age from Tanzania and Mozambique by detailed

immunophenotyping of longitudinal blood samples collected during the RTS,S malaria

vaccine phase III trial. In this cohort, the composition of the immune system is dynamically

altered during the first years of life, and this was further influenced by geographical location,

with some immune cell types showing an altered rate of development in Tanzanian children

compared to Dutch children enrolled in the Generation R population-based cohort study.

High titer antibody responses to the RTS,S/AS01E vaccine were associated with an

activated immune profile at the time of vaccination, including an increased frequency of

antibody secreting plasmablasts and follicular helper T cells. Anemic children had lower

frequencies of recent thymic emigrant T cells, isotype-switched memory B cells and

plasmablasts, and modulating iron bioavailability in vitro could recapitulate the B cell defects

observed in anemic children. Our findings demonstrate that the composition of the immune

system in children varies according to age, geographical location and anemia status.

S12

Notes

S13

Evidence for BCG-induced protection against heterologous infectious disease in Ugandan neonates: an investigator-blind randomised controlled trial.

Prentice S (a,b), Webb EL (c) Akello F (b) Kiwudhu F (b) Akurut H (b) Elliott AM (a,b) Dockrell HM (d) Cose S (b,d) The Delayed BCG Study Team

a: Clinical Research Department, London School of Hygiene and Tropical Medicine, Keppel Street, London WC1E 7HT b: MRC/UVRI and LSHTM Uganda Research Unit, 51-59 Nakiwogo Road, Entebbe, P.O. Box 49 c: Department of Infectious Disease Epidemiology, London School of Hygiene and Tropical Medicine, Keppel Street, London WC1E 7HT d: Department of Immunology and Infection, London School of Hygiene and Tropical Medicine, Keppel Street, London WC1E 7HT

Trials conducted in low birth-weight infants in West Africa suggest that BCG vaccination

produces heterologous protection against non-tuberculous infectious disease during the

neonatal period. Uncertainty exists as to the generalisability of these findings, and thus their

broader public health implications. We conducted an investigator-blind randomised

controlled trial in 560 healthy neonates in East Africa, comparing BCG vaccination on the

day of birth with BCG vaccination at 6-weeks of age. Illness incidence during the first 10-

weeks of life was measured by active participant follow-up, comprising free access to

physician review and investigations, regular routine physician reviews and weekly telephone

interviews.

Infants randomised to receive BCG at birth had a 29% reduction in physician-diagnosed

non-tuberculous infectious disease incidence compared to BCG unvaccinated infants, during

the 6-week period prior to BCG vaccination in the delayed group (Hazard Ratio (HR) 0.71,

95% CI (0.53 to 0.95)). In pre-specified sub-group analysis, this reduction was more

pronounced in low birth-weight infants ≤2500g (HR 0.10 (0.01 to 0.75)) compared to normal

birth-weight infants (HR 0.79 (0.59 to 1.07)) interaction p-value 0.04). After BCG vaccination

in the delayed group at 6-weeks of age, there was no significant difference in physician-

diagnosed, non-tuberculous infectious disease incidence between the groups.

We conclude that BCG vaccination produces heterologous protection against non-

tuberculous infectious disease during the neonatal period, in addition to having TB specific

effects. This corroborates findings from other groups and suggests that the beneficial non-

specific effects of BCG are applicable in disparate high-mortality settings. Prioritisation of

BCG vaccination on the first day of life may have significant public health benefits through

reductions in all-cause infectious morbidity and mortality.

S14

Notes

S15

What have we learnt about BCG vaccination in the last 20 years?

(and can we improve its efficacy?)

H.M. Dockrell and S.G. Smith

London School of Hygiene & Tropical Medicine

Recent trials of new TB vaccines have used BCG vaccination as a control arm or are carried

out in populations where BCG has been given previously. These and other cross-population

observational studies that aimed to better understand the variable efficacy of BCG

vaccination have shown that although we have immunological assays that read out the

immunogenicity of BCG vaccination, we still lack proven correlates of protection. Induction of

antigen-specific Th1 responses are part of the protective immune response but not sufficient.

Antibodies and innate lymphoid cells have also been implicated. BCG vaccination has also

been shown to induce beneficial non-specific effects. Epigenetic and metabolic changes in

innate cells give rise to trained innate immunity which could explain such effects. Genetically

modified BCG vaccines, or even repeat BCG vaccination may give enhanced protection.

Exploiting the effects of particular metabolites, which by themselves appear to induce trained

immunity, may give another means of improving BCG’s efficacy. What is clear is that this old

vaccine still has much to teach us, and what we learn can inform the design of improved

vaccines for TB as well as providing protection against other infections in young children as

well as immunotherapies for use in autoimmune disease.

S16

Notes

S17

Non-specific effects of vaccines and nanoparticles: potential use in cancer patients

Magdalena Plebanski

School of Health and Biomedical Sciences, RMIT University, Melbourne, Victoria, Australia

The immune systems reactions to particulate material has been selected over evolution

based on environmental challenges, including nano and micro-sized viruses, bacteria and

parasites. However, such reactions have to be balanced against the need to maintain

homeostasis in response to internal particulate materials such as exosomes, as well as non-

lethal external particles, such as non-toxic fine and ultra-fine particulates in the air. Even as

current vaccines elicit sophisticated non-specific effects, that re-frame immune system

process, with significant impact on overall morbidity and mortality in high burden

environments, new generation vaccines engaging synthetic nanotechnology, are also likely

to trigger non-specific effects, based substantially on the rules of interaction established over

evolution between particulates and immunity, particularly at the level of antigen presenting

cells. Over the years we have mapped some of these rules, and show that nanoparticles and

microparticles used as vaccine components can elicit large non-specific effects, capable of

accelerating the clearance of infections such as malaria and influenza, as well as preventing

the elicitation of allergic airways inflammation. These non-specific effects are in turn

modulated by particle size, shape and charge. For example, dendritic cells exquisitely

internalize nanoparticles 40-60 nm, whereas other phagocytes prefer to take up

nanoparticles 500-2000 nm. After being taken up, synthetic nanoparticles also signal inside

these cells, promoting epigenetic imprinting and altered antigen presenting cell reactivity.

These studies are supporting the development of new generation synthetic nanovaccines,

with a priori defined non-specific effects, to treat cancer. We will further finalize this

presentation with considerations on the nature of immune systems alterations in cancer

patients, specifically ovarian cancer patients, which can alter their response to vaccines,

including those based on new generation synthetic nanoparticles.

S18

Notes

S19

BCG and DTPw vaccination induce altered programmes of “trained” innate immunity in mice.

David J. Lynn, Marjolein van Wolfswinkel 1, Natalie Stevens 1, Georgina Eden 1, Miriam A. Lynn 1, Damon Tumes 2

1 Computational and Systems Biology Program, Precision Medicine Theme, South Australian Health and Medical Research Institute, Adelaide, South Australia. 2 Allergy and Cancer Immunology Laboratory, Centre for Cancer Biology, University of South Australia and SA Pathology, Adelaide, South Australia. 3 College of Medicine and Public Health, Flinders University, Bedford Park, South Australia.

Vaccines prevent >2 million deaths annually by eliciting antigen-specific adaptive immunity,

however increasing evidence suggests that vaccines also elicit nonspecific effects on

morbidity and mortality unrelated to the targeted disease. Live vaccinations (e.g. BCG) are

generally associated with beneficial nonspecific effects, while some inactivated vaccines

(e.g. DTPw) have controversially been associated with increased all-cause mortality. We

hypothesised that BCG and DTPw induce altered programmes of 'trained innate immunity'

i.e. epigenetic reprogramming which alters innate responses to subsequent stimuli. To

investigate this, we immunized 4-week-old mice with BCG or DTPw alone, or with both

vaccines in two different schedules (BCG followed two weeks later by DTPw or DTPw

followed by BCG). At 12 weeks post-immunisation, we assessed cytokine responses in

monocytes and DCs by ICS, following stimulation with bacterial, fungal or viral antigens.

BCG immunisation led to significantly increased TNF production by monocytes following

stimulation with heat-killed Candida albicans (HKCA), as has been previously reported. In

contrast, DTPw immunisation induced significantly increased IL6 production by DCs

stimulated with either HKCA or LPS. Interestingly, immunisation with BCG prior to DTPw

supressed IL6 production compared to DTPw alone or mock vaccination.

Our findings indicate that immunisation of mice with BCG or DTPw induces altered

programmes of trained innate immunity altering cytokine responses to subsequent microbial

encounters. Furthermore, our data demonstrate that previously administered vaccines may

modify the programme of trained innate immunity induced by other subsequently

administered vaccines. Our ongoing studies are now investigating the transcriptomic and

epigenetic basis for the observed phenotypes.

S20

Notes

S21

Exploring the role of RORa on trained immunity in human monocytes

Gizem Kilic, Simone JCFM Moorlag, Rob ter Horst, Valerie ACM Koeken, L Charlotte J de Bree, Vera P Mourits, Leo AB Joosten, Jorge Domínguez-Andrés, Mihai G Netea

Department of Internal Medicine and Radboud Center for Infectious Diseases, Radboud University Medical Center, Nijmegen, Netherlands.

Bacille Calmette-Guérin (BCG) is a live attenuated vaccine that has been traditionally used

against tuberculosis. Besides the development of strong and fast memory B and T cell

responses upon encounter with Mycobacterium tuberculosis, recent studies have shown that

BCG vaccination is also able to improve protection against non-specific infections by

mediating epigenetic and metabolic reprogramming of innate immune cells such as

monocytes, neutrophils and NK cells, what has been termed "trained immunity". However,

the strength and degree of BCG-induced trained immunity highly varies among people and

depends on age, sex and genetic variants. Among them, the transcription factor RORα is

known for its role as a regulator in the inflammatory response in monocytes and cholesterol

metabolism. Preliminary analyses in two cohorts of healthy volunteers showed that several

SNPs related to RORα might alter inflammatory responses and immune cell training after

BCG vaccination. With this in mind, we investigated the potential role of RORα on BCG-

induced trained immunity.

To test the role of RORα on training, human monocytes isolated from buffy coat were

stimulated with BCG (5 µg/ml) in the presence and absence of RORα inhibitor SR3335 for

24 hours, then cells were washed, and fresh media were added. Following 5 days of resting

period, cells were re-stimulated with LPS for 24 hours, and cytokine levels were measured

by ELISA. To study if IL-1 signaling has role on training promoted by RORα inhibition, IL-1

signaling was blocked by interleukin 1 receptor antagonist (IL-1RA), and monocytes were

trained with BCG in presence of SR3335.

Data revealed that inhibition of RORα by SR3335 significantly increased TNFα and IL-6

cytokine production in BCG-trained monocytes, in a dose dependent manner, upon re-

challenge with LPS. Furthermore, even in the absence of training stimuli, pre-exposure of

monocytes to SR3335 elevated pro-inflammatory cytokine production following re-stimulation

of monocytes with LPS. Of note, RORα inhibition with or without TLR ligands and BCG

stimulation for 24 hours did not lead to further increase in pro-inflammatory cytokine

production, indicating that RORα inhibition does not cause acute inflammation. Lastly,

impaired IL-1 signaling did not abolish increased production in TNFα and IL-6 mediated by

RORα inhibition. Collectively, RORα was identified as a modulator of BCG-induced trained

immunity in monocytes. To understand its role and regulatory function in detail, future

studies will be focusing on whether RORα inhibition regulates monocyte metabolism and

epigenome, similar to what is observed after BCG-induced trained immunity.

S22

Notes

S23

Systemic bacterial burden of sepsis-challenged neonatal mice significantly associated with survival.

Byron Brook*, Danny Harbeson*, Nelly Amenyogbe, Rym Ben-Othman, Tobias R. Kollmann‡, Radhouane Aniba‡. *,‡ authors contributed equally

University of British Columbia, Department of Experimental Medicine.

