Author: rob

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A network of labs that study and survey Tick-Borne Flaviviruses

The best results stem from collaborative efforts of a great team of experts across nations. When the work of the virologists involved in the Tick-Borne FlaviVirus – TBFVnet project, was affected by the Covid-19 pandemic, and later, when we lost our Russian partners due to the sanctions imposed following the outbreak of the war with the Ukraine, the determination to network, nonetheless ensured the value and integrity of their findings.

TBFVnet established a network of laboratories sharing best practices for detection and surveillance of tick-borne flaviviruses, a class of viruses delivered by ticks which are endemic in eastern and central Europe but moving west. The network wanted to be able to detect the infection, but also to develop antiviral drugs, which were not yet available, and to expand to other regions impacted by the disease, such as Slovenia and the Republic of Moldovia. During the four years of the project, the network published results in scientific journals, and presented these at meetings for both the scientific and lay public. It optimised protocols and key reagents for virus detection, at the molecular and blood serum level, and completed whole virus genome sequencing, which was shared between the laboratories.

The ramifications of education and training in laboratory techniques also through an exchange programme of junior staff between the laboratories in the different countries of the network (Italy, Czech Republic, Slovakia, Norway), has contributed to the establishment of a common strategy on TBFV pathogenesis studies and antiviral drug discovery. The exchange of researchers also contributed to their career development, as doctoral or master students, providing exposure to a different, international environment.

The project has involved tens of researchers, scientists, and students, and a considerable number of the public (200 people live and online attending the public engagement event on the topic; to date explanatory videos on YouTube have reached 1,000 views). The opportunity given to young women in science in the context of the project, to learn and build their own networks for career development, signifies perhaps one of the greatest impacts achieved.

Consolidated and clear media coverage of all these results, particularly at a time when disinformation and the disintegration of the social and democratic fabric is a real threat, undoubtedly constitute some of our best practices.

Most importantly, communications were based on sound scientific knowledge, and prepared by experienced communicators versed in the latest techniques and developments in science dissemination. Thanks to dynamic visuals, regular updates, a broad readership, and the support of the Regional Cooperation Magazine in carrying over some of the best stories, we look forward to the impacts created by the project continuing well into the future.

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Recent outbreaks of viruses such as Chikungunya, Zika, Yellow Fever, West Nile Fever, or Dengue, as well as the SARS-CoV-2 pandemic, have found us unprepared and vulnerable.

The spread of these viral pathogens is determined by complex demographic, environmental and social factors, causing diseases to emerge in countries where they were previously unknown. Coupling diagnostics and epidemiology with digital platforms for processing data from various sources leads to the possibility of an open, global and digital pathogen surveillance system with predictive capabilities.

The TBFVnet project, focusing on surveillance and research on Tick-borne flaviviruses (TBFV) encompasses topics relating to public health and raising awareness. With a view to increasing knowledge, facilitating information, and influencing attitudes and behaviour, as well as to further disseminate and promote the objectives of our project, we are holding a public engagement event for the lay public.

Dr. Alessandro Marcello (ICGEB), project partner, will be participating with invited speaker, Dr. Marta Giovanetti, Campus Bio-Medico University of Rome (UCBM), and O. Cruz Foundation, Rio de Janeiro, Brazil, in the event entitled: Are we more exposed to new viral outbreaks? Addressing the risk with a one-health approach. The event, under the “Science & the City” umbrella of public scientific conferences, will be held at 3 pm on Sunday 24 September 2023, in the Centre of Trieste, during the Trieste Next Festival of Scientific Research. In its 12th edition, the festival attracts thousands of participants each year and constitutes a great opportunity to communicate the results and benefits of scientific research to the lay public.

Entrance is free and the event will also be streamed live. The event will be in Italian for the local populace, however, a podcast and short film will be produced afterwards with English subtitles and distributed on YouTube. Additional information about the event and the project will be available at the ICGEB stand in the main square of Trieste, Piazza Unita’ d’Italia throughout the festival, from Friday 22 to Sunday 24 September.

Dr. Fabio De Pascale, Science Communicator, University of Padua, Italy, Communications Consultant for the TBFVnet project for two years, will moderate the talks.

An ongoing theme for our scientists concern viruses invading new areas, representing risks and the need to be monitored. The focus of the TBFVnet project and its relevance to the monitoring of novel viral species will also be discussed.

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In a series of short videos, we hear directly from our partners and the roles of their institutes and research teams on the TBFVnet project.

Daniel Ruzek, PhD, Associate Professor and Principal Investigator at the Veterinary Research Insitute (VRI) in Brno, Czech Republic, is lead partner of the network. In this brief video he talks about the purpose, scope and some of the milestones of the TBFVnet project. He explains how this is important for international collaboration and introduces the work of the VRI. The Czech Republic is at the heart of the endemic area for TBFV, between Western Europe and the East. The research of the Veterinary Research Institute focuses on the pathogenesis of TBE, development and testing vaccines and antivirals and is also involved in national and international surveillance and diagnostic programs. Therefore, the Veterinary Research Institute is best placed to coordinate the activities of TBFVnet.

Boris Klempa, PhD, Head of Virus Ecology Group, and Principal Investigator at the Biomedical Research Center of the Slovak Academy of Sciences, partner in the network, introduces the work of his institute and his Group’s study of the interaction between tick-borne encephalitis virus (TBEV) and the tick. He describes how this impacts a One Health approach. The Biomedical Research Center has long-standing expertise in a TBEV-tick-model. This expertise allows characterisation of new virus isolates in terms of their tick replication and transmission efficiency.

Ashild Kristine Andreasson, Senior Scientist and Principal Investigator at the Norwegian Institute of Public Health NIPH, introduces her institute and tells us about TBEV, which animals transmit the virus, and the numbers of TBFV infections in Europe and worldwide. She also explains the most typical symptoms, and defines encephalitis. Further, she explains why is it underestimated, and what the scientific community (and the TBFVnet team) can do to improve the surveillance and diagnosis of TBEV and other TickBorne FlaviViruses. Dr. Andreasson touches upon the broader impact of the project to similar studies in this field. NIPH is sequencing and analyzing the genome of TBE isolated viruses in relation to their inflammation potential to the central nervous system. This work will bring new knowledge to improve the diagnostic potential of encephalitis in humans.

