1 The SAPHIR partners met in June 2017 in Lelystad to share their new results and discuss their next plans. Promising and exciting results have been obtained so far. To name a few: • protection by a one shot subunit BRSV vaccine has been achieved in calves under maternal immunity, • new antigen candidates for a Mycoplasma bovis vaccine have been identified, • transgenic Eimeria lines inducing cross-protective immunity have been produced, • the safety profile of an attenuated PRRSV vaccine has been documented, • adaptable PRRSV attenuated viruses have been constructed, • DNA vaccines against PRRSV presenting very encouraging immunogenicity profiles have been developed. The ex vivo cell response to molecularly-defined adjuvants revealed mononuclear phagocyte subset-dependent and age-dependent patterns, that will lead to tailored in vivo evaluations in the next period, especially in the context of Mycoplasma hyopneumoniae and PRRSV vaccines. A very innovative approach of the modeling of the immune response has been designed and validated in the case of PRRSV infection in pigs. A large genetic and genomic analysis of samples from vaccinated pigs and chicken is ongoing for the identification of biomarkers of good vaccine response. Importantly, sociologic drivers of vaccine use have been identified from European and Chinese interviews and the economic impact of several SAPHIR diseases has been estimated at farm and national levels from the literature and data bases, providing bases for building business cases of several SAPHIR vaccines. Additional results are to come, including efficacy assessments in close to the field conditions and safety evaluations for bringing some of the SAPHIR vaccines to the market. Isabelle Schwartz‐Cornila Project Coordinator STRENGTHENING ANIMAL PRODUCTION AND HEALTH THROUGH THE IMMUNE RESPONSE ISSUE 4 November 2017 About SAPHIR Strengthening Animal Production and Health through the Immune Response” (SAPHIR) is a €10.7 million project funded by the EU and the Swiss Government. SAPHIR brings together interdisciplinary expertise from 14 academic institutes including a Chinese partner, 5 SMEs, 1 pharmaceutical company and EFFAB. SAPHIR is a H2020 project running from 1 March 2015 to 28 February 2019. SAPHIR 1 NEWS 2 PROFILES 12 EVENTS 13 PUBLICATIONS 14 CONTACT 14 SAPHIR: T hrough Innovative Approach! NEWSLETTER
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The SAPHIR partners met in June 2017 in Lelystad to share their new results and discuss their next plans. Promising and exciting results have been obtained so far. To name a few:
• protection by a one shot subunit BRSV vaccine has been achieved in calves under maternal immunity,
• new antigen candidates for a Mycoplasma bovis vaccine have been identified, • transgenic Eimeria lines inducing cross-protective immunity have been
produced, • the safety profile of an attenuated PRRSV vaccine has been documented, • adaptable PRRSV attenuated viruses have been constructed, • DNA vaccines against PRRSV presenting very encouraging immunogenicity
profiles have been developed.
The ex vivo cell response to molecularly-defined adjuvants revealed mononuclear phagocyte subset-dependent and age-dependent patterns, that will lead to tailored in vivo evaluations in the next period, especially in the context of Mycoplasma hyopneumoniae and PRRSV vaccines.
A very innovative approach of the modeling of the immune response has been designed and validated in the case of PRRSV infection in pigs. A large genetic and genomic analysis of samples from vaccinated pigs and chicken is ongoing for the identification of biomarkers of good vaccine response.
Importantly, sociologic drivers of vaccine use have been identified from European and Chinese interviews and the economic impact of several SAPHIR diseases has been estimated at farm and national levels from the literature and data bases, providing bases for building business cases of several SAPHIR vaccines. Additional results are to come, including efficacy assessments in close to the field conditions and safety evaluations for bringing some of the SAPHIR vaccines to the market.
Isabelle Schwartz‐Cornila
Project Coordinator
STRENGTHENING ANIMAL PRODUCTION AND HEALTH THROUGH THE IMMUNE RESPONSE
ISSUE 4 November 2017
About SAPHIR Strengthening Animal
Production and Health through
the Immune
Response” (SAPHIR) is a €10.7
million project funded by the EU
and the Swiss Government.
SAPHIR brings together
interdisciplinary expertise from
14 academic institutes including
a Chinese partner, 5 SMEs, 1
pharmaceutical company and
EFFAB.