Neonates suffer a disproportionately high rate of mortality due to infectious disease

compared to adults, and neonatal interventions have had modest success in reducing

mortality. This has largely been attributed to different neonatal specific immune responses,

but the mechanism of how neonatal systems differ from adults has not been fully explored.

Understanding what controls life and death can identify biologically important targets for

future preventative treatments like vaccines, or future therapies, and death from bacterial

sepsis can be driven by hyper-inflammation, or bacterial-induced organ damage. A neonatal

mouse model of bacterial sepsis was employed to determine what controls survival.

Observing neonatal mice was essential for age-dependent responses, but were limiting due

to mice being too small for serial sampling. To obtain systemic samples mice were

euthanized, but then final outcome could not be directly observed, therefore we required a

method of predicting outcome so that systemic responses could be correlated with survival.

Measures of health like righting reflex, degree of mobility, and weight were observed

throughout the course of disease, and final outcome noted to determine whether the health

features could be used to separate survivors from non-survivors. Each health feature was

not accurate enough on its own to predict mortality, but their combination through algorithm

and machine learning techniques led to the development of a predictor with repeated 85%

accuracy and 93% sensitivity. This predictor of mortality allowed for the euthanasia of mice,

collection of samples, and determination of whether systemic bacterial burden was

correlated with survival. Mice predicted to survive had 10 - 1000 times reduced bacterial

burden compared to non-survivors. This implicated bacterial clearance as an essential

functional mediator of outcome, and that a 1 - 3 log reduction in systemic bacterial burden

was physiologically relevant to survival. These results however do not rule out inflammation

induced damage, which is an area of study for future experiments. It is important to measure

the pathways that actually associate with life and death during the implementation of new

neonatal vaccines and therapies to increase the chance of success for the application of

new interventions to human cohorts in preventing death and saving lives. The ability to

predict mortality can be used to further explore how neonates respond to sepsis, but could

also be adapted and employed to other challenge or vaccination models.

S24

Notes

S25

Age-associated sex differences in immunity to seasonal inactivated influenza

vaccines

Sabra L. Klein,

Johns Hopkins Bloomberg School of Public Health

Influenza is an ongoing threat to human health, despite the recommended annual

vaccination. Among adults, females typically develop greater vaccine-induced immunity and

protection than males, which is caused by sex differential epigenetic modifications in X-

linked genes in B cells as well as sex steroid signaling. Furthermore, inactivated as well as

universal influenza vaccine-induced immunity declines with age, but to a greater degree in

females than males, which is associated with hormonal changes occurring during

reproductive senescence. Taken together, both sex steroids and X-linked genes affect

antibody production and results in sex-specific differences in the efficacy of vaccination

against influenza.

This work was supported by the NIH/NIAID Center of Excellence in Influenza Research and

Surveillance contract HHS N272201400007C and the NIH/ORWH/NIA Specialized Center of

Research Excellence in Sex Differences U54AG062333.

S26

Notes

S27

Vertically transferred immunity in neonates: Mothers, mechanisms and mediators

Petra Clara Arck1

1Laboratory for Experimental Feto- Maternal Medicine, Department of Gynecology and

Obstetrics, University Clinic of Hamburg- Eppendorf, Germany (p.arck@uke.de)

Early life immunity is fragile and neonates are prone to infections, which creates a major

threat to offspring’s survival and future health. A sex-specific bias has often been reported,

with boys being more susceptible to early life infections. Besides congentinal infections,

vaccine-preventable infectious diseases pose a threat to neonatal immunity, as their immune

system is still immature. However, the vulnerability during infancy can be mitigated by the

transplacental transfer of pathogen-specific antibodies and possibly other mediators of

immunity from mother to the fetus during pregnancy, followed postnatally by breast milk-

mediated immunity. Since this largely antibody-mediated passive immunity can prevent the

newborn from infections, neonatal immunity depends strongly on the maternal immune

status, including the concentration of respective specific antibodies during pregnancy.

Moreover, emerging concepts propose that mothers may transfer immunity to the newborns

via vertical transfer of immune cells. Here, a preferential differentiation of hematopoietic

stem and progenitor cells towards myelopoiesis induced by maternal immune cells could be

detected in fetal mice, along with an improved resilience against cytomegalovirus infection in

neonates. In humans, low number of maternal immune cells in cord blood have been

associated with an increased number of infections in the first year of life, primarily in male

newborns. Hence, neonatal immunity is crucially determined by the mother and adjustments

of e.g. prenatal vaccination regimes may mitigate the risk for early life infections.

S28

Notes

S29

Female-biased effects of aging on a chimeric hemagglutinin-based universal influenza virus vaccine in mice

Santosh Dhakal (1), Sharvari DeshPande (1), Meagan McMahon (2), Florian Krammer (2), Sabra L. Klein (1)

(1) Department of Molecular Microbiology and Immunology, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, USA, 21205. (2) Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY, USA, 10029.

Adult females, both in humans and mice, develop greater antibody responses to inactivated

influenza vaccines (IIV) than males, which has implications for vaccine efficacy. The female-

biased immunity to IIV diminishes with advanced age. Inactivated influenza vaccines are 10-

60% effective and, therefore, efforts to develop more efficacious influenza vaccines are

underway. Targeting the conserved stalk region of the hemagglutinin (HA) protein of

influenza viruses has shown promise as a universal influenza vaccine candidate. Whether

biological sex and age affect immunity and protection provided by universal influenza

vaccines has not been evaluated and formed the basis of this study. We used chimeric HA

(cHA)-based universal influenza vaccine constructs expressing the same H1 stalk with an

irrelevant head from H9, H11, or H12 influenza A viruses. Adult (8-10 weeks) or aged (68-70

weeks) C57BL/6 mice were immunized with influenza B virus expressing cH9/1, followed by

cH11/1 and cH12/1 protein and subsequent challenge with mouse-adapted 2009 H1N1.

Following vaccination, adult mice immunized with the chimeric HA-based universal influenza

vaccine developed significantly greater serum IgG, and IgG2c but not IgG1 antibody titers

against different group 1 influenza A viruses than vaccinated aged mice. The age-specific

decline in vaccine-induced antibody titers was significantly more pronounced in female than

male mice. After virus challenge, vaccinated aged females experiences greater morbidity

than vaccinated adult females, with age-associated changes in protection being significantly

less pronounced among males. The extent to which age-associated changes in sex steroid

hormones, including estradiol, contribute to immunosenescence is currently under

investigation. These data suggest female-biased effects of aging on cHA-based universal

influenza vaccine-induced antibody responses and protection and highlight the importance of

inclusion of sex and age in the design and dosage of diverse influenza vaccine platforms.

S30

Notes

S31

BCG vaccines off target effects: so many good things with an emphasis on type 1

diabetes

Denise Faustman

Massachusetts General Hospital, USA

Abstract unavailable at time of printing

S32

Notes

S33

Sex-differential effects of rabies vaccine on survival in young free-roaming dogs, a

novel animal model for the study of non-specific effects of vaccines in high-mortality

populations

Darryn L. Knobel1,2 (presenter), Sintayehu M. Arega1, Anne Conan1

1 Center for Conservation Medicine and Ecosystem Health, Ross University School of

Veterinary Medicine, St. Kitts (dknobel@rossvet.edu.kn)

2 Department of Veterinary Tropical Diseases, University of Pretoria, South Africa

The preponderance of epidemiological evidence in support of non-specific effects (NSEs) of

vaccines is derived from populations with high burdens of mortality due to infectious and

parasitic diseases. There are considerable methodological and ethical challenges to the

study of vaccine NSEs in these populations. We propose a novel animal model system for

the study of vaccine NSEs: populations of owned, free-roaming domestic dogs. Free-

roaming dogs are a useful model for the epidemiological study of NSEs, due to their

abundance, accessibility, high mortality rates due to infectious and parasitic diseases, and

low rates of routine vaccination. The ethical acceptability of (indeed, encouragement of)

surgical sterilization (gonadectomy) in these populations also provides avenues to

investigate the role of gonadal sex hormones in modification of vaccine NSEs by sex.

In a recent randomized controlled trial comparing the effect of animal rabies vaccine (ARV,

an inactivated, aluminum-adjuvanted vaccine) on all-cause mortality against a placebo

injection in such a population of young dogs, we found that ARV substantially increased the

hazard rate of death in female but not male dogs. We also observed that, in the placebo

group, male mortality was substantially greater than female mortality, an effect which we

presume is due to greater male susceptibility to prevalent infectious and parasitic diseases in

the population. We contend that these two findings–relatively low female compared to male

mortality in the placebo group and increased female but not male mortality in the vaccinated

group compared to the placebo group–are related and point to a plausible mechanistic

explanation. We hypothesize that the detrimental effect of ARV (and possibly other vaccines)

on female survival is mediated through an effect on the immune system that alters

susceptibility to concurrent or subsequent unrelated infections, to which females are

normally less susceptible than males due to the influence of sex-related factors (hormonal

and/or genetic). We present a causal structure of this hypothesis, and propose to test it in

populations of free-roaming dogs through factorial randomized controlled trials, stratified by

sex, that could feasibly investigate the joint effects of vaccination, treatment against

concurrent or subsequent unrelated infections, and gonadectomy on mortality rates.

Integrating measures of immunological changes following interventions will allow evaluation

of the mediating effects of the immune system. We propose that establishment of free-

roaming dogs in high-mortality populations as a novel animal model system will open up new

avenues for causal inference in the study of NSEs of vaccines.

S34

Notes

S35

BCG vaccination is associated with reduced malaria prevalence in children under the age of five in sub-Saharan Africa

Mike Berendsen MSc1,2,3,4, Sjors van Gijzel BSc1 Jeroen Smits PhD2 Quirijn de Mast MD1 Peter Aaby DMSc3,5 Christine Benn DMSc3,4 Mihai Netea MD1,6,7 Andre van der Ven MD1

1 Department of Internal Medicine, Radboud Center for Infectious Diseases, Radboud University Medical Center, Nijmegen, The Netherlands 2 Global Data Lab, Institute for Management Research, Radboud University, Nijmegen, The Netherlands 3 Research Center for Vitamins and Vaccines (CVIVA), Bandim Health Project, Statens Serum Institut, Copenhagen, Denmark. 4 Open, Odense Patient Data Explorative Network, University of Southern Denmark, Odense University Hospital, Odense, Denmark 5 Bandim Health Project, Indepth Network, Bissau, Guinea-Bissau. 6 Department for Genomics & Immunoregulation, Life and Medical Sciences Institute (LIMES), University of Bonn, Germany 7 Human Genomics Laboratory, Craiova University of Medicine and Pharmacy, Craiova, Romania

Introduction

Malaria continues to be a major cause of morbidity and mortality in sub-Saharan Africa

(SSA) without effective interventions. Bacillus Calmette-Guérin (BCG) vaccine possesses

protective non-specific effects (NSEs). This study explores whether BCG is associated with

protection against malaria in children under the age of five in SSA.

Methods

We used data from the Demographic Health Survey (DHS) program, including 34,206

children from 13 SSA countries. BCG status was taken from vaccination cards when

present; if not, mother's recall was used. Presence of malaria was defined as a positive rapid

diagnostic test (RDT). Maternally reported presence or absence of fever in the previous two

weeks defined symptomatic status. Multilevel logistic regression was used to account for the

two-stage cluster sampling method.

Results

Of the 34,206 children, 12,325 (36.0%) children were malaria positive and 29,766 (87.0%)

were BCG vaccinated. BCG vaccination was associated with a lower malaria prevalence

(adjusted Odds Ratio (aOR)=0.94, [0.90 to 0.98], especially among children of whom BCG

information was retrieved from a vaccination card (aORcard=0.88, [0.82 to 0.94]). Restricting

the analysis to children from regions with suboptimal BCG coverage increased the

association (aORcard=0.81, [0.73 to 0.89]). BCG associations were similar for asymptomatic

(aORcard=0.86, [0.81 to 0.92]) and symptomatic malaria (aORcard=0.89, [0.78 to 1.01]).