Alessandro Marcello, PhD, is Group Leader of the Molecular Virology Laboratory and Principal Investigator at the International Centre for Genetic Engineering and Biotechnology, ICGEB in Trieste, Italy. He tells us about the role of ICGEB in this project, the European Economic Area and the Norway Fund. He also explains why it is important to sustain international cooperation in science, and a One Health approach, revealing how this project might be pivotal in finding new therapies or strategies to prevent infection. The Molecular Virology Laboratory in Trieste, Italy studies human pathogenic viruses. Within this project the lab is involved in developing viral diagnostic methods and studies the host cell response to TBEV infection to discover antiviral drugs. The ICGEB has the expertise to promote, facilitate, increment and assist international networking and communication.

Olga Sofronie is a PhD student in the Molecular Virology laboratory at the ICGEB in Trieste, Italy. She tells us her background and what has fuelled her passion for science, by discussing the aim of her project. Since the end of January 2023, she has worked to set up a molecular diagnostic method for TBEV detection. Her involvement in the TBFVnet project at ICGEB Trieste has been predominantly to become familiar with state-of-the-art techniques, equipment, and methodologies. She explains how this has consolidated her research. We also ask about her vision for the future. Her visit has been funded through the EEA and Norway Grants Fund for Regional Cooperation as part of the Tick-Borne Flavivirus network project.

Adam Kevely, PhD student from SAS, Slovakia, spent one month in the Molecular Virology Laboratory at the ICGEB, in Trieste, Italy studying high-throughput screening techniques for antiviral drugs discovery. He also collaborated with the laboratory of Daniel Ruzek, at the VRI in Brno, in the Czech Republic. The results of this collaboration are part of Adam Kevely’s first author scientific paper. Here, he recounts how his involvement in the TBFVnet project has advanced his scientific knowledge and career. He also considers how the possibility to collaborate with other Groups in different countries enriches scientific experience.

Link to the Video interviews with TBFVnet partners on YouTube.

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The second TBFVnet regional seminar was held at the International Centre for Genetic Engineering and Biotechnology (ICGEB) in Trieste, Italy on 26-28 March 2023. The TBFVnet project, led by the Veterinary Research Institute (VRI) in Brno, brings together researchers from the Biomedical Research Center of the Slovak Academy of Science (SAS) in Bratislava, The Norwegian Institute of Public Health (NIPH), and the ICGEB in Trieste, Italy (www.icgeb.org). In addition, researchers from associated laboratories from Sweden, Slovenia, Russia and Moldova are part of the enlarged area of interest of TBFVnet.

The Workshop was attended by members of the network with additional experts to provide updates on the progress of the project and to learn about novel developments in research, diagnostics and surveillance. Organised by Alessandro Marcello and Tea Carletti (ICGEB Trieste, Italy), and by Åshild K. Andressen (NIPH), approximately 40 participants attended the two-day meeting, including members of the network, collaborators and invited speakers from the Italian Regional University Health Authority (ASUITS), the Italian Higher Institute of Health (ISS), and experts from the University of Ljubljana, Slovenia, Umeå  University, Sweden, and the University of Cambridge, UK.

Bringing together collective expertise, sessions focused on diagnostics and surveillance of Flaviviruses and the importance of reference laboratories for the diagnosis of emerging and re-emerging viral infections. Other topics discussed included Tick-borne encephalitis in domestic animals, vaccine development, infectivity and resistance mechanisms. Among the techniques presented were ex vivo imaging and tools for nanoscale characterisation.

The effect of host genetic background on the clinical course of TBE and identification of novel biomarkers, i.e. biological molecules found in blood, other body fluids, or tissues that are a sign of, in this case, brain damage were presented by Daniel Ruzek, VRI. The interplay between the endoplasmic reticulum (ER) stress response, and the antiviral response to TBEV infection was presented, as well as a screening for host-directed antiviral therapy targeting proteostasis.

The Regional seminar provided an opportunity for each partner to present their particular expertise in a series of brief, video interviews, which are being launched to communicate the aspects of the project to the broader public.

Link to the Video interviews with TBFVnet partners on YouTube.

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The brain is perhaps the organ that most characterizes our species. It can think about itself, wonder about its origin and also compare itself with other living beings. It is our evolutionary history that led us to this point, starting from when we first gained upright stature, then the use of rudimentary tools, and then the use of language: the thinking and social interaction skills that we have today. Perhaps it is because of the role this organ plays in our species that evolution has equipped it with considerable protection. Our skull, for example, is strong enough to protect it from many shocks. There is also a barrier that carefully separates it from the rest of our body, the blood-brain barrier. Yet, this fascinating organ is not immune to disease or damage. Sometimes these affect people’s mental health, while others affect the neurological system itself and can also come whence one would least expect. 

Neurological disorders are a class of diseases triggered by dysfunction in the cells of the central and peripheral nervous system. In some cases, such as in Alzheimer’s or Parkinson’s, these dysfunctions also affect the cognitive abilities of patients. In others – such as epilepsy, multiple sclerosis, or amyotrophic lateral sclerosis – it is mainly the motor abilities that are affected while the cognitive ones remain almost intact. The causes of many neurological disorders are still unclear and, for the most part, many different factors contribute to trigger these. In some cases, it seems that viruses and the responses they trigger in our body also play an important role. 