SAPHIR is a H2020 project
running from 1 March 2015 to 28
February 2019.
SAPHIR 1
NEWS 2
PROFILES 12
EVENTS 13
PUBLICATIONS 14
CONTACT 14
SAPHIR: Through Innovative Approach!
NEWSLETTER
2
NOVEMBER 2017 - ISSUE 4
(BRSV) and Mycoplasma bovis vaccines for cattle were
presented by relevant WP Leaders from different European
Research and Knowledge Institutes.
It was decided to increase the number of researchers who
would be supported by the Young Scientist Program (YSP).
The third and last call for the SAPHIR YSP for 2018 (exactly
Dec 2017-Feb 2019) is already launched. This program
proposes financial support to young scientists to attend
conferences where they disseminate SAPHIR results. The
applications will be evaluated and the last round of SAPHIR
Young Scientists will be selected in November 2017.
Next Annual Meeting will be organized in Edinburgh, UK
by Roslin Institute. The SAPHIR Annual Meeting will take
place following the Veterinary Vaccinology Workshop
organized in cooperation with the Veterinary Vaccinology
Network and PARAGONE.
NEWS
2nd Annual Meeting of SAPHIR has taken place
between June 7-9, 2017 in Lelystad, the Netherlands.
The meeting was hosted by SAPHIR Partner
Wageningen Bioveterinary Research (CVI).
The meeting was opened by Isabelle Schwartz,
SAPHIR Project Coordinator. A brief summary of the
project progress was presented by Isabelle in which
an overview was given about the submitted 35
deliverables, and published 12 oral/poster
communications and 4 papers.
Following the opening the director of CVI, Ludo J.
Hellebrekers, welcomed the participants and
presented the Wageningen Bioveterinary Research.
During the presentations of different Work
Packages, results of some researches were shared
with the partners. In
particular the progress in
the researches on Porcine
Reproductive and
Respiratory Syndrome
Virus (PRRSV) and
Mycoplasma
hyopneumoniae vaccines
in pigs, Eimeria and
Clostridium perfringens
vaccines in poultry, Bovine
Respiratory Syncytial Virus
SAPHIR Annual Meeting 2017
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stewardship programs and alternatives to the continued
reliance on antibiotics in agricultural production need to be
developed.
The ATA symposium focused on five product categories
that could reduce the use of medically important antibiotics
in animal health and production: (1) vaccines, (2) microbial-
products and (5) innovative drugs, chemicals and enzymes.
The SAPHIR project aims at developing safe vaccines for
animal production against a selection of frequently
occurring pathogens, which blends perfectly in the first
category covered by the ATA symposium. Recently, the
World Organisation for Animal Health (OIE) has prioritized
the chicken, pig and fish diseases for which development or
improvement of vaccines could reduce antimicrobial use in
animals (Table 1,2). The diseases were ranked by animal
group, which cause the highest use of antimicrobials in the
animal. The OIE has given a high vaccine research priority
YOUNG SCIENTIST PROGRAM
NEWS
In December 2016, the 2nd
International Symposium on
Alternatives to Antibiotics
(ATA) was held in Paris.
The symposium focused on the latest scientific
breakthroughs and technologies that provide new
options and alternative strategies for preventing and
treating diseases in animals and reducing the use of
medically important antibiotics in agriculture. The
global increase in antibiotic resistance among
bacterial pathogens is believed to be due to the over-
and misuse of antibiotics in human and animal health
and agriculture. One of the key public health
concerns linked to agriculture is the potential
development of antibiotic resistant strains within
food animal production facilities and among food-
borne bacteria that could seriously compromise
therapeutic options and medical interventions. Thus,
Highlights of the “Alternatives to Antibiotics Symposium” By Evy Goossens - Ghent University, BELGIUM
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YOUNG SCIENTIST PROGRAM NEWS
or other activation receptors on epithelial cells, antigen
presenting cells and/or lymphocytes to sufficiently stimulate
the mucosal immune system. Another strategy to mount an
efficient intestinal immune response is by using Bacillus
spores as a delivery system for heterologous antigens.