Conclusion

BCG vaccination is associated with protection against malaria. These results indicate a

possible role for timely BCG vaccination in the protection and elimination of malaria. If

confirmed in further research, our findings have substantial implications for global efforts to

reduce malaria burden.

S36

Notes

S37

The Impact of Immunosenescnece on Immune Responses after Influenza Vaccination in Older Individuals Inna G. Ovsyannikova, Richard B. Kennedy, Iana H. Haralambieva, Diane E. Grill, Gregory A. Poland

Mayo Clinic Vaccine Research Group, Mayo Clinic, Rochester, Minnesota, USA Influenza is a major threat to human health worldwide and is responsible for up to 650,000 deaths globally each year. Age and immunosenescence, and their impact on immune responses after influenza vaccination, are of increasing importance for the development of better vaccines for the elderly. The aim of this study was to examine associations between age and immunosenescence markers and vaccine-induced immune responses and identify immunosenescence-associated differences in gene expression and gene regulation in an older study population receiving seasonal influenza A/H1N1 vaccination. A cohort of 159 healthy recipients who received seasonal TIV (Fluarix) was used for this study. Baseline median HAI/VNA antibody titers (1/80; IQR 1/40-1/320) demonstrated the presence of pre-existing antibodies. A positive correlation (r=0.49) was observed between the %CD4+CD28- and the %CD8+CD28- cells and a negative correlation (r=-0.33) was found between TREC and the %CD8+CD28- cells. Telomerase (TERT) activity was positively correlated with the observed increase in the influenza-specific memory B-cell Elispot response at Day 28 compared to baseline (p=0.025). T cell receptor excision circle (TREC) levels were positively correlated with the baseline and early (Day 3) influenza A/H1N1-specific memory B-cell Elispot response (p=0.04 and 0.035, respectively). We identified an interconnected set of gene expression pathways (antigen processing/presentation, MAPK, mTOR, TCR, BCR, calcium signaling, and other cellular activities) that were associated with immunosenescence markers. Using miRNA sequencing, several miRNAs (miR-320a, miR-320b, and miR-320d) associated with markers of immunosenescence, such as age, %CD4+CD28- cells and CD4+/CD8+ T cell ratio (p<0.003), were discovered. We found several CpG methylation sites (examples include KLF14 regulating TGFβ signaling and TSPAN33 having a role in cell differentiation) associated with age within the genes controlling immune function. In conclusion, our findings provide further evidence that immunosenescence is a complex phenomenon involving multiple aspects of immunity. These data suggest that influenza-specific immunity is significantly influenced by age, and that specific markers of immunosenescence are correlated with different humoral immune response outcomes observed after vaccination in older individuals.

S38

Notes

S39

Evidence for immunomodulatory effects of vaccination in aging Australians

Katie Flanagan

Monash University, Australia

Introduction: This talk will describe preliminary results from the VITAL Trial: Vaccine

Immunomodulation Throughout the Aging Lifespan. The trial is taking a systems vaccinology

approach to study how the diphtheria-tetanus-acellular pertussis (dTap) and seasonal

influenza vaccines interact in aging Australians. It is further exploring the processes of

immunosenescence and inflammaging that occur with advancing age, and how they impact

vaccine immunogenicity. The rationale is that the elderly respond poorly to these vaccines

and that non-targeted effects of vaccines have not been explored in the elderly.

Methods: 300 adults (20-50 years) and 300 elderly (>65 years) individuals are being

randomized into one of three vaccine groups. Group 1: dTap vaccination followed by

seasonal influenza vaccination 4 weeks later; Group 2: influenza vaccination alone; Group3:

dTap plus influenza vaccination. Blood is taken at multiple timepoints for transcriptome

analysis, epigenetics, multicolour flow cytometry, cytokine multiplex and vaccine antibodies,

FBC, CRP; and stool for microbiome sequencing at baseline. Reactogenicity / adverse event

data are collected at each study visit. The trial is registered at www.clinicaltrials.gov identifier

NCT02765126.

Results: To date 264 people have been recruited into the study. Herein, we report

reactogenicity, CRP, FBC and influenza antibody data. Preliminary flow cytometry,

microbiome and transcriptome data will also be presented. Prior or concomitant dTap

negatively affects influenza antibody levels in younger males and the elderly. CRP and FBC

parameters were altered by dTap vaccination and the dual vaccination group experienced

significantly greater reactogenicity.

Conclusion: Prior or concomitant dTap vaccination impacts responses to seasonal

influenza vaccination in this interim analysis of an ongoing trial.

Disclosure of Interest Statement:

KLF has recently served on the influenza vaccine advisory boards for Seqiris and Sanofi-

Pasteur but they played no role in the design or funding for this study. She is a member of

the Australian Technical Advisory Group on Immunisation but this abstract represents her

personal views and not necessarily those of ATAGI.

S40

Notes

S41

Specific and off-target effects of novel live attenuated pertussis vaccine

Camille Locht

Center for Infection and Immunity of Lille, Inserm U1019, CNRS UMR8204, Institut Pasteur

de Lille

Pertussis, mainly caused by Bordetella pertussis, is a severe respiratory disease affecting all

age groups, but is most severe in less than 6-old months infants. While the introduction of

vaccines in the mid-20th century has led to a substantial decrease in the incidence of the

disease, it has not been controlled in any country, despite a > 85% global vaccine coverage.

In recent years many countries have even experienced a dramatic resurgence of pertussis.

Although several factors may contribute to this resurgence, the most parsimonious

explanation appears to be asymptomatic transmission of B. pertussis. None of the currently

available vaccines prevent infection by and transmission of B. pertussis, while they provide

good protection against disease. Unlike current vaccines, infection by B. pertussis induces

sterilizing immunity. We have thus developed a live attenuated pertussis vaccine to be

delivered nasally. This vaccine, named BPZE1, was shown to be safe in pre-clinical models,

including severely immunocompromised mice, and to provide strong protection against nasal

infection in both mice and non-human primates. Clinical studies have shown that it is safe in

humans, up to a dose of 109 CFU, able to transiently colonize the human respiratory tract

and to induce B- and T-cell responses to B. pertussis antigens after a single nasal delivery.

This vaccine is currently undergoing a clinical phase 2 trial involving 300 volunteers. In pre-

clinical studies we found that BPZE1 protects also against other Bordetella species, such as

Bordetella parapertussis and Bordetella bronchiseptica. Interestingly, protection against B.

bronchiseptica was based on a dual mechanism: antigen-specific inhibition of colonization,

due to cross-reactive antigens, and non-specific inhibition of B. bronchiseptica-induced

inflammation. This latter off-target effect was further investigated in Bordetella-unrelated

infectious models. BPZE1 was found to abolish lung inflammation, cytokine storm and death

induced by influenza virus A, although no cross-reactive B- or T-cell epitope exist between

the influenza virus and BPZE1. Similarly, in a murine model of RSV infection, the vaccine

showed protection that lasted for at least 8 weeks and was found to depend on BPZE1-

induced IL-17. Furthermore, in a model of non-infectious inflammation, such as allergic

asthma, vaccination with BPZE1 prior to sensitization or between sensitization and challenge

with house-dust mite prevented lung inflammation and restored the respiratory capacity of

the mice. These observations are in line with important off-target effects of live attenuated

vaccines, but, in contrast to other vaccines, BPZE1 has potent anti-inflammatory properties,

without being immunosuppressive.

S42

Notes

S43

Impact of adjuvants, sex, and the microbiome on SIV-vaccine-induced immunity and protective efficacy in rhesus macaques

Thomas Musich1, Vishal Thovarai2, Slim Fourati3, David J. Venzon4, Venkatramanan Mohanram1, Iskra Tuero1, Leia K. Miller-Novak1, Sabrina Helmold Hait1, Mohammad Arif Rahman1, Ruth Hunegnaw1, Wuxing Yuan2, Colm O’hUigin2, Mark J. Cameron5, Luca Schifanella6, Monica Vaccari6, Yongjun Sui7, Celia Labranche8, David Montefiori8, Jay A. Berzofsky7, George N. Pavlakis9, Barbara K. Felber10, Genoveffa Franchini6, and Marjorie Robert-Guroff1

1Immune Biology of Retroviral Infection Section, 4Biostatistics and Data Management Section, 6Animal Models and Retroviral Vaccines Section, and 7Molecular Immunogenetics and Vaccine Research Section, Vaccine Branch, NCI, Bethesda, MD; 2Cancer and Inflammation Program, NCI, Bethesda, MD; 3Department of Pathology and 5Department of Population and Quantitative Health Sciences, Case Western Reserve University, Cleveland, OH; 8Department of Surgery, Duke University Medical Center, Durham, NC; 9Human Retrovirus Section and 10Human Retrovirus Pathogenesis Section, Vaccine Branch, NCI, Frederick, MD.

We compared two vaccine strategies in the SIV rhesus macaque model that aimed to elicit mucosal as well as systemic immunity. Rhesus macaques were mucosally primed (first via intranasal and oral routes and next by the intratracheal route) with replicating Adenovirus SIV-recombinants encoding SIV Env and Gag. Subsequently they were boosted twice intramuscularly with ALVAC encoding SIV Gag, Pro, and Env plus Env protein in alum, or DNA encoding SIV Env, Gag, and rhesus IL-12 plus Env protein in alum. Both regimens induced strong viral-specific cellular and humoral immune responses in mucosal and systemic compartments. Following repeated low-dose intrarectal SIV challenges, significant delay of viral infection in vaccinated macaques compared to adjuvant controls that received empty Adenovirus vector plus alum only was not observed. However, delayed SIV acquisition in both the ALVAC/Env and DNA&Env vaccine arms and adjuvant controls was observed when compared to naïve controls. Transcriptomic analysis identified signatures associated with protective innate immunity. Vaccinated females that became infected, but not males, exhibited significantly lower acute viremia compared to same-sex controls, confirming our previously reported sex-difference in SIV vaccine outcomes. The rectal microbiome of females and males responded differently to the prime-boost regimen, especially following the recombinant adenovirus priming immunizations, and differentially associated with viremia control and systemic and mucosal humoral immunity. We conclude that the impact on protective efficacy of vaccine-induced microbiome alteration in males and females as well as trained innate immunity are factors of significant importance in vaccine outcome.

S44

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S45

Precision adjuvants for next generation vaccines tailored to vulnerable populations

Ofer Levy MD, PhD

Director, Precision Vaccines Program, Boston Children’s Hospital

Professor, Harvard Medical School

Associate Member, Broad Institute of MIT & Harvard

Vaccines are the most important biomedical intervention over the past century, yet their path

to discovery and development has been ad hoc, slow and frequently unsuccessful.

Moreover, distinct immunity of the young and elderly renders these populations

simultaneously more susceptible to infection as well as more difficult to successfully protect

via immunization. Next generation approaches to de-risk and accelerate effective vaccine

development include targeted clinical studies coupled with systems vaccinology to generate

hypotheses re vaccine immunogenicity, population-specific human in vitro modeling for

discovery of novel adjuvants and to characterize mechanism of action and benchmark to

licensed vaccines, as well as select animal models that . While live vaccines have the

potential to induce trained immunity and heterologous benefit, this approach is limited by

divergence of vaccine strains propagated under diverse conditions, as is noted with Bacille

Calmette-Guérin (BCG), as well as by the risk of inadvertent administration of a live vaccine

to those with primary immunodeficiencies. In this context, identification of scalable small

molecule adjuvants that are effective towards a target population is an attractive solution to

enhance vaccine effectiveness in the most vulnerable. Supported by the U.S. National

Institutes of Health/National Institute of Allergy & Infectious Diseases, the Precision Vaccines

Program has undertaken several rounds of small molecule screening against human

leukocytes that have identified several lead families targeting diverse pattern recognition

receptors. Remarkably, there is a distinct ontogeny to innate immune responses to adjuvants

and their combinations with distinct activity towards leukocytes that can be synergistic,

additive or antagonistic depending on the age of a study participant. The horizon for

progress towards next generation vaccines includes adjuvanted vaccines tailored to

vulnerable populations that enhance both specific and heterologous (“non-specific”)

immunity.