The repercussions of lockdown due to the COVID-19 pandemic on our mental well-being have often been discussed. However, little is known about the direct effects of the SARS-CoV-2 infection on the nervous system. There have been numerous cases of people infected with COVID-19 who had also developed both mild neurological consequences, such as confusion and agitation, but also severe consequences, such as dysfunctions of the corticospinal apparatus, which controls our voluntary movements, and loss of consciousness. It is unclear whether these were due to the prolonged inflammation generated by the body in response to viral infection or because of direct entry of the virus into the nervous system. SARS-CoV-2, however, is not the only coronavirus to have consequences on the central nervous system. SARS-CoV-1, the first virus to trigger the SARS epidemic in the early 2000s had both immediate and long-term impacts on the central nervous system. The same is also true for MERS-CoV, another very lethal and dangerous coronavirus that has remained, fortunately, contained. How do viruses that attack the respiratory tract also affect the central nervous system? 

Our brain is well protected in our head, the skull protects it from shocks and the blood-brain barrier blocks all foreign agents that can come from within our own organism; or at least it tries to do so. This barrier is made up of a set of cells that are highly specialized in allowing only certain substances to pass from the blood to the brain. Anything that is in the blood that is not recognized by these cells can pass. Sometimes, however, something manages to overcome this defense. Pathogens can reach the brain either through what is called a Trojan horse, i.e. by infecting cells of the immune system that are recognized as “friendly” by the blood-brain barrier and allowed to pass, or by going up through the axons, the connections that link nerve cells spread throughout the body with the brain. This is exactly how viruses can travel up the olfactory system and from the nose arrive, undisturbed, to infect the central nervous system. 

It is not, however, only respiratory viruses that create neurological problems. Some prolonged viral infections can trigger neurological damage. HIV infection, for example, can have neurological consequences similar to Parkinson’s in some cases, as can herpesvirus, such as chickenpox and some hepatitis that can cause neurological damage in the long term. There are also viruses that are contracted through animals such as the tick-borne encephalitis virus (TBEV), which has direct consequences on the brain. TBEV, despite being a virus that infects humans through a small puncture on the skin, causes a powerful inflammation of the brain that can lead to severe headache, loss of consciousness, seizures, and difficulty in speaking and hearing. How it gets from skin to the brain is not yet completely clear, but one likely route is the “Trojan horse”, infecting cells of the immune system. 

Apart from a few well-known cases, a clear correlation between viral infections and neurological and neurodegenerative diseases has not yet been demonstrated. However, given current knowledge, it seems clear that prolonged stress on the immune system generated by certain viral infections can in some cases increase the risk of developing one of these diseases. It is not only viruses that affect the nervous system that cause problems, also those that affect other areas of the body aggressively or for a prolonged period can do so. 

Viruses existed on Earth long before humans. They found many ways to infect their hosts and bypass their defenses. We humans have many innate tools (i.e., the immune system) that combat these infections and we also have many drugs and vaccines to help us do so. The only way we can keep pace with viruses is through continued scientific research that may, one day, lead us to understand the role of viruses in neurological disorders and thereby help us to prevent them or to find a cure. 

This article was originally published in the Regional Cooperation Magazine, issue 12, in February 2023

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In 2022 partners from TBFVnet published two important research papers. The first was released in April and was the result of the work of the Molecular Virology Laboratory at ICGEB. The second was published in November as a joint effort between the Biomedical Research Center, the Slovak Academy of Sciences and the lead partner, the Veterinary Research Institute in the Czech Republic. Each paper investigates two different aspects of research on tick-borne flaviviruses.

The first study focuses on the possibility to develop a new antiviral against flaviviruses. Antivirals are a specific class of drugs useful to treat viral diseases. They can only be used within the very first hours at the onset of the infection as later they start losing their effectiveness. Compared to the number of viruses that can infect humans, very few of these drugs are available, among which those against prominent endemic viruses such as influenza, HIV, hepatitis C and herpesviruses.

Antiviral drugs do not kill the virus as some antibiotics do with bacteria. Usually, they act by impairing virus replication within the cell or its ability to infect other cells. This is why they need to be given to the patient at the onset of infection: the sooner they act, the earlier they can block the spread of the virus in the body. But this is also the reason why antivirals are few and difficult to develop. To replicate and proliferate viruses use the host cellular machinery, the same mechanisms that are present in healthy cells. Thus, if antivirals are not carefully developed, they can be toxic and harmful to the human body. But there is another possibility, at least against one class of viruses.

Most viruses have a closed protein structure containing their genetic material, but some share a peculiar lipidic envelope surrounding the core. It is through this membrane that these viruses enter the cell and infect them. This is a characteristic of all flaviviruses, including those carried by ticks. A drug targeting this envelope, or any of the steps in which it is involved, could represent a precious tool. It could serve as a broad treatment for a whole class of viruses while we wait for vaccines or more specific antivirals to be developed. However, in the most promising scenario, they can also be used as a first-line defence against emerging infections.

ICGEB researchers explored the role of recently synthetic chemical compounds, derived from a molecule called pyridobenzothiazolone, in reducing viral infection by West Nile virus, a flavivirus that represents a serious threat to human health. The study, published in the journal Viruses, confirmed the role of this compound in inhibiting infection from the virus. Although the mechanism of action is not fully elucidated, this research confirms that these chemical compounds can act on the virus envelope. This opens the way to the development of antiviral drugs against all flaviviruses including those carried by ticks such as tick-borne encephalitis virus.

The second research project presented a new tool to investigate the interaction between a virus and its vector deeply. In particular, the authors from the TBFVnet partner institutions, the Biomedical Research Center and the Veterinary Research Institute, experimented with a new procedure to study the interaction between tick-borne encephalitis virus (TBEV) and the tick. In fact, very little is known about how these two organisms live and share their life cycle. TBEV can live within the ticks without harming the tick in any way and from there infect humans and animals. This partnership is, indeed, at the basis of the cycle that transmits the virus to all its hosts including humans. Understanding how the virus lives and replicates in ticks and is then transmitted to the hosts could be pivotal in finding new therapies or strategies to prevent infection.

The researchers used an engineered version of TBEV that carries the mCherry reporter gene and tests its efficacy to study different stages of tick and virus interaction. A reporter gene can in fact shed light on specific molecular mechanisms; it can be seen through microscopy as it emits light when stimulated and researchers can thus monitor the virus’ presence in any experiment. This study confirmed that the TBEV mCherry is a valuable tool for studying the biological mechanisms between the virus and the tick. With this new tool, it could be possible to better understand the biology of TBEV and, in future, find new ways to block its infection.