Currently an oral spore vaccine to Clostridium difficile
infection in humans is under phase 1 evaluation in Germany.
Additionally, a spore vaccine to C. perfringens infections in
poultry is being developed in the SAPHIR project (in
collaboration with Prof. S. Cutting, SporeGen and Prof. F.
Van Immerseel, Ghent University).
The first results of this collaboration were presented at the
symposium. In addition to the use of novel adjuvants and
Bacillus spores for induction of mucosal immunity, also other
recombinant oral vaccines platforms, such as Salmonella-
vectored subunit vaccines, were presented at the ATA
symposium (L.R. Bielke, Ohio State University). Moreover,
the use of highly conserved antigens to provide cross-
protection among multiple species of pathogens was
explored.
to necrotic enteritis (caused by Clostridium
perfringens) and coccidiosis (caused by Eimeria) in
broilers and PRRSV in swine, underlining the
importance of the SAPHIR project.
One of the challenges in developing efficient
vaccines against enteric pathogens is to mount a
good mucosal immune response. To induce antigen-
specific immunity, a vaccine has to pass protective
barriers and to overcome innate defense and
tolerance mechanisms before it can activate the gut-
associated lymphoid tissue (GALT). More specifically,
a vaccine has to reach the epithelial close to the
GALT in an immunogenic form, pass the epithelial
barrier to reach cells which can present the antigen to
lymphocytes of the GALT, and induce an antigen-
specific immune response which can prevent the
pathogen to cause disease.
The research group of Prof. E. Cox (Ghent
University) is currently investigating innovative
adjuvants which target pattern recognition receptors,
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gave a detailed lecture about the genetic exchange and inflammasome sensors activated by cyst-forming parasites (Toxoplasma, Neospora and Sarcocystis). Sub-plenary talks were mainly dedicated to Toxoplasma (life cycle evolution and transcriptional profiling during mice infections), Eimeria (description of cryptic species, by SAPHIR member Damer Blake) and Cryptosporidium parasites (genomic studies applied to therapeutics).
The Vaccines and therapeutics session was divided in two main pillars: the discovery of new drugs against toxoplasmosis by targeting host metabolic pathways (Silvia Moreno, University of Georgia), and the development of vaccines based on Eimeria-recombinant vectors (Suo Xun, College of Veterinary Medicine, China Agricultural University). This session was focused on the development of control measures against Eimeria parasites of poultry. During one of the sub-plenary sessions, I had the opportunity to present the positive results we have obtained under the SAPHIR project using transgenic Eimeria parasites as vaccines.
The last session of the ICC covered the topic of Host-pathogen interactions, primarily focusing on poultry coccidiosis. Woo H. Kim and Hyun S. Lillehoj (Animal Biosciences and Biotechnology Laboratory, USDA-ARS) discussed several aspects of the host immune response to avian coccidiosis, and SAPHIR member Marie-Hélène Pinard-van der Laan (Institut National de la Recherche Agronomique) described breeding and genetics strategies to improve host resistance to coccidiosis.
Thanks to its excellent organization, participation in the ICC/ASP conference provided a great opportunity to disseminate information on our SAPHIR project with the international scientific community and allowed networking with potential industry partners.
NEWS
12th International Coccidiosis Conference and 92nd Annual Meeting of the American Society of Parasitologists
By Iván Pastor-Fernández - RVC, UK
Parasitologist’s were in luck during 2017, when the 12th International Coccidiosis Conference (ICC) and the 92nd Annual Meeting of the American Society of Parasitologists (ASP) were merged into a
single event, held in San Antonio, Texas, from the 27th of June to the 1st of July.
The ICC was organized in four sessions:
(I) Systematics, taxonomy and evolutionary biology; (II) Genetics and comparative genomics; (III) Vaccines and therapeutics; and (IV) Host-pathogen interactions.
The ASP meeting covered similar topics, but with a particular focus on evolutionary ecology and epidemiology of parasites affecting wild and domestic animals.
The first ICC session was dedicated to Systematics, taxonomy and evolutionary biology, and included two plenary talks focused on the population structure and host specificity of Eimeria affecting wild animals (Jana Kvičerová, University of South Bohemia) and Cryptosporidium parasites affecting humans and animals (Lihua Xiao, Centers for Disease Control and Prevention). Four sub-plenary talks covered similar topics in the related parasites Toxoplasma gondii and Cycloscora cayetanensis.