S46

Notes

S47

New live attenuated tuberculosis vaccine MTBVAC induces trained immunity and confers protection against experimental lethal pneumonia

Jorge Domínguez-Andrés, Raquel Tarancón, Santiago Uranga, Anaísa V. Ferreira, Laszlo A. Groh, Mirian Domenech, Fernando González-Camacho, Niels P. Riksen, Nacho Aguilo, José Yuste, Carlos Martín, Mihai G. Netea

Department of Microbiology, Faculty of Medicine, University of Zaragoza, Spain CIBERES and Research Network on Respiratory Diseases, Spanish Ministry of Health and Instituto de Salud Carlos III, Madrid, Spain Department of Internal Medicine and Radboud Center for Infectious diseases (RCI), Radboud University Nijmegen Medical Centre, Geert Grooteplein 8, 6500HB Nijmegen, the Netherlands. Instituto de Ciências Biomédicas Abel Salazar (ICBAS), Universidade do Porto, Porto, 4050-313, Portugal. Centro Nacional de Microbiología. Instituto de Salud Carlos III, Madrid, Spain Servicio de Microbiología, Hospital Miguel Servet, ISS Aragón, Zaragoza, Spain Department for Genomics & Immunoregulation, Life and Medical Sciences Institute (LIMES), University of Bonn, 53115 Bonn, Germany Human Genomics Laboratory, Craiova University of Medicine and Pharmacy, Craiova, Romania

Among the infectious diseases, tuberculosis is the leading cause of death worldwide, and represents a serious threat especially in developing countries. The protective effects of Bacillus Calmette-Guerin (BCG), the current vaccine against tuberculosis, have been related not only to specific induction of T-cell immunity, but also with the long-term epigenetic and metabolic reprogramming of the cells from the innate immune system through a process termed trained immunity. Here we show that MTBVAC, a live attenuated strain of Mycobacterium tuberculosis, that has been shown to be safe and immunogenic against tuberculosis antigens in adults and newborns, is also able to induce trained immunity through the induction of glycolysis and glutaminolysis, accumulation of histone methylation marks at the promoters of proinflammatory genes, facilitating an enhanced response after secondary challenge. Importantly, these findings in human primary myeloid cells are complemented by a strong MTBVAC-induced heterologous protection against a lethal challenge with Streptococcus pneumoniae in an experimental murine model of pneumonia. Collectively, our evidence underlines that MTBVAC is able to exert metabolic and epigenetic immunomodulatory effects through mechanisms similar to those of BCG. In line with this, vaccination with MTBVAC is likely to recapitulate the non-specific protective effects of BCG vaccination against heterologous infections, while at the same time improving the specific responses against Mtb infection. All together, these results underline the potential that MTBVAC has as an excellent candidate for mass vaccination, representing a possible improvement compared to the currently available BCG vaccine.

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S49

MV130 as a Trained Immunity-based Vaccine, clinical implications and mechanism of action

Antonio Nieto García 1, Paola Brandi 2, Laura Conejero 2,3, Ángel Mazón 1, María Nieto 1, Francisco J. Cueto 2, Sarai Martínez-Cano 2, Paula Saz-Leal 2,3, Michel Enamorado 2, Joaquín Amores-Iniesta 2, Miguel Casanovas 3, José Luis Subiza3, David Sancho2

1 Servicio de Alergia y Neumología Pediátrica. Hospital Universitario La Fe. Valencia, Spain 2 Centro Nacional de Investigaciones Cardiovasculares Carlos III (CNIC) Madrid, Spain. 3 Inmunotek S.L., Madrid, Spain

Background

Despite recurrent wheezing affects one third of children and is, therefore, a global health

problem with considerable expenditure and impact on quality of life, specific vaccines

licensed for its prevention are still lacking. Virtually all wheezing attacks (WA) in young

children are of viral etiology. In clinical practice, bacterial immunostimulants have offered a

strategy for the prevention of recurrent respiratory tract infections, including those of viral

origin. However, the underlying immune mechanisms are poorly understood.

Trained immunity-based vaccines (TIbV) have been recently described as those composed

of trained immunity inducers that are intended to be effective not only against the specific

pathogens targeted by the vaccine, but also against heterologous pathogens.

Aim

To evaluate the efficacy and safety of a sublingual bacterial preparation (MV130) based on

heat‐inactivated whole‐cell bacteria in preventing WA in children, and to investigate in vitro

and in experimental models, its underlying mechanism of protection.

Methods

A phase III randomized, double‐blind, placebo‐controlled, parallel‐group trial, including 120

children <3years old with ≥3 WA during the previous year was performed. They were

randomly allocated to receive active treatment (MV130) or placebo for six months; the

primary endpoint was the number of WA within one year after the first dose. The main

secondary endpoints were the duration of WA (days) and symptoms and medication scores.

Mechanisms of action of MV130 were addressed experimentally in mouse models of viral

and fungal infection and in vitro assays in human monocytes.

Results

A reduction of 40% in the number of WA in the active vs the placebo group (p<0.001) was

observed with an excellent safety profile. Secondary outcomes were also significantly

improved in the active group vs placebo. Both outcomes were significant not only during the

treatment (1st half year) but once it was discontinued (2nd half year).

MV130 induces trained immunity hallmarks in human monocytes (enhanced glycolytic

metabolism and cytokine production), and in vivo against C. albicans reinfection. In addition,

administration of MV130 in mice confers protection to intranasal viral infection mediated by

trained immunity.

Discussion

We provide evidence for the first time that a mucosal polybacterial preparation induces

trained immunity, confers protection against unrelated pathogens, and shows a quite long

lasting clinical efficacy against a virus‐related condition such as recurrent WA.

Although the TIbV concept is emerging, we believe that several current anti-infectious

vaccines and immunostimulants, including MV130, may already fall in this category.

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P1

Poster Presentations

Non-live rabies vaccine at six weeks of age increases all-cause mortality rate in

female but not male puppies: a randomized controlled trial

Sintayehu Arega, Anne Conan, Darryn Knobel

Center for Conservation Medicine and Ecosystem Health, Ross University School of Veterinary Medicine, St. Kitts.

Epidemiological evidence suggests that some non-live vaccines may have non-specific

detrimental effects in females but not males in high-mortality populations. We investigated

this proposition in a randomized controlled trial in a population of owned, free-roaming dogs.

The objectives of the trial were to evaluate whether injection of non-live rabies vaccine (RV)

in puppies at 6 six weeks of age affected survival (to all causes of mortality) through to 13

weeks of age, compared to injection of sterile water (SW), and to determine whether this

effect was modified by sex. The study design was a single-site, owner-blinded, randomized,

placebo-controlled trial in South Africa. At six weeks of age, puppies born to vaccinated

mothers (dams), were randomly assigned within litters in a 1:1 ratio to receive a

subcutaneous injection of either RV or SW. A total of 358 puppies were randomized (with

approximately equal male-to-female ratio in each group). The primary outcome was death to

all causes through seven weeks after injection. Effect size was estimated using a mixed-

effects Cox proportional hazards model stratified by body weight and adjusting for sex, with

a random effect for litter. Mortality rate was 2,664/1,000 dog-years in the RV group and

1,955/1,000 dog-years in the SW group (hazard ratio 1.35; 95% confidence intervals (CI)

0.83-2.18). The effect of RV on survival was modified by sex (p-value of likelihood ratio test

for interaction = 0.02): In the RV group relative to the SW group, female survival was

reduced (hazard ratio in females 3.09; 95% CI 1.24-7.69) while there was no evidence that

survival in males was affected (hazard ratio in males 0.79; 95% CI 0.41-1.53). We conclude

that non-live rabies vaccine at six weeks of age increased all-cause mortality in female

puppies, with no evidence of an effect in male puppies. Further randomized controlled trials

should be conducted in other high-mortality dog populations to confirm these findings. Free-

roaming dog populations provide opportunities to test additional hypotheses that may

otherwise be challenging in humans, including the effects of vaccination with live and non-

live vaccines.

P2

Non-specific effects of rabies vaccine on mortality in a Danish pig herd

Kristoffer Jarlov Jensen, Lola Kathe Tolstrup, Darryn Knobel, Charlotte Sonne Kristensen,

Gregers Jungersen, Christine Stabell Benn

The new malaria vaccine RTS,S was recently tested in phase-III randomized controlled trials

in malaria-endemic African sites. Despite a modest specific protection against malaria, girls

receiving the malaria vaccine had a worryingly 2-fold increased all-cause mortality compared

to the comparator vaccine recipients. The inactivated rabies vaccine was used as a

comparator vaccine in one of the study arms. The explanations for the adverse association

could be a female specific detrimental non-specific effect of the new non-live malaria

vaccine. Alternatively, it was hypothesised that the comparator rabies vaccine had beneficial

protective effects on mortality, corroborated by reduced meningitis incidences among the

rabies vaccine recipients. A few historical experimental murine studies and lately also an

observational study in free roaming dogs have indicated that rabies vaccine may non-

specifically reduce non-rabid mortality. Overall, whereas there is a growing body of evidence

of non-specific effects of human vaccines, there is a paucity of systematic investigations of

potential non-specific effects of veterinary routine vaccines. The present study was designed

to investigate if the veterinary inactivated rabies vaccine may have non-specific effects of

morbidity and mortality in young pigs.

In a Danish herd of commercial pigs free of wildtype rabies virus using a 2x2 factorial

randomized design, 575 pregnant sows (2-3 weeks before scheduled farrowing) and 5747 of

their subsequent offspring (median 6 days of age) were allocated (1:1) to rabies vaccine or

no rabies vaccine. Except for the rabies vaccine, all animals were treated as customary.

Death or antibiotics treatment were observed until departure from the observation stables

(approximately 12 weeks of age / 20 kgs).

Until 21 days of life, overall mortality was 1.6% (90 pigs died, rabies vaccine: n=50; control:

n=40), the mortality rate ratio being 1.27 (95% confidence interval: 0.85-1.90).

Prior sow rabies vaccination did not modify the effect of neonatal vaccination. Nonetheless,

for mortality as well as antibiotic treatments within 21 days of life, there was non-significant

indication of a beneficial effect of rabies vaccine in female piglets, but a negative effect in

male piglets from rabies-naïve sows, and a negative effect of rabies vaccine in female

piglets from rabies-vaccinated sows. These effects had waned at 12 weeks of age.

With a slightly higher mortality in the piglets receiving inactivated rabies vaccine, the study

did not lend support to the proposed beneficial effect of the rabies vaccine used in the

human malaria vaccine trial in African children. Although the low mortality impaired the

power of subgroup analyses, the study indicated an interacting effect modification by sex

and maternal vaccination.

P3

BCG vaccination induces innate immune training in adults over 50 years of age.