These two studies represent precious milestones for the TBFVnet project. The research that can be done in this field can effectively give new solutions to the threat that flaviviruses carried by ticks represent to human health.

The research papers cited in the article can be found at the following links:

• Virucidal Activity of the Pyridobenzothiazolone Derivative HeE1-17Y against Enveloped RNA Viruses. Bonotto et al., Viruses 2022 doi: 10.3390/v14061157
• Fitness of mCherry Reporter Tick-Borne Encephalitis Virus in Tick Experimental Models. Kevély et al., Viruses 2022, doi: 10.3390/v14122673

Each year, young scientists from almost 70 Member States undertake research in the laboratories of the International Centre for Genetic Engineering and Biotechnology. Youth, essential to nurturing a brighter and more equitable future, constitutes the backbone of the ICGEB’s training programmes that have been operating for over three decades across the world.

To celebrate the month of the European Youth Goals, ICGEB hears from some of the Fellows from across Europe who are undertaking research in the laboratories in Trieste, Italy. Their diversity, wealth of experience, and passion for science are testimony to the importance of project funding. Among these are Fellows working in the Molecular Virology laboratory where the TBFVnet: surveillance and research on tick-borne aviviruses (TBFV)project of the Norway Grants Fund is carried out.

Denis Rajnović was born and spent part of his childhood in Sweden, after which his family returned to Rijeka (Fiume), Croatia. “I finished my Bachelor and Master’s degree in drug Research and Development at the University of Rijeka. During my Master’s, I performed internships in recombinant protein production, recombinant protein interaction analysis and yeast genetics, in Slovenia and Spain. While pursuing my PhD degree in Microbiology at the Autonomous University of Barcelona, I also studied influenza biosensors, as well as in single cell manipulations with FluidFM technology in Zurich, Switzerland.” Under an ICGEB-Elettra fellowship Denis pursues research in structural biology, biophysics, life sciences, and biotechnology to elucidate biological and behavioral characteristics of SARS-CoV-2 variants and to contribute to pandemic preparedness.

Chiara Kalebić, also from Croatia, completed her Master’s degree in Functional Genomics at the University of Trieste and in Molecular and Cellular Biology at the University of Rennes1, France. She was awarded a Fellowship in the Molecular Virology Group and subsequently obtained a PhD position in autumn.

Dr. Ljiljana Sjekloća, Postdoctoral fellow in the Molecular Pathology lab, hails from Montenegro. She left her home country at the age of eighteen, driven by the war of the 1990s in former Yugoslavia. “I came to Trieste because of its proximity to Montenegro and for the excellent organisation of scientific courses. I graduated in Molecular Biology and obtained a PhD in structural and functional genomics from the International School for Advanced Studies, SISSA, in Trieste, carrying out research on cytoskeletal proteins at the Elettra Synchrotron.” While a doctoral student at the University of Vienna, Ljiljana became passionate about RNA, an experience that led her to investigate the interactions between proteins and ribonucleic acids, and led to her working in numerous centres of excellence in the field, including the MRC in London, EMBL in Grenoble, the Karolinska Institute in Stockholm, Harvard Medical School, Boston, and NIH Bethesda, to become a specialist in protein and RNA modifications. She is carrying out a research project on amyotrophic lateral sclerosis (ALS), in the Molecular Pathology lab headed by Emanuele Buratti.

Emanuele Orsini, is one of the Italian fellows at ICGEB Trieste who joined the Molecular Virology group to study the interactions between SARS-CoV-2 and the host cell. “The family of Coronaviruses includes RNA viruses, causing respiratory diseases with highly epidemic potential. Indeed, the most recent epidemic outbreaks before COVID-19 were caused by coronaviruses (SARS-COV and MERS). Nonetheless, the risks posed by coronaviruses were underestimated by the research community that kept focusing on topics more relevant for the public health of western countries (such as cancer, cardiovascular or neurodegenerative diseases). The SARS-CoV-2 outbreak dramatically revealed our lack of knowledge. As an ICGEB fellow I am doing my best to give my contribution, however small, to a better understanding of coronavirus biology and to expand our toolbox against emerging diseases. At the same time, I hope the ICGEB will become a leading institution in the global community to establish research priorities.”

Urša Šušnjar Hribar, Junior Research Technician in the ICGEB Molecular Virology Lab, comes from Slovenia. Dr. Šušnjar also pursues projects in science communication and recently featured in the ICGEB documentary “The Goals of Science“, for the project on SARS-CoV-2 diagnostics. Slovenia is a partner of the TBFVnet project, as is ICGEB Member State Moldova – whose Fellow Assistant prof. Mariana Ulinici was recently interviewed for the women in science video series.

Antonio Mura, PhD student in the Molecular Medicine Laboratory at ICGEB is from Ittiri, a small town in Sardinia, where he graduated in Biotechnology. “I have lived in Trieste since 2017, when I started my Master’s degree in Neuroscience. I’m currently focusing on the characterisation of molecular pathways related to Fam3c, a protein that our laboratory discovered to protect the heart after myocardial infarction. Considering that cardiovascular diseases are the leading cause of death globally, we believe that understanding the key players involved in the beneficial effect exerted by Fam3c is of paramount importance.”

Dr. Naida Babić Jordamović is from Sarajevo, Bosnia and Herzegovina, where she worked as Research Associate at the Verlab Research Institute for Biomedical Engineering and Biotechnology. She is a postdoctoral fellow in the Computational Biology Group, where she is working under the supervision of Dr. Silvano Piazza on investigating biological pathways as biomarker signatures for precise cancer treatment underlaying the importance of personalised medicine.