During the session Genetics and comparative genomics, Boris Striepen (University of Georgia) focused his plenary talk on the molecular genetics of Cryptosporidum, while Michael E. Grigg (National Institute of Allergy and Infectious Diseases, USA)
YOUNG SCIENTIST PROGRAM
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pathogens and trigger an immediate response limiting the
further spread of the pathogen. This innate immune
response is initiated by receptors recognition receptors
(PRR) specific to different classes of pathogens, which are
called pathogen-associated molecular patterns (PAMPS).
The innate immune system can protect the body from
infection, reduce the ability of a pathogen to proliferate and
spread in the host, and importantly trigger adaptive
immune responses. With respect to the latter, it will direct
the type of adaptive immune response, for instance
promote the response required against extracellular
pathogens colonizing mucosal surfaces or alternatively
promote responses against intracellular pathogens.
Basic research has mostly used the mouse model to
discover fascinating details of the functioning of the
WORK IN PROGRESS
NEWS
WP9- Translating basic veterinary immunology to innovative vaccines By Artur Summerfield, Gael Auray and Roman Braun - IVI, Switzerland
INTRODUCTION
Infectious diseases are a major cause for animal
suffering and production losses in livestock and
sometimes they also present a serious hazard for
food safety. Combating infectious diseases of
livestock is therefore a top priority. Mass use of
antibiotics, antiparasitic drugs and vaccines have
been employed, but this practice alone was unable to
provide solutions to many of the multifactorial
disease complexes. The increasing thread of
multidrug resistance caused by inappropriate
employment of antibiotics is a serious concern.
Although vaccines are a great success story they are
lacking efficacy against difficult pathogens and
against certain disease complexes caused by multiple
pathogens typically found in the field.
The mammalian immune system has evolved to
control pathogenic infections and in most cases, is
able to perform its tasks with remarkable efficacy.
The immense variety of pathogens has driven the
evolution of a very complex immune system, which
nevertheless follows relatively simple principles. The
immune system needs to recognize invading
Figure 1: One of the approaches of Workpackage 9 of SAPHIR for improved vaccines.
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BRIDGING THE GAP
Veterinary immunology focuses on the immune system of
the target species of pathogens and is dedicated to
understand the functioning of immunity in pigs, cattle,
small ruminants, chicken and other relevant species. This
research has demonstrated important species-specific
differences in the immune system. In the frame of SAPHIR
we have analysed in detail the biology of antigen presenting
cells, including dendritic cells and B cells in the pig,
presented in more detail below. These cells are central for
the induction of adaptive immune responses and therefore
represent excellent targets for immunostimulation or
immunomodulation to improve vaccines. An important part
of this work was also the identification of ligands which are
particular strong at activating specific cell types. Such
ligands are currently being tested in vaccine formulations of
the SAPHIR project (Figure 1).
WORK IN PROGRESS
NEWS
mammalian immune system. But the amount of new
prophylactic and therapeutics solutions to human
and animal health has been disappointing. While the
murine model is excellent to discover and explain
basic principles of the mammalian immune system,
details in the distribution and functioning of certain
immune cells and immunological receptors and their
ligands can differ. These differences have been driven
by evolution as different species habitat a different
environment with for instance different food and
pathogen exposure. In addition, major anatomical
differences, for example in the skin, the mucosa, and
the gastrointestinal tract, have a strong influence on
the functioning of the immune system. As a
consequence, extrapolations of adjuvant research
performed in the murine model to other species has
often been disappointing.
Figure 2: DC subsets, their ontogeny and main functions
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response of porcine DC subsets to Toll-like receptor (TLR)
ligands, representing a class of PRR, revealed peculiarities
of the porcine immune system. In particular, we found that
pDCs are by far the most important source of TNF-α, IL-
12p40, and of course IFN-α, while cDCs are most efficient in
MHC and costimulatory molecules expression. On one side,
our data demonstrate that extrapolation of data on DC
biology from one species to another has to be done with
care, on the other side it shows how functional details have
differentially evolved in different species. The knowledge
acquired is now employed for the design of new adjuvant
formulations in the frame of SAPHIR.