Mike Berendsen1,2,3, Pauli Bles1,2, Charlotte de Bree1, Kristoffer Jensen2,4, Clara Jensen5, Delfim Mendes6, Mihai Netea1,7,8, Christian Wejse9,10, Christine Benn3,5

1 Department of Internal Medicine, Radboud Center for Infectious Diseases, Radboud University Medical Center, Nijmegen, The Netherlands. 2 Bandim Health Project, Indepth Network, Bissau, Guinea-Bissau. 3 Open, Odense Patient data Explorative Network, University of Southern Denmark, Odense University Hospital, Odense, Denmark. 4 Experimental and Translational Immunology, Department of Health Technology, Technical University of Denmark, Kgs Lyngby, Denmark 5 Research Center for Vitamins and Vaccines (CVIVA), Bandim Health Project, Statens Serum Institut, Copenhagen, Denmark 6 Hospital Raoul Follereau, Bissau, Guinea-Bissau 7 Department for Genomics & Immunoregulation, Life and Medical Sciences Institute (LIMES), University of Bonn, Germany 8 Human Genomics Laboratory, Craiova University of Medicine and Pharmacy, Craiova, Romania 9 Department of Public Health, Aarhus University, Aarhus, Denmark 10 Department of Infectious Diseases, Skejby University Hospital, Aarhus, Denmark

Introduction Bacillus Calmette-Guérin (BCG) was developed as a vaccine against tuberculosis, but many studies have shown its ability to induce potent protection against other infectious diseases. These non-specific effects (NSEs) could be explained by an increased production of innate cytokines after BCG vaccination, the so called 'innate immune training', which has been seen in children and adolescents. However, it is not known if BCG induces similar responses in elderly. Methods Randomized placebo-controlled trial (RCT) among 40 HIV-negative, TB-negative, Bissau-Guinean adults above 50 years of age, randomized 1:1 to BCG-Japan or solvent. Levels of interleukin (IL)-1β, IL-6, IL-10, interferon (IFN)-γ and tumor necrosis factor (TNF)-α were measured from supernatants of peripheral blood mononuclear cells (PBMCs) stimulated with heat-killed M. Tuberculosis, Lipopolysaccharide (LPS)/Phytohaemagglutinin (PHA), S. Aureus, C. Albicans and S. Pneumoniae before immunization, 2 weeks and 2 months after immunization. Results Among the 40 participants, only 2 (5%) had visible signs of previous BCG vaccination, while 32 (80%) had a positive Quantiferon test indicating that a large part of our population had either latent or passed M. Tuberculosis infection. Two weeks after vaccination, cytokine responses in the BCG-vaccinated group were lower than the responses in the solvent group. This was, however, completely reversed at two months after vaccination, where cytokine responses were highest in the BCG-vaccinated group; an increase that was slightly higher when restricted to Quantiferon positive individuals. Conclusion This study indicates that BCG vaccination can induce innate immune training also in adults above 50 years and might be an interesting tool in counteracting immunosenescence in elderly. The fact that responses were higher in Quantiferon positive individuals suggests a boosting effect regarding the NSEs of BCG.

P4

Lifestyle Differences Between Western European and African Populations Contribute to Differential Immune Responses in the Elderly

Ozlem Bulut, Godfrey Temba, Valerie Koeken, Simone Moorlag, Charlotte de Bree, Vera Mourits, Martin Jaeger, Jorge Dominguez Andres, Andre van der Ven, Leo A.B. Joosten, Quirijn de Mast, Mihai Netea

Department of Internal Medicine, Radboud University Medical Center, Nijmegen, Netherlands

Impairment of adaptive immune responses with old age leads to higher susceptibility to

infections and increased mortality. Adaptive immunity defects are also believed to be the

underlying reason of poor vaccination efficiency in the elderly. Trained immunity is the

adaptation of innate host defense which leads to non-specific immunological memory in

innate immune cells through epigenetic and metabolic reprogramming. It is the proposed

mechanism for non-specific protective effects of vaccines. Since innate immunity is mostly

intact in the elderly, induction of trained immunity could be utilized to improve vaccination.

The aim of this study is to identify the factors affecting the innate response in the elderly with

the hope of identifying new targets for improving vaccine efficacy.

To this end, metabolomic profiles were measured by high-throughput mass spectrometry

from the plasmas of a Western European cohort of 324 healthy individuals between the ages

of 18-71 and a Tanzanian cohort of 323 healthy individuals between the ages of 18-62.

Metabolome data was analyzed using MetaboAnalyst v4.0. Out of 1376 metabolites, 57

were significantly more abundant and 77 were diminished in the elderly (>50 years old) in

both cohorts compared to younger individuals. Pathway analyses revealed that metabolites

from histidine metabolism and primary bile acid biosynthesis pathways were significantly

downregulated in the elderly in both cohorts. When the elderly of the two cohorts were

compared, 403 metabolites were significantly more abundant in Europeans. These include

metabolites contributing to oxidative stress, whereas the 398 metabolites more abundant in

the Tanzanians include metabolites with anti-oxidant and anti-inflammatory properties.

Although age-related defects are observed in the production of T-cell derived cytokines such

as IFNγ in both cohorts, there is also a lower production of monocyte-derived cytokines IL-6

and TNFα in Tanzanians in contrast to Europeans. These differences of the innate immune

response could partially be explained with the different metabolic repertoire. Most of the

metabolites which are highly different between two cohorts are food-derived and have not

yet been linked to immunological functions.

These results indicate that differences in lifestyle, especially diet, reflected by the

metabolomic signature might influence the aging of the immune system. Both unique and

shared metabolites found in the European and African cohorts would provide valuable

insights to the mechanism of immune aging. The next step will be investigating the effect of

the most differentially regulated metabolites on trained immunity response in vitro and at a

later stage potentially in vivo.

P5

The influence of β-1,3-1,6-glucans on rabies vaccination titers in cats

Byrne J., Butaye P.

Ross University School of Veterinary Medicine

The aim of this study was to evaluate the immune stimulation capacity of a fully formulated

diet with β-1,3-1,6 glucans in cats by assessing the rabies antibody titer after vaccination.

Therefore, we recruited thirty-five healthy rabies antibody negative cats. The cats were

broken into two groups and fed standard diet of feed in accordance to body weight. One

group had the β-glucans incorporated; the other group served as control group. After two

weeks, of digestive adjustment and immunological stimulation, the rabies vaccine

(Imrab3TF) was administered on days 14 and 35. Blood samples were taken on days 14, 35,

and 56. Titers are determined with the Rapid Florescent Foci Inhibition Test (RFFIT). Titers

will be compared with the student t-test to determine significant differences between the

titers of the two groups.

P6

EXPLORATIVE STUDY TO THE EFFECT OF BCG BOOSTER METHODS ON THE

Priya A. Debisarun 1, Gizem Kiliç 1, Charlotte de Bree 1, Jorge Domínguez-Andrés 1, Mihai Netea 1,2,3

1. Department of Internal Medicine, Radboud University Medical Center, Nijmegen, Netherlands 2. Department for Genomics & Immunoregulation, Life and Medical Sciences Institute (LIMES), University of Bonn, 53115 Bonn, Germany 3. Human Genomics Laboratory, Craiova University of Medicine and Pharmacy, Craiova, Romania.

Rationale: the live attenuated vaccine Bacillus Calmette-Guérin is able to induce non-

specific beneficial effects and protect against non-related infectious diseases via epigenetic

and metabolic rewiring of innate

immune cells, which has been termed 'trained immunity' or innate immune memory.

However, the effect of BCG on innate immune responses wanes after three months, and the

response is highly variable between individuals. Therefore, development of methods to boost

the non-specific effects is of great importance. There are indications that the beneficial, non-

specific effects of BCG and other live vaccines may be

enhanced by means of revaccination. Albeit BCG revaccination is not a general WHO policy,

there are many countries where this is standard care. This study will investigate the effect of

BCG revaccination as well as

high-dose primary BCG vaccinations on the induction and course of trained immunity in

healthy volunteers. We hypothesize that this may increase the duration and strength of its

beneficial non-specific effects, and thereby contribute to the changing perspective on the

general immunological effects of vaccines and optimize the use of already existing vaccines.

Objective: determine the effect of BCG revaccination and high dose BCG vaccination on the

induction of innate immune memory.

Study design: a randomized controlled explorative pilot study.

Study population: 50 healthy adult volunteers.

Intervention: participants will be randomized 3:3:3:1 to four different intervention groups.

Group 1 will receive standard dose BCG vaccination followed by a subsequent standard

dose BCG vaccination three months later. Intervention group 2 will receive placebo

vaccination followed by a high dose BCG vaccination three months later. Intervention group

3 will receive placebo vaccination followed by standard dose BCG vaccination three months

later. Intervention group 4 will receive two times placebo

vaccination. Throughout the study blood will be drawn from healthy volunteers at timepoints

0, 3 months and 6 months, to analyze changes in immune responses.

Main study parameters/endpoints: comparison of cytokine responses after ex-vivo PBMC

and NK-cell re-stimulation, epigenetic and RNA transcriptome profiles.

P7

Timeliness of DTaP-IPV-Hib vaccination and development of atopic dermatitis before one year of age - A register-based cohort study of the Danish birth cohorts 1997-2012

Lise Gehrt 1, 2, Andreas Rieckmann1, 7, Nicholas Kiraly3, Aksel Karl Georg Jensen1, 4, Peter Aaby1, 5, Christine Stabell Benn1, 2, Signe Sørup1, 6

1 Research Center for Vitamins and Vaccines (CVIVA), Bandim Health Project, Statens Serum Institut, Artillerivej 5, DK-2300 Copenhagen S, Denmark 2 OPEN, Open Patient data Explorative Network, Odense University Hospital/Department of Clinical Research, University of Southern Denmark, J.B. Winsløws Vej 9 A, 3. Sal, DK-5000 Odense C, Denmark 3 Gastro and Food Allergy, Murdoch Children’s Research Institute, Royal Children's Hospital, 50 Flemington Rd Parkville, Victoria 3052 Australia 4 Section of Biostatistics, University of Copenhagen, Øster Farimagsgade 5, DK-1014 Copenhagen K, Denmark 5 Bandim Health Project, Indepth Network, Apartado 861, 1004 Bissau Codex, Guinea-Bissau 6 Department of Clinical Epidemiology, Aarhus University, Olof Palmes Allé 43-45, DK-8200 Aarhus N, Denmark 7 Section of Epidemiology, Department of Public Health, University of Copenhagen, Copenhagen, Denmark

Abstract

Background Vaccines may have non-specific effects affecting the susceptibility towards

conditions not targeted by the vaccine. A study from Australia including 4433 children found

that delayed Diphtheria- Tetanus-acellular Pertussis (DTaP) containing vaccination was

associated with a reduced risk of developing atopic dermatitis (AD) and reduced use of

prescription medicines for AD before 1 year of age. We tested if delayed vaccination with

DTaP-containing vaccines reduced the risk of developing AD before 1 year of age in Danish

children. Methods We used nationwide registers to follow 883,160 children born in Denmark

from 1997 to 2012, and to identify disease specific prescriptions and hospital contacts for AD

to define onset of AD. Binary regression models adjusting for potential confounding factors

were applied to estimate adjusted relative risks (aRR) of developing AD comparing children

with delayed vaccination against diphtheria, tetanus, acellular pertussis, polio, and

Haemophilus Influenzae type b (DTaPIPV-Hib) and children without delayed DTaP-IPV-Hib

vaccination.

Results Among 143,429 children with a delayed first dose of DTaP-IPV-Hib, 4847 (3.4%)

developed AD before 1 year of age, compared with 27,628 (3.7%) among 739,731 children

not having delayed DTaP-IPV-Hib (aRR 0.94 (95% confidence interval (CI): 0.91-0.97)). The

aRR was 0.93 (95% CI: 0.90-0.98) for children with a delayed second dose, and aRR=0.91

(95% CI: 0.87-0.95) comparing children with delayed first and second dose with all timely

vaccinated children.

Conclusion The results support the hypothesis that delayed vaccination with DTaP-IPV-Hib

is associated with reduced risk of developing AD. The dose-dependent relationship further

strengthens the evidence of a causal relationship. Bias related to health care seeking

behavior may explain some of the observed effect and further evidence from other settings is

required.

P8

Innate immune priming by Influenza vaccines

Helen R. Wagstaffe1, Anna Rodrigues Galan1, Ciara Lusa1, Jason P. Mooney1,2, Asia-Sophia

Wolf1,4, Natalie Prevatt5, Ron H. Behrens5, Eleanor M. Riley1,2, Martin R. Goodier1

1 Department of Infection biology, London School of Hygiene and Tropical Medicine. 2 The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of Edinburgh. 3 Division of Infection and Immunity, University College London. 4 Department of Clinical Research, London School of Hygiene and Tropical Medicine. Seasonal influenza vaccines are potent inducers of myeloid cell derived cytokines with the

potential to promote the responses of innate and adaptive effectors cells. Natural killer cells

are innate lymphoid effector cells which are activated by cytokines, by immune complexes and

by signals transduced via activating ligands on other host cells. After vaccination adaptive

immune responses can enhance both cytokine-driven and antibody-dependent NK cell

responses.