ICGEB training programmes represent an exciting opportunity to interact with people all over the world in state-of-the-art scientific environments and in all the top scientific institutes. Several hundred new ICGEB fellows yearly constitute an ICGEB Alumni body of over 3500 individuals from more than 60 countries around the globe, with an estimated 750 PhD awardees among them.

As we celebrate European Youth Goals, the Arturo Falaschi Fellowship Programme continues to expand in line with the UN 2030 Agenda and the Sustainable Development Goal 4 for Quality Education.

This article was originally published in the Regional Cooperation Magazine, November issue

Maria Juul Diekmann has always had a huge passion for human health. She was always interested in infectious diseases and especially how to prevent them or treat patients that become infected. This is what motivated her to study medical and molecular biology at the University of Roskilde in Denmark and to choose research projects on cancer and bacterial human pathogens as training activities during her Master’s degree. “Many of my colleagues wanted to do these experiences in big Pharma companies but I wanted to see how research works,” she says. This same passion moved her toward the Tick-Borne Flaviviruses network project. “I knew what ticks were and that they can carry some diseases, but I had no idea how harmful tick-borne flaviviruses can be”.

Mainland Denmark is not endemic for Tick-Borne Flaviviruses however Bornholm is considered endemic since the early 1960s. According to the European Centre for Disease Prevention and Control, almost zero TBE cases are reported in the mainland annually. A notable exception is Bornholm island where an incidence of between 4 and 8 cases per 100,000 inhabitants are reported. This is probably why there are very few research groups that study these diseases in the country.

When Maria had to decide on the topic for her Master’s thesis she came into contact with the research field of tick-borne encephalitis virus. “I went to speak with my former professor, Karen Angeliki Krogfelt, at the University of Roskilde and discussed options for a research internship. She has a big network of scientists among her contacts, and I thought she could suggest someone that was doing research that could be interesting for me. Among all the colleagues she mentioned suddenly it was there: a research group in Norway was studying tick-borne encephalitis virus. Tick-Borne Encephalitis virus is endemic in Norway along the coastline with 20-70 cases annually. That was a breakthrough moment for me, I thought: what is this? A virus that through a tick’s bite can enter the brain? That was impressive, and I thought that was what I wanted to study!” But there was a problem: Maria needed to move abroad to Oslo, to the Norway Institute of Health to study what she was interested in. 

“When I was at University, I saw my friends coming back from their Erasmus experiences in Scotland, Spain or elsewhere, and they were so enthusiastic. I saw them so changed and grown, and with so many experiences lived through in just six months. I think I was envious in a kind of way, but I was also scared to do the same. What if I went abroad and then could not find a way to come back to my home country in the future? Or worse, if I went away and discovered that science is not what I want to do in life? These thoughts blocked me in away from taking an Erasmus during my whole study period. But then at some point, the time came to decide on my Master’s thesis internship. My professor told me about this excellent team working on tick-borne encephalitis in Oslo. Well, I thought that it was the right moment to take courage and at least try going abroad.”

Erasmus+ is the most known exchange program for students in the European Union. It is the only one funded directly by the European Commission and under this umbrella students at all stages and from all disciplines can have an educational experience abroad. The year 2022 marks the 35th anniversary of this successful program that over the years has had more than 10 million participants. This is a massive number of people that through the years have experienced studying or volunteering abroad. In 2021 alone there were 640,000 people who took this opportunity. 

In the framework of the TBFVnet project, many student exchanges between the partners had been planned. This is pivotal to exchanging expertise between the different research groups. Who is better trained than graduate and undergraduate students to learn quickly? Maria was in fact meant to take part in an exchange program between the Norway Institute of Health and the International Centre for Genetic Engineering and Biotechnology (ICGEB). “I was supposed to go to Trieste, Italy (where ICGEB headquarters is located) to learn more about the diagnostic tools for tick-borne flaviviruses. But then COVID-19 came and everything stopped”. Maria was just one of the students that were meant to do an exchange period between the different project partners. In line with the vision of the Erasmus program and with the idea that exchanging knowledge is the basis of scientific research, many students had planned to move to different project partners. With COVID-19 all such travel was limited but with the easing of the pandemic, this exchange program is back. Some students will go from Trieste to Oslo and Prague, in the Czech Republic, and others will go from Chisinau, in the Republic of Moldova, to Trieste. These exchange periods will last only a few months but will constitute a breakthrough for each of the students and for their research projects on tick-borne flaviviruses.

In spring 2023 there will also be a regional seminar to gather all project partners, group leaders and students. This will be a great occasion to meet collectively and share knowledge and updates. Moreover, top scientists in the field of flaviviruses will be invited to present a broad and up-to-date view of this evolving research field.

Maria graduated in January this year with a thesis on the tick-borne encephalitis virus spread in Larvik, Norway. This is a suspected endemic area for TBEV in Norway. The research group found out that there are some relations between the climatic changes in the area and the distribution of TBEV. In particular, it seems that a high temperature and low humidity in the environment resulted in a lack of detectable TBEV among the collected ticks. Moreover, she established a method to improve the diagnostic tools in order to distinguish between samples from vaccinated and infected patients. Maria will present the results of her Master’s thesis at the annual TBE conference in Vienna in November 2022 (ISW-TBE 2022). Now she is applying for a PhD to continue her studies next year on TBE and the improvement of Tick-Borne Encephalitis diagnostic tools. Somehow, she has changed her mind about studying abroad. “I definitely recommend doing a period abroad for graduate or undergraduate students. I learned a lot in Oslo, also because I think there are many ways to perform science, many more than the only one you can experience by staying within your own University where you grew up as a student. In the end, I believe that with this experience I have grown, not only as a scientist but also as a person compared to who I was before.”

This article was originally published in the Regional Cooperation Magazine, August issue

On June 21 – the Tick-Borne Flaviviruses network welcomes new partners from the Republic of Moldova. During the online meeting between project partners, the Veterinary Research Institute of the Czech Republic, the Biomedical Research Center of the Slovak Academy of Science, the ICGEB and the Norwegian Institute of Health, presented their expertise in the field of tick-borne flaviviruses research, diagnostics and surveillance. Scientists from the Nicolae Testemitanu State University in Chisinau and the National Agency for Public Health of Moldova were invited to the meeting following their expression of interest in the topics of TBFVnet.