PORCINE B CELL SUBSET AND THEIR
RESPONSES TO PAMPS
Considering that TLR triggering of B cells are known to
promote B cell expansion, differentiation of B cells into
antibody-producing and memory cells, these pathways are
attractive to improve animal vaccines. Our study published
in Frontiers in Immunology (10.3389/fimmu.2017.01044)
investigated the response pattern of porcine B cell subsets
to a large collection of TLR ligands and identified ligands
that are particularly efficient at activating B cells. We also
identified differences in the response of various B-cell
subsets in the response to such ligands and describe for the
first time innate-like B subsets representing the mains
source of immunoglobulin M following TLR stimulation.
WORK IN PROGRESS
NEWS
PORCINE DENDRITIC CELL SUBSET AND
THEIR RESPONSES TO PAMPS
Dendritic cells (DCs) represent a family of
professional antigen presenting cells which are not
only essential for the induction of adaptive immune
responses but also provide an important link between
innate and adaptive immune response, and shape the
immune response depending on the innate signals
received and the exposed tissue microenvironment.
Using a large panel of antibodies in multicolor flow
cytometry, cell sorting and RNA sequencing, our
work published in The Journal of Immunology
(10.4049/jimmunol.1600672) has identified and
characterized in detail porcine plasmacytoid DCs
(pDCs) and conventional DCs (cDCs), the latter being
further divided into a cDC1 subset and a cDC2 subset.
While pDCs are particularly important for anti-viral
responses through production of high amounts of
type I interferons (especially IFN-α), cDCs are most
efficient at presenting antigen and activating naïve T
cells. cDC1 are specialized in stimulating CD8 T cells,
whereas cDC2 have been shown to be efficient at
promoting Th17 as well as Th2 immune responses
(Figure 2).
Despite cross-species conserved subset-specific
transcripts, porcine DC differed from the species
described so far in many expressed genes. The
Stimulating such
responses to
improve animal
health might have
a great potential
to improve
vaccines and
identify
alternatives to
antibiotics.
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NOVEMBER 2017 - ISSUE 4
SAPHIR WP16 uses mathematical modelling and statistical
inference to develop predictive models to assess
epidemiological and evolutionary consequences of
vaccination for livestock diseases such as PRRS. The results
of these models can help researchers and vaccine producers
within and outside SAPHIR to develop safer and more
effective vaccines, and farmers and policy makers to
develop vaccination programmes that keep pig farms PRRS
free long-term.
One of the big challenges in vaccine development and
infection control is that not all animals respond the same
way to infection or vaccination, and that the response is
WORK IN PROGRESS
NEWS
WP16- How to prevent viremia rebound in virus infections? Evidence from a data-driven mathematical model for PRRS
By Andrea Doeschl-Wilson , Natacha Go and Suzanne Touzeau
Vaccines can drastically alter the environment in
which pathogens live. Indeed, the goal of vaccination
is not only to protect individuals from disease, but to
decrease the risk and severity of disease outbreaks in
populations, and ultimately to eradicate the disease.
For the porcine reproductive respiratory syndrome
(PRRS), one of the most devastating pig diseases in
the world and endemic in most European countries,
vaccination has so far proven ineffective in
controlling the disease. Indeed, the fast evolving,
highly genetically diverse PRRS virus poses a real
challenge for developing fully protective vaccines
with long-term effectiveness.
Figure 1: Functional diagram of the model representing the within-host immune response to PRRSv infection.
Binding of PRRS viral particles (V) and naive target cells (APC) either result in mature and non-infected cells (APCm) that phagocyte viral particles, or in mature and infected cells (APCi) that allow viral replication and excretion of new viral particles. Viral particles are neutralised by antibodies (Ig); infected cells are cytolised by natural killers (NK) and cytotoxic lymphocytes (CD8+). Mature target cells (ACPm and ACPi) present the viral antigen to naive adaptive effectors (CD4+) which activate the CD8+ and B lymphocytes (B). B cells synthesise antibodies. Cytokines are synthesised by various effectors ( ACPm, ACPi, NK, CD4+, CD8+) and regulate most of the immune mechanisms (phagocytosis, infection, excretion, antigen presentation, cytolysis). Regulations are either activation, amplification or inhibitions.