Our recent studies provide mechanistic insights into how influenza vaccination also primes

human natural killer cells both directly, through cell-intrinsic mechanisms, and indirectly via

the priming of NK cell activating cytokine production by myeloid cells.

An initial study of 52 adult volunteers revealed that seasonal trivalent inactivated influenza

intramuscular and intradermal and live attenuated intranasal vaccines primes NK cells for

enhanced in-vitro responsiveness (CD25 expression and IFN- production) to activating

cytokines (IL-12 and IL-18) with effects being observed up to 3 months post vaccination.

Vaccine induced enhancement demonstrated several characteristics of cytokine-induced

memory-like NK cells (CIML-NK), involves enhancement of responses within less

differentiated subsets and is reflected in vaccine effects measured directly ex-vivo.

We subsequently demonstrate in a further 51 volunteers that seasonal influenza vaccination

promotes increased myeloid cell-derived cytokine production on in-vitro re-stimulation. IL-15

alone, in the absence of vaccine antigen reveals enhancement of GMCSF, IL-10, IL-1 , TNF-

and IFN- secretion from post vaccination compared to baseline samples. Parallel

enhancement of NK cell function involves the production of IL-12 from myeloid DC.

We conclude that influenza vaccination-induced training of accessory cells combines with the

generation of CIML NK cells to enhance the overall NK cell response post-vaccination.

These studies raise several important questions surrounding vaccine induced innate immune

priming, including the impact of vaccination route and the durability of these responses.

P9

Nasal challenge with a TLR7/8 agonist (R848) induces in vivo mucosal interferons and chemokines with increased responses observed in allergy and asthma

Akhilesh Jha, Ryan S Thwaites, Tanushree Tunstall, Onn Min Kon, Robin J Shattock, Trevor T Hansel, Peter J M Openshaw

National Heart and Lung Institute, Imperial College London, UK Department of Medicine, University of Cambridge, UK Department of Infectious Diseases, Imperial College London, UK

Background

Respiratory viral infections causes significant global morbidity and mortality, especially in

vulnerable populations such as those with respiratory disease. Host recognition of viruses is

mediated in particular by Toll-like receptors (TLRs) to elicit innate immune responses. The

development of respiratory mucosal vaccine adjuvants and methods to accurately assess

respiratory mucosal immune responses are lacking.

Objectives

A synthetic analogue of single-stranded RNA (ssRNA) and TLR7/8 agonist (R848) was

administered in vivo to assess nasal mucosal innate immune responses and to study the

effect of allergy and asthma.

Methods

Nasal spray with saline and R848 was administered to healthy non-allergic (n=12), allergic

rhinitis (n=12) and allergic asthma (n=11) participants. Serial non-invasive nasal mucosal

samples and blood was collected to assess immune mediators and mucosal gene

expression.

Results

R848 was well tolerated with no evidence of systemic immune activation. R848 significantly

induced nasal mucosal IFN-α2a, IFN-γ, pro-inflammatory cytokines (TNF-α, IL-2, IL-12p70)

and chemokines (CXCL10, CCL2, CCL3, CCL4 and CCL13) compared to saline.

Participants with allergic rhinitis and asthma had increased IFN-α2a, CCL3 and CCL13

relative to healthy participants, whilst those with asthma alone had increased gene

expression of interferon stimulated genes DDX58, MX1 and IFIT3.

Conclusions

These results confirm that nasal delivery of R848 is well tolerated and innate immune

function can be assessed non-invasively and with precision. It highlights it as a tool to study

innate immune responses in airway disease and other host disease states, and has potential

for use as a vaccine adjuvant.

Future work

In-vivo and ex-vivo experiments utilising R848 will be performed to assess the epigenetic

mechanisms of innate immune memory in airway epithelial and immune cells using a single-

cell transcriptomic approach.

P10

Sex differences in immune response to influenza vaccine in healthcare workers during the 2017-2018 influenza season

Helen Kuo, Rosemary Morgan, Kathryn Shaw-Saliba, Katherine Fenstermarcher, Santosh Dhakal, Richard Rothman, Andrew Pekosz, Sabra Klein

Johns Hopkins School of Public Health

Seasonal influenza epidemics affect five to fifteen percent of the world population with an

estimated 290,000 to 650,000 annual respiratory deaths due to influenza (1). The Center for

Disease Control recommends annual influenza vaccination for healthcare workers (HCWs)

because not only does their occupation increase their risk of contracting influenza, but

HCWs care for people at higher risk for developing influenza related complications (2).

Seroconversion, a 4-fold difference between pre and post vaccination, is often used as an

indirect measurement of seroprotection against influenza post vaccination. Studies have

shown immune responses, such as seroconversion, differ between male and female vaccine

recipients, but few have focused on HCWs and their influenza vaccine responses by sex. To

understand the sex differences in immune response to the influenza vaccine in the HCWs

population, 111 participants (38 male; 73 female) were recruited during the 2017-2018

influenza season as part of the Johns Hopkins Centers for Excellence in Influenza Research

and Surveillance (JH-CEIRS) annual serological surveys among Johns Hopkins Hospital

HCWs that take part in the annual hospital-wide vaccination campaigns. Participants were

recruited just prior to receiving their annual influenza vaccine, which protects against

influenza type A/Michigan H1N1, A/Hong Kong H3N2, and B/Brisbane viruses, and blood

was collected to measure baseline antibody titer. After 28 days, blood was collected to

assess post vaccination antibody titer levels. Neutralizing antibody assays were used to

measure pre and post antibody titers against A/Michigan H1N1 and A/Hong Kong H3N2.

Pre-vaccination antibody titers were higher among those who do not seroconvert compared

to those who did seroconvert for both A strain influenza viruses. Male participants were 2.86

times more likely to seroconvert for the H1N1 antibody and 1.15 times more likely to

seroconvert for the H3N2 antibody than female participants. However, female participants

had much higher pre-vaccination antibody titer compared to male participants, suggesting

female participants were maintaining their high antibody titers throughout the year. Factors,

such as sex, BMI, age and direct patient contact were explored in a regression model as

possible correlations for female participants maintaining their pre-vaccination antibody titers.

Sex was the only statistically significant (p<0.01) factor that appears to show correlation to

seroconversion. Additional studies are needed to understand sex differences in immune

response to the influenza vaccine.

P11

A Brief Review of Adversomics: Understanding Genetic Drivers for Adverse Events

after Vaccination

Inna G. Ovsyannikova, Richard B. Kennedy, Gregory A. Poland

Mayo Clinic Vaccine Research Group, Mayo Clinic, Rochester, Minnesota, USA

Despite the huge population benefits of routine immunization, the rare occurrence of vaccine severe adverse events (SAEs) inhibits vaccine acceptance and infectious disease prevention. The molecular mechanisms underlying the immune response during SAEs after vaccination in humans are not well understood, but may be predictable rather than random. Introduced in 2009, the term “adversomics” refers to the application of immunogenetics, immunogenomics, and systems biology approaches to identifying genetic and molecular signatures for vaccine SAEs at the individual and population level. The current state of the adversomics field includes limited studies exploring genetic and proteomic signatures/biomarkers of AEs following smallpox, 17D-derived yellow fever (YF), measles-mumps-rubella (MMR), influenza and hepatitis B (HBV) vaccines. As described in published literature, genetic polymorphisms in the IL1, IL4, and IL18 genes/haplotypes and inflammatory cytokine response pathways (ICAM-1, IL-10, and CSF-3) have been associated with systemic AEs after smallpox vaccination. YF vaccine-associated disease was characterized by viremia, robust T/B cell responses, and polymorphisms in the cytokine receptor CCR5 (delta32) and its ligand RANTES (403 G/A) genes. With respect to both MMR vaccine-related febrile seizures and measles neutralizing antibody responses, GWAS associations of CD46 (rs1318653) and IFI44L (rs273259) genetic variants with febrile seizures and decreased antibodies have been recently published. Furthermore, an increased incidence of narcolepsy after adjuvanted influenza pH1N1 vaccine has been associated with HLA-DQB1*06:02/DQA1*01:02 alleles, suggesting that antigen presentation by these alleles is crucial. HBV-associated major AEs have been linked to class II HLA-DRB1* (*01:01/*03:01/*04:01/*13:01/*15:01) alleles/haplotypes and class I HLA-A2 gene interactions. Understanding and preventing vaccine SAEs is critical in order to improve public trust in

vaccine safety and to develop new safe and effective vaccines that minimize or avoid SAEs.

In this presentation I will review the current knowledge about the field of adversomics, as

well as the known associations and mechanisms of vaccine SAEs.

P12

Unconventional protection to Mycobacterium tuberculosis infection conferred by molds

Marta ROMANO, Pauline PERCIER; Giresse TIMA; Fabienne JURION and Olivier DENIS

Service Immune Response, Directorate Infectious Diseases in Humans; SCIENSANO, Brussels, Belgium

Introduction:

Molds are ubiquitous filamentous fungi and more than one hundred different indoor and

outdoor mold species have been recognized as major sources of aeroallergens that can

cause respiratory forms of allergy, ranging from allergic rhinitis to asthma.

Conflicting results have been reported on the association between allergy and infection with

M. tuberculosis and/or active Tuberculosis disease.

In this study, we wanted to address if a previous allergic sensitization with the outdoor mold

Alternaria alternata could have an effect on the susceptibility to infection with M.

tuberculosis.

Material & Methods:

For that purpose, well defined murine models of sensitization with different compounds

isolated from A. alternata or heat-killed spores were used. Intranasal instillations with these

induce lung inflammation with different profiles (differences in type of cells recruited to the

airway and in the local cytokine and chemokine expression profiles induced). Mice were

treated with the mold antigens before or after pulmonary M. tuberculosis infection.

Results:

Interestingly, our results indicate that previous sensitization with an A. alternata fraction

inducing a genuine eosinophilic / pro-Th2 response has no effect on susceptibility to

infection with M. tuberculosis, while treatment with fractions of A. alternata inducing a mixed

pro-Th2/pro-Th17 response results in an improved control of M. tuberculosis infection.

This control of pulmonary growth of M. tuberculosis is observed as early as 6 days after

infection, it is maintained for several weeks without need of continuous instillation of A.

alternata and M. tuberculosis specific adaptive immune response is not influenced by the

previous treatment with A. alternata.

In addition, treatment of M. tuberculosis infected mice with heat-killed A. alternata spores

results in a control of the bacterial burden.

Discussion-Conclusion:

Even if our results warrant further mechanistic analysis, our data collectively indicate and

confirm that the pulmonary environment can significantly shape outcome of infection with M.

tuberculosis infection.

P13

Immunological mechanisms of MV140, a polybacterial vaccine to prevent urinary tract infections

Paula Saz-Leal 1, 2, Caihong Wang 2, Marianne M. Ligon 2, Carmen M. Díez-Rivero 1, Laura Conejero 1 and Indira U. Mysorekar 2,3

1 Inmunotek, SL. Alcalá de Henares, Madrid, Spain. 2 Department of Obstetrics and Gynecology, Washington University School of Medicine, St. Louis, MO, USA. 3 Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO, USA.

Background

Urinary tract infections (UTIs) are the most reported bacterial infections in women. High

rates of recurrence lead to 5-10% of total women suffering from recurrent UTIs (rUTI).

Because of the lack of effective alternatives, antibiotic prophylaxis is still the gold standard

treatment for rUTI, contributing to the massive rise of antimicrobial resistance.

MV140 is a sublingual vaccine based on a mixture of whole-cell heat-inactivated bacteria

indicated for rUTI prophylaxis. Clinical studies in more than 1,000 subjects have

demonstrated that MV140 significantly reduces UTI episodes compared to antibiotic

prophylaxis, preventing recurrence in 80-90% of individuals during the year following

vaccination. Moreover, MV140 is effective against the specific pathogens targeted by the

vaccine, but also against heterologous infections.

Mechanistically, sublingual immunization with MV140 showed to induce a systemic CD4+ T

cell response in experimental models, but the underlying mechanisms in the context of

infection were still unknown.