TBFVnet is a network of scientific research institutes across central and eastern Europe with the aim to study and survey tick-borne flaviviruses. The power of both survey and research relies on the collaboration between different institutes in as many countries as possible. One of the most promising objectives of TBFVnet project was to gather new research institutes in the network that could bring new knowledge and expertise and be a new observatory on tick-borne flaviviruses in Europe. This objective was reached in June with the entry of two new partners from the ICGEB Member State Moldova. Moldova is one of the countries in Europe within the endemic region of TBE and other tick-borne flaviviruses. It was actually thanks to the contacts ICGEB has in the country that two institutes, the Nicolae Testemitanu State University in Chisinau and the National Agency for Public Health, got to know about the network.

During the meeting, the TBFVnet partners presented their expertise and research fields to the Moldovan institutes. By their side, Elena Deseatnicova from the Nicolae Testemitanu University and Olga Burduniuc from the Moldovan National Agency for Public Health presented the situation of TBE in the country in terms of cases and affected regions as well as the diagnosis method they currently use. There was a great interest in setting up the diagnosis tool based on the framework of TBFVnet by the Norwegian Institute of Health.

In the near future, there will be exchanges of protocols, and knowledge to improve the research and surveillance of tick-borne flaviviruses in Moldova. With the improvement of the pandemic situation probably some scientists from the two Moldovan institutes will be hosted in one or more of the institutes of TBFVnet.

Beside this good news the TBFVnet have to say goodbye to two of its former members the Chumakov Research Institute from Moscow and the Umea University from Sweden. Our partner from Moscow was forced to leave the project due to the sanctions from Europe to Russia.

A war quickly reaches every aspect of society, and we can certainly say that war is the nemesis of international cooperation – just as science aims to be its champion. We, from the TBFVnet project, were overwhelmed, as was everyone in Europe, by the news coming from Ukraine in recent weeks. Initially, we hoped that our project could be immune to the consequences of this war. We believed science to be immune to war. We thought that since it has no borders, scientific collaboration could continue beyond politics, economic crisis and maybe even war, this was. We were naive and soon this crisis had big and direct consequences for our project.

We expected a lot from our project in the coming year. During the past two years of the pandemic, we struggled to see each other and waited for calmer and safer periods to organise meetings and workshops to share scientific knowledge on tick-borne flaviviruses amongst ourselves and with our peers. We were truly excited by the prospect of being able to do so. One of the objectives of our project is to enlarge our network with the aim to foster the surveillance of tick-borne flaviviruses in many other countries in Europe. In this context we were organising a large meeting in Chisinau, the Republic of Moldova to take place in May 2022. We had no project partner in this country, but some research institutes had expressed an interest in joining the network. This premise led us to believe that a new partner could be the Nicolae Testemitanu State University (USMF). Regrettably, all of this disappeared with the war. The meeting is being rescheduled, hopefully to September. In reality, we cannot be sure when or if it will take place.

However, the most challenging aspect of the TBFVnet project has been that one of our partners, the Chumakov Institute, is based in Moscow. Soon after the war started and Europe decided to shut down all economic support and relationships with Russia, it became clear that this would affect our collaboration. In fact, our Russian partner was forced to leave the project as it could no longer receive payments from the Regional Cooperation Fund. Notwithstanding this setback, the collaboration continues with partners in the Czech Republic, Slovakia and Norway hoping to be able to re-establish connections with our colleagues in Moscow in future, hopefully peaceful, times.

We need to be honest: ticks, as well as the viruses they can carry, are the very least of the problems being faced by the people of Ukraine at this time. Both soldiers and citizens literally fight to survive, and the consequences of a tick bite are not considered a health priority. However, we should also point out that the whole health system in Ukraine is being heavily affected, including the ability to diagnose infections at large, survey infected animals and instruct healthcare providers. It will take years to rebuild all this, while pathogens still circulate across borders. It will be the responsibility of networks such as TBFVnet to reinforce their activities to maintain high attention in the Region and to provide support also towards refugees fleeing war.

One could also say that science and cross border cooperation is not big matters to lose. However, they required years, perhaps even ages, to build. And many many years will be needed when this war comes to an end to rebuild cross border cooperation and scientific collaboration.

When Western countries established sanctions against Russia for its invasion of Ukraine also many international scientific institutions followed along the same line. A notable example is the CERN Council which decided to suspend Russia’s observer status. CERN has made scientific collaboration the core of its mission: it was funded “with the aim of bringing nations and peoples together around science in a spirit of peace”. Clearly, this war affects the fundamental aspect of this international organization. Many Russian scientists have risen up against their own government and its war to stand with Ukraine, the international community and peace.

As the former Director-General of our partner ICGEB, Mauro Giacca, recently wrote for a newspaper in Italy: “It is really difficult to understand whether it is more productive to keep Russian scientists within the international community or to penalise them so that their institutions become vocal in demanding change. Either way, the notion that you can do something in life that has no political implications, including science, remains a myth that again breaks hard against reality.”

Usually, we do not know what scientific research can bring in the future, but oddly, we now risk that nothing could be the answer. Nothing will ever be discovered, explained or developed where there is no collaboration. It will be our responsibility to maintain active all scientific networks of collaboration to minimize this risk and to be able to recover as soon as possible.

This article was originally published in the April issue of the Regional Cooperation Magazine.

In this video, Alessandro Marcello and Tea Carletti leader and research assistant at the Molecular Virology laboratory of project partner ICGEB explains the TBFVnet project.