Colours – green: PRRSv particles; red: innate response; blue: adaptive response; purple: both innate and adaptive responses.
Lines – plain with arrow: state changes; dashed (dotted) with arrow: (cytokine) syntheses; plain dark grey with bullet: regulations by cytokines.
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profiles but also offers, for the first time, insight into
potential causative immune mechanisms for generating
rebound. In particular, contrary to current hypotheses
emerging from genetic analyses, this model reveals that
viremia rebound can occur as a result of between-host
differences in the immune competence alone, without the
commonly hypothesized emergence of viral escape
mutants.
Amongst the identified candidate immune mechanisms,
the data-informed model revealed that rebound is
promoted by high target cell apoptosis, high cell infection
and low cytolysis of infected cells by cytotoxic lymphocytes,
while increasing virus neutralisation efficiently prevents
rebound. The model results suggest that vaccines or
genetic selection promoting strong neutralising and
cytolytic responses, ideally associated with low apoptotic
activity and cell permissiveness, would prevent rebound.
Cytotoxic lymphocytes and neutralising antibodies are
usual targets for vaccine development. However, given the
WORK IN PROGRESS
NEWS
strain-dependent. In particular, virus load rebound
following a steady phase of viral decline (see Figure 2)
is a common and highly undesirable phenomenon for
PRRSv and other viral infections across a range of
species. These rebounders not only suffer prolonged
infection themselves, but are also likely to maintain
the disease in the herd for longer. What causes some
individuals to experience viremia rebound while
others manage to steadily clear the virus?
To determine whether rebound can be caused by
differences in the immune response alone,
researchers from INRA, Inria, France and The Roslin
Institute, UK created a mathematical model of PRRS
within host infection dynamics (see Figure 1) and
fitted it to a dataset from a large scale experiment
(see Figure 2), in which thousands of genetically
diverse pigs were challenged with a virulent PRRS
virus strain (courtesy of the PRRS Host Genetics
Consortium). This mechanistic infection model, fitted
to the experimental data, not only successfully
reproduced the observed wide range of viremia
Figure 2: Modelled viremia profiles compared with the data (lower and upper envelope: black curves) for the A. non-rebound, uniphasic profiles (green) and B. rebound, biphasic (red) profiles.
Black boxes: data ranges for the first viral peak, the rebound peak (max) and the minimum between the two peaks (min). Dashed line: viremia detection threshold. Semi-log graphs.
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The research was carried out as a collaboration between
SAPHIR, MIHMES, an INRA led multi-scale modelling
project (https://www6.inra.fr/mihmes), and the PRRS Host
Genetics Consortium (PHGC). The authors would like to
thank the SAPHIR coordinator Isabelle Schwartz for her
constructive comments to this article.
WORK IN PROGRESS
NEWS
high diversity of circulating PRRSv strains, cross-
protection remains a major challenge for PRRSv
vaccination. Consequently, alternative solutions that
target non-specific immunity, in particular those
leading to lower target cell permissiveness and/or
reduced apoptotic activities are particularly relevant.
Non-specific immunity can originate from host
intrinsic, genetically driven, innate immune
responses or from alternatively trained immunity.
This non-specific alternatively trained immunity
could result from attenuated vaccines against other
pathogens, as shown in infants, or could be explicitly
elicited by immunostimulants such as those indicated
by the model (such as TNFα) or others yet to be
defined.
The findings of this theoretical study have profound
consequences for the development of successful
PRRS intervention strategies, as they would imply
that rebound and thus possible prolonged virus
transmission in a pig herd can be prevented by
altering the immune response through vaccines or
other pharmaceuticals. The identified immune
mechanisms for preventing rebound could also help
to identify pig genes associated with prolonged virus
By Roman Othmar Braun, Sylvie Python, Artur Summerfield
(FrontImmunol, 2017)
This project has received funding from the European Union’s Horizon 2020 Programme for research, technological development and demonstration under the Grant Agreement n°633184 and Swiss Government.
This publication reflects the views only of the author, and not the European Commission (EC). The EC is not liable for any use that may be made of the information contained herein.