Aim

To evaluate the protection conferred by the polybacterial preparation MV140 in an

experimental model of uropathogenic Escherichia coli (UPEC)-mediated UTI and to

investigate its mechanism of action. Additionally, to compare MV140 effect to that provided

by a monobacterial vaccine consisting of the specific UPEC infectious challenge.

Methods

Mice were vaccinated intranasally either with MV140 (E. coli, K. pneumoniae, E. faecalis, P.

vulgaris) or a specific UPEC strain (UTI89)-based vaccine, both heat-inactivated, and

subsequently subjected to a UTI89-induced acute UTI. Bacterial load, as well as the immune

response in urine and bladder, were measured following infection. Otherwise, the systemic

immune response to both MV140 and UPEC was evaluated in the spleens of vaccinated

versus control uninfected mice.

Results

Both MV140 and the UPEC-based vaccine conferred significant protection to infection with

an early reduction in bacterial load compared to controls. These results were accompanied

with increased numbers of myeloid immune cells in the urine, and a heightened CD4+ T cell

response in the bladder.

Both types of vaccination induced similar systemic T helper (Th) 1, Th17 and IL-10-mediated

responses against the MV140 and UTI89.

Discussion

We demonstrate for the first time that MV140 confers protection in an acute UTI

experimental model. This is accompanied by the induction of local myeloid and adaptative T

cell responses, the latter correlating with that observed systemically.

The protection conferred by MV140 against a UTI89 challenge is comparable to that

provided by its specific vaccine, and may further support a quite broad-spectrum anti-

infectious profile of this polybacterial preparation.

P14

BCG skin reactions by 2 months of age are associated with better survival in infancy: A prospective observational study

Frederik Schaltz-Buchholzer1,2,3, Mike Berendsen1,2,3,4, Adam Roth5,6, Kristoffer Jarlov Jensen1,7, Morten Bjerregaard-Andersen2,3, Marcus Kjær Sørensen2,3, Ivan Monteiro2, Peter Aaby2, Christine Stabell Benn1,2,3,8

1 Bandim Health Project, Institute of Clinical Research, Uni. Southern Denmark and Odense University Hospital, Odense, Denmark: FSB (MD), MB (MSc), CSB (Professor, DMSc). 2 Bandim Health Project, INDEPTH Network, Bissau, Guinea-Bissau: FSB, MB, KJJ (PhD), MBA (MD, PhD), MKS (medical student), IM (trial supervisor), PA (Professor, DMSc), CSB. 3 Research Centre for Vitamins and Vaccines, Copenhagen, Denmark: FSB, MB, MBA, MKS, CSB. 4 Department of Internal Medicine, Radboud Centre for Infectious Diseases, Radboud University Medical Centre, Nijmegen, The Netherlands: MB. 5 Public Health Agency of Sweden, Nobels väg 18, 171 82 Solna, Sweden: AR (MD, PhD). 6 Institution for Translational Medicine, Lund University, J Waldenströms g 35, CRC, hus 92, plan 11, Malmö, Sweden: AR. 7 Experimental and Translational Immunology, Department of Health Technology, Technical University of Denmark, Kgs Lyngby, Denmark: KJJ. 8 Danish Institute of Advanced Science, Uni. Southern Denmark, Odense, Denmark: CSB.

Objectives: Receiving Bacillus Calmette-Guérin (BCG)-Denmark vaccine at birth has been

associated with a ~40% reduction in all-cause neonatal mortality and among BCG-

vaccinated infants, developing a BCG scar during infancy has been associated with a similar

reduction in subsequent mortality. We evaluated determinants of early BCG reactions and

the reaction size by 2 months of age and tested whether having developed a BCG reaction

by 2 months and the reaction size were associated with subsequent all-cause mortality.

Design: Prospective observational study within five trials providing BCG-at-birth. The BCG

reaction status and reaction size was evaluated at home-visits by 2 months of age; mortality

up to 12 months was assessed at subsequent home-visits.

Setting: Bissau, Guinea-Bissau.

Participants: 6,012 infants BCG-vaccinated within 1 week of birth.

Main outcome measures: BCG reaction determinants were evaluated by binomial regression

providing Risk Ratios (RRs). Cox-models providing adjusted Mortality Rate Ratios (aMRRs)

were used to assess the association between 1) having a 2-month reaction (yes/no) and 2)

the reaction size tertiles and the subsequent 2-12-month mortality risk. A subgroup of infants

had their BCG reaction evaluated and were bled at age 4 weeks; samples underwent in vitro

analysis for specific and non-specific cytokine responses.

Results: 5,804 infants (97%) had developed a BCG reaction by 2 months of age; 208 (3%)

had no reaction. The BCG strain was the main determinant for both developing a 2-month

BCG reaction and the reaction size: the RR for large-reaction comparing BCG-Russia vs.

BCG-Denmark was 0.38 (0.30-0.47) and 0.61 (0.51-0.72) for BCG-Russia vs. BCG-Japan.

The 2-12-month mortality risk was 4.8% (10/208) for non-reactors, 2.9% (64/2,213) for small

reactors, 1.8% (30/1,710) for medium reactors and 0.8% (15/1,881) for large reactors. The

reactor/non-reactor aMRR was 0.49 (0.26-0.95) and there was a linear trend of decreasing

mortality with increasing reaction size (p for trend<0.001). BCG reactors had higher 4-week

specific and non-specific cytokine responses, responses that were highest among those with

large reactions.

Conclusion: Among BCG-vaccinated infants, having a BCG skin reaction by 2 months of age

was associated with markedly better survival, as was the reaction size. Our findings support

that BCG has substantial non-specific effects on all-cause mortality and indicate that

emphasising at-birth vaccination with immunogenic BCG strains and revaccinating non-

reactors could have major public health benefits.

P15

Re-vaccination with measles-mumps-rubella vaccine and hospitalization for infections lasting 2 days or longer in Denmark and Sweden – an interrupted time-series analysis

Signe Sørup, Hélène Englund, Ida Laake, Heta Nieminen, Adam Roth, Christine Benn

Research Center for Vitamins and Vaccines (CVIVA), Bandim Health Project, Statens Serum Institut, Copenhagen, Denmark (Signe Sørup and Christine Benn); Department of Clinical Epidemiology, Aarhus University, Aarhus, Denmark (Signe Sørup); Public Health Agency of Sweden, Nobels väg 18, 171 82 Solna, Sweden (Hélène Englund and Adam Roth); Department of Infectious Disease Epidemiology and Modelling, The Norwegian Institute of Public Health, Oslo, Norway (Ida Laake); Department of Public Health Solutions, Finnish Institute for Health and Welfare, Tampere, Finland (Heta Nieminen); Department of Translational Medicine, Lund University, Malmö, Sweden (Adam Roth); OPEN, Open Patient data Explorative Network, Odense University Hospital/Department of Clinical Research, University of Southern Denmark, Odense, Denmark (Christine Benn)

Background: In low-income countries, re-vaccination with live vaccines has been associated

with reduced morbidity and mortality not related to the targeted diseases. In a previous

cohort study of 4-year-old Danish children, re-vaccination with the live measles-mumps-

rubella (MMR) vaccine was associated with a 16% reduction in the rate of hospitalizations

for infections lasting 2 days or longer.

Aim: To examine if the introduction of a second MMR-dose (MMR-2) at 4 years of age in

Denmark (spring 2008) and at 6-9 years of age in Sweden (Autumn 2009) was associated

with a reduction in the rate of hospitalizations for infections lasting 2 days or longer on a

population level.

Methods: We performed an interrupted time series analysis using Poisson regression to

estimate the change in the level of the rate of hospitalizations for infections lasting 2 days or

longer following the introduction of MMR-2 adjusting for seasonality (dummy variables for

season: spring, summer, autumn, and winter). We included the 4-year olds in Denmark and

the 9-year olds in Sweden. We defined the pre-intervention, phase-in (excluded from the

model), and post-intervention periods to be March 2005 to February 2008, March 2008 to

February 2009, and March 2009 to February 2012 in Denmark and September 2005 to

August 2009, September 2009 to August 2012 and September 2012 to August 2015 in

Sweden. We estimated the observed rate of hospitalizations for infection lasting 2 days or

longer for each season and year based on the number of hospitalizations from nationwide

registers (infections identified by ICD-10 codes) and person-years at risk approximated from

population statistics.

Results: The crude rate of hospitalizations for infections lasting 2 days or longer among 4

year-olds in Denmark was 8.3 per 1000 person-years before and 7.9 per 1000 person-years

after the introduction of MMR-2. The corresponding rates among 9 year-olds in Sweden

were 2.7 and 2.5 per 1000 person-years, respectively. Although the rate decreased over the

entire study period, the Poisson regression predicted that the rate was 0.5 per 1000 person-

years higher after the phase-in of MMR-2 in Denmark (p value for level change=0.48) and

0.2 per 1000 person-years higher in Sweden (p value for level change=0.56) than what

would have occurred without the introduction of MMR-2.

Conclusion: On a population level, the introduction of MMR-2 was not associated with a

reduction in the rate of hospitalizations for infection lasting 2 days or longer in Denmark nor

in Sweden.

P16

Investigation of Mother to Child Transmission of HLA Antibodies

DM Savulescu1, M Groome2, SCK Malfeld1, S Madhi2, A Koen2, S Jones2 and M Suchard1,3

1. Centre for Vaccines and Immunology (CVI), National Institute for Communicable Diseases

(NICD), a division of the National Health Laboratory Service, South Africa

2. Respiratory and Meningeal Pathogens Research Unit (RMPRU), University of the

Witwatersrand, South Africa

3. Chemical Pathology, Faculty of Health Sciences, University of the Witwatersrand, South

Africa

Vaccination during pregnancy is a growing field of interest, however many aspects of

humoral immunity during pregnancy remain poorly understood. Human leukocyte antigens

(HLA) are proteins responsible for presentation of peptide fragments to T lymphocytes. HLA-

encoding genes are extremely polymorphic, and the foetus has a different HLA antigen

repertoire from the mother. Late in pregnancy, women produce and transfer high amounts of

antibodies, including antibodies agains HLA (HLA antibodies), to the fetus, where

presumably the antibodies may act as autoantibodies. Little is known about specificity or

transfer rate of HLA antibodies to the foetus.

We describe the prevalence of HLA antibodies in serum of thirty mother-infant pairs six

weeks post-partum. We used Luminex single angtigen technology that allows for highly

sensitive antibody detection. We found that 72% of babies and 80% of mothers expressed

HLA antibodies. In babies, 77% of HLA-I antibodies and 50% of HLA-II antibodies matched

those of their mothers, suggesting that the remaining antibodies were self-made. In contrast,

the proportion of maternal antibodies that matched those of their babies was only 16% for

HLA-I and 24% for HLA-II, suggesting that not all HLA antibodies produced during

pregnancy cross to the fetus.

Production of self-made antibodies by six weeks of age indicates early capacity of the

neonatal immune system to generate HLA antibodies; either natural antibodies or preceded

by exposure to specific HLA antigens. Our findings further indicate that there is selective

antibody transfer of particular antibody specificities from mother to infant. Because HLA

antibodies may act against paternal antigens present in the neonate, the mechanism of

selective antibody transfer or depletion may give insights into mechanisms of infant immune

tolerance.