TBFVnet involves many researchers across Europe that share and try to find new solutions to survey, diagnose and prevent tick-borne diseases caused by flaviviruses.

https://youtu.be/bVck1VOJsgI

Riccardo loves walking in the mountains in spring and summer. Last July he went hiking in Carina in the Friuli Venezia Giulia region in Italy. It was a hot sunny day, so he wore short pants and a short-sleeved shirt. Riccardo took some friends with him to show them the mountains he loves. They had a very long walk amongst the woods and peaks and they rested in the grassland. Once back home, having a shower, Riccardo felt something strange beneath his hand while washing his leg. He saw a small dark freckle on his left calf that wasn’t there before. It would have been difficult to see it without feeling it with his hand. Unfortunately, that was not a piece of dirty dust but a tick that bit him during the walk.

Riccardo felt a bit foolish: “I should have worn longer pants” he thought. However, he was somewhat prepared. He took a fine-tipped tweezer a grabbed the tick firmly. It was difficult to grasp it as close as possible to the skin, but with the help of a magnifying glass, he managed to do so and pulled the tick upward to remove it. Then he finished his shower, applied some antiseptic on the wound and went to sleep. He had a bad night. He dreamt of dangerous diseases spreading up through his leg. The day after he called the doctor who said: “Mark the day on the calendar and call me back if you develop flu-like symptoms within a month. Be more cautious next time, Riccardo!”. The doctor knew this could be a dangerous bite. While the probability of developing severe diseases is low, it is always better to be cautious and monitor the symptoms.

Tick-borne encephalitis (TBE) is a common tick-borne disease, along with Lyme borreliosis. Over the last decade, TBE cases have grown consistently in Europe. According to the European Centre for Disease Control, in 2019 there were 3246 confirmed events of infection. This disease is endemic in many countries of central, north, and eastern Europe where most cases are reported. Unfortunately, Riccardo went hiking in one of the regions where this disease is endemic, i.e. constantly present. While this does not mean that all ticks in that area carry the pathogen, it is a sign of danger, so anyone engaged in outdoor activities should pay higher attention to ticks and tick bites. Tick-borne encephalitis causes flu-like symptoms such as headache, fever, pain, fatigue, and weakness but can have much worse consequences that could lead, in some cases, to death. The disease is caused by a virus, named tick-borne encephalitis virus, that passes into humans from the tick bite and replicates in the body with the risk of causing severe inflammation of the brain, called encephalitis, if not contained efficiently by the immune system. There is no specific treatment for TBE once you are infected but there is an effective vaccine that prevents infection.

Ticks and the diseases they carry can have a great impact both at the human and animal levels. Riccardo and his friends, just as everyone engaged in outdoor recreational activities, including farmers, are at risk in endemic regions. But tick-borne diseases also affect animals and these threats could have great consequences at the economic level, not just for the health system but also for the production sector. Costs derive from the direct loss in the production of meat, milk, eggs and leather that are associated with both simple tick bites and eventual subsequent diseases. Farmers spend great sums of money on therapies when animals become infected and fall ill; in the worst scenario, they could also lose their entire livestock and farm. The annual economic cost of tick-borne diseases has been estimated to be at around 70 million USD, considering only small ruminants.

This is due also to the fact that there are many diseases caused by viruses or bacteria carried by ticks. Tick-borne encephalitis virus is the most known flavivirus, but many others can be harmful to humans and animals. At present only one vaccine is available to protect humans from TBE and much research is needed to develop and find new vaccines for known, and emerging tick-borne diseases. As the current COVID-19 pandemic has shown us, scientists need to share knowledge and information as fast and as openly as possible. When this happens, preventive solutions, treatments and hopefully vaccines can be found quickly. But research is only one part of the solution. The second pillar for successful protection from tick-borne diseases is surveillance. A diffused network of trained surveillance institutes could constantly monitor the presence of known and new pathogens.

The TBFVnet project was launched to address both of these issues. It brings together research institutes across Europe to study and survey tick-borne flaviviruses. The partners are experts in different fields of infectious diseases carried by wild animals and are located in key positions within endemic regions of tick-borne diseases. The partners can study all the different aspects of tick-borne diseases such as the survey of emerging viruses, their biology, and the development of new vaccines as well as new diagnostic tools and antivirals.

International projects like TBFVnet are great examples of how cross border scientific cooperation can help society in the long term. New tools to detect flaviviruses, or effective antivirals, can have a tremendous impact on health. “With the COVID-19 pandemic, we learned too late the importance of quick diagnostics and capillary surveillance for infectious diseases – points out Alessandro Marcello, head of Molecular Virology at the ICGEB – A network of laboratories sharing tools and knowhow should be already in operation on the territory to monitor endemic and emerging diseases to instruct an efficient health response”.

The costs of prevention and research are undoubtedly high. Many scientists and much equipment, as well as transnational infrastructure, are required to face such threats. However, as the COVID-19 pandemic has shown, the cost of prevention represents just 2% of the economic damage caused by an outbreak. Riccardo was lucky, the month passed without him developing any symptoms. But he will remember the dreams he had that night and next time he will be more cautious. He will ask at tourist offices if the region he wants to explore is endemic for tick-borne diseases. Probably he will wear long pants and shirts, use repellents for his clothes, and he will check and tell his friends to check for tick bites at the end of the day. This is what he can do to prevent harmful infections while having a pleasurable, long walk in the countryside. But science can do much more. Research can produce treatments and vaccines to cure sick people and protect the weak.

This article originally appeared in the June issue of the Regional Cooperation Magazine.

Department of Virology at the Norwegian Institute of Public Health (NIPH) has been the National Reference lab for Tick-borne encephalitis virus (TBE virus) since 2008. The reference lab is divided into a routine and a research lab group. The NS1 TBE research group consists of Maria Juul Diekmann (Master student, in collaboration with Roskilde University), Katrine M Paulsen (Chief engineer), Alaka Lamsal (PhD student) and Rose Vikse, Åshild K Andreassen (senior scientists/Professors) as well as external Professor Karen A. Krogfelt (Roskilde University, Denmark).