P17

Sex-specific effects of early life influenza vaccination on protection against diverse influenza viruses in mice

Rebecca L Ursin, Yishak Woldetstadik, Georgia Stavrakis, Kristyn E. Sylvia, Harrison R. Powell, Hsuan Liu, Andrew Pekosz, and Sabra L. Klein

Johns Hopkins University Bloomberg School of Public Health, Departments of Biochemistry and Molecular Biology, and the W. Harry Feinstone Department of Molecular Microbiology and Immunology, Baltimore, MD 21205

Influenza viruses kills tens of thousands of people annually worldwide, with seasonal

influenza vaccines serving as the best mode for protection. Vaccine efficacy depends on a

myriad of factors, including intrinsic characteristics of the virus, such as mutations that

enable viruses to escape host immunity, as well as host factors, including sex, age, and

immune history. To test the hypothesis that early life exposure to influenza vaccines may

differentially affect responses to vaccination and infection later in life, we immunized and

boosted male and female C57BL/6 mice beginning at postnatal day (PND) 11-14 with a 2009

H1N1 inactivated influenza vaccine (IIV) followed by re-vaccination in adulthood (PND 63-

75) and subsequent infection with a 2009 H1N1 drift variant virus. Prior to puberty, both

males and females mounted low neutralizing antibody (nAb) titers, with no sex differences in

vaccine-induced immune responses. Upon vaccination in adulthood, robust, sex-

differentiated responses were observed in which females had greater nAb titers than males

and were better protected following challenge with the drift variant virus. To determine the

range of cross-neutralization and protection against mutating influenza viruses, we

developed a panel of 2009 H1N1 viruses that contain the following mutations in the

immunodominant hemagglutinin (HA) protein: a single amino acid mutant virus with a

mutation at position K166Q; a double mutant at positions K166Q and G157E; a triple mutant

with K166Q, G157E, and N211D; and a virus with the K166Q mutation plus an entire

antigenic region substituted with a non-human HA. Using these viruses, we determined that

regardless of the mutations in HA, unvaccinated adult females experience greater morbidity

and mortality and have a lower lethal dose 50 (LD50) than their male counterparts. Whether

immunization with IIV prior to puberty and/or in adulthood differentially affects cross-

neutralization and protection against the panel of 2009 H1N1 viruses that contain HA

mutations is currently under investigation. These studies are critical for understanding how

pre-existing immunity affects protection against viral infection and will help to develop better

influenza vaccines, including universal influenza vaccines, that provide equal protection

against circulating strains of influenza viruses in both males and females.

P18

Towards a Vaccine Based on Enterococcal Membrane Vesicles

Theresa Wagner (1), Mona Johannessen (1), Kristin Hegstad (1,2)

(1) Research group for Host-Microbe Interactions, Department of Medical Biology, Faculty of Health Sciences, University of Tromsø – The Arctic University of Norway (2) Norwegian National Advisory Unit on Detection of Antimicrobial Resistance, Department of Microbiology and Infection Control, University Hospital of North-Norway, Tromsø, Norway

Enterococcus faecium is a leading cause of nosocomial infections, especially in

immunocompromised patients. Multi-resistant E. faecium, including Vancomycin Resistant

Enterococci (VRE) impair therapy and limit treatment options. Thus, novel therapeutic

approaches and alternatives for infection control are desperately needed to combat VRE.

Vaccines are promising alternatives in times of increasing antimicrobial resistance, since the

risk of resistance development is low and vaccines can be used prophylactically. For

enterococci, there are no available vaccines yet, even though research has been ongoing to

identify vaccine candidates.

In a previous study, we isolated and characterized membrane vesicles (MVs) produced by E.

faecium. We described their proteomic content in four clinically relevant strains, including

VRE. The E. faecium MV contained known virulence factors, among them adhesins,

collagen binding proteins, fibronectin binding protein, as well as biofilm promoting factors.

Also, antimicrobial resistance related proteins were found to be MV-associated, including all

proteins of the vanA-cluster of a VRE. Furthermore, we found six described vaccine

candidates to be associated with enterococcal MVs.

MVs would be auspicious vaccine candidates since they are highly stable, non-infectious

and non-replicative. They contain major immunogenic proteins, and would be able to elicit

protective immune responses and act as adjuvant.

Since high amounts of pure MVs will be needed for vaccination studies, we optimized the

production of MV, by investigating the influence of antibiotic exposure, as well as different

isolation methods, on the outcome. The cytotoxicity of MVs was tested in various cell lines.

At present we are immunizing rabbits with E. faecium MV and are planning to characterize

their serum.

In the future, we would like to explore the potential use of MVs to elicit protective immunity

against E. faecium, including VRE, infection.

P19

Notes

P20

Notes

Delegate List

Peter Aaby

Statens Serum Institut

p.aaby@bandim.org

Asimenia Angelidou

Boston Children's Hospital

Asimenia.Angelidou@childrens.harvard.e

du

Petra Arck

University Medical Center Hamburg

p.arck@uke.de

Sintayehu Arega

Ross University School of Veterinary

Medicine

sintayehuarega@students.rossu.edu

Christine Stabell Benn

University of Southern Denmark

cb@ssi.dk

Mike Berendsen

Bandim Health Project

mike.berendsen@radboudumc.nl

Byron Brook

University of British Columbia

bnfb89@hotmail.com

Ozlem Bulut

Radboud University Medical Center

ozlem.bulut@radboudumc.nl

Egle Butkeviciute

LSHTM

egle.butkeviciute@lshtm.ac.uk

Jake Byrne

Ross University School of Veterinary

Medicine

jakebyrne@students.rossu.edu

Rebecca Chandler

Uppsala Monitoring Centre

rebecca.chandler@who-umc.org

LAURA CONEJERO

INMUNOTEK

lconejero@inmunotek.com

Maria Giulia Conti

Sapienza University of Rome

mariagiulia.conti@uniroma1.it

Nigel Curtis

University of Melbourne

nigel.curtis@rch.org.au

Mia Damhus

Center for Nutrition and Therapy

md@cetcenter.dk

Joseph de Vrieze

Oost Indische Inkt

jopdevrieze@gmail.com

Priya Debisarun

RadboudUMC

priya.debisarun@radboudumc.nl

Santosh Dhakal

Johns Hopkins University

sdhakal3@jhmi.edu

Hazel Dockrell

London School of Hygiene & Tropical

Medicine

hazel.dockrell@lshtm.ac.uk

Jorge Dominguez Andres

Radboudumc

jorge.dominguezandres@radboudumc.nl

David Elliman

PHE/GOSH

david.elliman@nhs.net

Jorge Esparza Gordillo

GlaxoSmithKline

jorge.x.esparza-gordillo@gsk.com

Denise Faustman

Harvard Medical School

faustman@helix.mgh.harvard.edu

Zoltan Fehervari

Nature Research

z.fehervari@nature.com

Adam Finn

University of Bristol

Adam.Finn@bristol.ac.uk

Eleanor Fish

University of Toronto

en.fish@utoronto.ca

Katie Flanagan

UTAS/RMIT/Monash University

katie.flanagan@ths.tas.gov.au

Anna Fletcher

Micropathology LTD.

a.fletcher@micropathology.com

PILAR GARCA CORBEIRA

GlaxoSmithKline

eventos.gsk@bcdme.es

Lise Gehrt

University of Southern Denmark

ligh@ssi.dk

MARTIN R GOODIER

LSHTM

martin.goodier@lshtm.ac.uk

Danika Hill

Babraham Institute

danika.hill@babraham.ac.uk

Akhilesh Jha

University of Cambridge

aj580@cam.ac.uk

Kamilla Josefsdottir

Directorate of Health

kamilla@landlaeknir.is

Konstantinos Karampatsas

St George's University of London

kostaskarab@gmail.com

Christine Karanja Chege

Kenyatta University

christinechege@gmail.com

Gizem Kilic

Radboudumc

gizem.kilic@radboudumc.nl

Sabra Klein

Johns Hopkins University

sklein2@jhu.edu

Henrik Kloverpris

Africa Health Research Institute

henrik.kloverpris@ahri.org

Darryn Knobel

RUSVM

dknobel@rossvet.edu.kn

Tobias Kollmann

Telethon Kids Institute

tkollm@mac.com

Helen Kuo

Johns Hopkins School of Public Health

hkuo8@jhu.edu

Ofer Levy

Boston Children's Hospital

ofer.levy@childrens.harvard.edu

Camille Locht

Inserm

camille.locht@pasteur-lille.fr

David Lynn

SAHMRI

david.lynn@sahmri.com

Anne Marie Rosendahl Madsen

University of Southern Denmark

arosendahl@health.sdu.dk

Siddhartha Mahanty

University of Melbourne

smahanty@unimelb.edu.au

Hafza Zahira Manzoor

University of Veterinary and Animal

Sciences

zahra.manzoor17@gmail.com

Francois Mayer

Nature Microbiology

francois.mayer@nature.com

Victoria McLelland

ION Osteopaths

victoriamclelland@yahoo.com.au

Kirsty Minton

Nature Reviews Immunology

k.minton@nature.com

Jessica Mwinyi

Swedish Medical Products Agency

jessica.mwinyi@mpa.se

Mihai Netea

Radboud UMC Nijmegen

mihai.netea@radboudumc.nl

Inna Ovsyannikova

Mayo Clinic

ovsyannikova.inna@mayo.edu

Magdalena Plebanski

RMIT University

Magdalena.plebanski@rmit.edu.au

Katrina Pollock

Imperial College London

k.pollock@imperial.ac.uk

Sarah Prentice

East and North Hertfordshire NHS Trust

sarah.prentice@nhs.net

Marjorie Robert Guroff

National Institutes of Health

guroffm@mail.nih.gov

Marta Romano

ISP-WIV Belgium

marta.romano@sciensano.be

Christine Rueckert

CureVac AG

booking@curevac.com

PAULA SAZ

INMUNOTEK

psaz@inmunotek.com

Frederik Schaltz-Buchholzer

Bandim Health Project

buchholzer@gmail.com

Klaus Schwamborn

Valneva

klaus.schwamborn@valneva.com

Frank Shann

University of Melbourne

shannf@netspace.net.au

Sven Arne Silfverdal

Umea University

sven.arne.silfverdal@umu.se

Signe Soerup

Statens Serum Institut AND Aarhus

UNiversity

sgs@ssi.dk

Adrian Spillmann

Valneva

adrian.spillmann@valneva.com

Melinda Suchard

National Institute for Communicable

Diseases

suchardmelinda@gmail.com

Rebecca Ursin

Johns Hopkins University

rursin1@jhu.edu

Theresa Wagner

University of Tromsø

theresa.wagner@uit.no

Hilton Whittle

LSHTM

hcwhittle@yahoo.co.uk

Index

Aaby, P S1 Robert-Guroff, M S43

Arck, P S27 Romano, M P12

Arega, S P1

Saz, P P13

Benn, C. S P2 Schaltz-Buchholzer, F P14

Berendsen, M S35, P3 Soerup, S P15

Brook, B S23 Suchard, M P16

Bulut, O P4

Byrne, J P5 Ursin, R P17

Conejero, L S49 Wagner, T P18

Curtis, N S7

Debisarun, P P6

Dhakal, S S29

Dockrell, H S15

Dominguez Andres, J S47

Faustman, D S31

Finn, A S3

Flanagan, K S39

Gehrt, L P7

Goddier, M P8

Hill, D S11

Jha, A P9

Kilic, G S21

Klein, S S25

Knobel, D S33

Kollmann, T S9

Kuo, H P10

Levy, O S45

Locht, C S41

Lynn, D S19

Netea, M S5

Ovsyannikova, I S37, P11

Plebanski, M S17

Prentice, S S13

A

Hinxton Hall

B

C

Hinxton HallPompeiian RoomLibrary RoomGreen RoomRestaurantLounges/BarBedrooms 362-367/407-410

Conference CentreReception

Francis Crick AuditoriumJames Watson Pavilion

Rosalind Franklin PavilionLoft Room 1 and 2

Willow Court (A&B)B = 230-243

330-343A = 244-259

344-361

Tennis CourtTraining Suite

Mulberry Court (C)201-229301-329401-406

Fire Assembly Point

Designated Smoking Area

To Hinxton Village(Vehicle access via main exit to site)

Disabled

The SulstonLaboratories

EMBL - EBI

WestPavilion

RSF

The DataCentre

Wet Labs

The

Mo

rgan

Bu

ildin

g

EBI South

Reception

The CairnsPavilion

Conference Centre

@ACSCevents Wellcome Genome Campus Courses and Conferenceswellcomegenomecampus.org /coursesandconferences