Our gold standard method for TBE detection and confirmation is serology with the Enzygnost anti-TBE/FSME/ETG (IgG, IgM) kits (Siemens, Marburg, Germany). Due to the discontinued production of these kits, NIPH had to evaluate and validate other commercial methods. The main aim was to find a method with similar sensitivity and specificity as the Enzygnost anti-TBE/FSME/ETG (IgG, IgM) method. In addition, we wanted to find a method that could distinguish between vaccinated and infected TBE cases. The recombinant NS1 protein in the TBF NS1 ELISA assay (developed by ICGEB) is designed to detect flavivirus antibodies with reduced cross-reactivity and high sensitivity and detection of vaccine breakthrough cases. Detection of the difference in the TBE immune response between infected and vaccinated cases is important and we decided to test the non-commercial TBE NS1 ELISA assay together with the commercial ELISA tests available on the market.

Our material consists of 146 random IgM/IgG serum and spinal fluid samples from previously examined cases including quality control samples known to cross-react with the TBE antibodies.

Samples	IgM	IgG
Serum
Positive and borderline	19	20
Negative	17	16
Cross reacting	9	13
Spinal fluid
Positive and borderline	14	16
Negative	11	11
Cross reacting	–	–
Sum	70	76
Total = 146

In addition, we have 12 patient serum samples from a collaborative study with the University of Oslo analysed by the same method (Enzygnost) and confirmed by virus neutralization test (VNT) at the Veterinary Research Institute in the Czech Republic. Some of these sera may be from individuals who are vaccinated by cross-reacting agents i.e. other flaviviruses than TBE.

Preliminary results:

Our master student has performed several initial tests to customize the TBE NS1 ELISA. There has been some trouble in finding the best replacement reagent available since the HRP-linked goat antibodies anti-human IgG from Sigma (A6029-1ml) was not available at the time we started testing. Some strong TBE positive, vaccinated and negative samples were tested with different blocking solutions of 2% BSA and different dilutions of the new secondary antibody HRP-linked goat antibodies anti-human IgG (Fc specific) from Sigma (A0170-1ml). We experienced some high background problems that were solved by a fresh 2% BSA and therefore the new secondary antibody was titrated. In our hands, the dilution suggested from the protocol was too high with the new secondary antibody. New tests will be performed shortly in order to establish the best dilution concentration when using our chosen secondary antibody.

Thanks to this work, we are now able to distinguish between the vaccinated, infected and negative samples and can start to validate the test samples.

Heterogeneity in virus populations and immunosuppression of the host could cause tick-borne encephalitis virus vaccine failures. These are the results of a study recently published in Microorganism by the Karganova group, the TBFVnet partner from Moscow.

The only vaccine currently available to prevent tick-borne diseases is for tick-borne encephalitis virus (TBEV). However, some cases have been reported in which this vaccine failed to induce effective immunity against the disease. The research group led by prof. Karganova, the TBFVnet partner from the Chumakov FSC R&D IBP RAS in Moscow, recently investigated the reasons for this problem. According to their results, TBEV vaccine failures are explained by two factors: virus heterogeneity and immunosuppression of the host.

Vaccine failures occur when people who are completely vaccinated become infected and develop diseases symptoms. Few cases have been reported for TBE vaccinated people. Usually, these are characterized by a mild clinical course, but in some cases, the effects are severe or even lethal. Investigating the causes of these failures is pivotal to enhance vaccine effectiveness and moreover, can give insight into the mechanisms of virus and host interactions.

The study has revealed two main reasons on the basis of vaccine failures: heterogeneity of virus population and host immunosuppression. TBE virus populations are not uniform within a given geographic region and show some structural differences. The more different the virus particles are within a population, the more difficult it will be for the immune system to react to new infections. These differences, in fact, could mislead the immune system and cause failure in protecting the organism. Moreover, if the host immune system is even slightly depressed before vaccination this could impair proper immunity against the virus.

The results of the study could lead to the development of more effective vaccines against TBEV. Moreover, this evidence could help in the future production of effective vaccines for other tick-borne flaviviruses. The research article has been published in Microorganisms in May 2021.

Viruses don’t care about human geographical borders. The coronavirus pandemic made it clear how fast even great distances can be covered by viruses within days. The same is also true for tick-borne diseases and, in particular, for flaviviruses. Tick-borne flaviviruses can move far from their original region carried by wild mammals. Several societal and environmental factors can force deer or other small wild rodents to migrate to more favourable habitats. These animals naturally carry ticks, and along with them, they also carry pathogens including viruses. This makes it clear that borders are only a human need: animals and viruses can cross them easily without leaving traces. Unless they carry pathogens: when this happens, they do leave traces of their passage in human health.

The SARS-CoV-2 virus also showed us the importance of sharing knowledge, and surveillance. Both are pivotal in facing threats to human health. In the tick-borne flaviviruses scenario, a constant survey of ticks, pathogens and animals is crucial to monitor how diseases spread in new areas. These activities are much more effective when performed within a framework of international cooperation. The TBFVnet project was established to foster scientific collaboration among research institutes across Europe to study and survey tick-borne flaviviruses. Each partner institute excels in a different field of study of flavivirus biology and distinct expertise can make the difference.

The TBFVnet partner at the Veterinary Research Institute of the Czech Republic is an expert in the pathogenesis of the Tick-Borne Encephalitis virus and the development and testing of vaccines and antivirals. In Russia, the Chumakov FSC for Research and Development of Immune-and-Biological Products focuses on the surveillance of Eastern strains of TBFV as well as on antivirals and vaccines. Scientists at the Biomedical Research Center of the Slovak Academy of Science study the ecology of new flaviviruses. The Norwegian Institute of Public Health monitors TBFV locally and develops molecular and serological diagnostic tools including sequencing of viral genomes. The partner from Umeå University, Sweden, is an expert in the virus-host interactions and neurological aspects of TBFV diseases. Finally, the ICGEB in Italy develops and studies virus-host cell interactions, novel diagnostic and surveillance tools, and antivirals. In addition, the ICGEB is actively involved in expanding this network to neighbouring countries.

Within TBFVnet, partner institutes do their best in sharing protocols and practices to study and survey tick-borne flaviviruses. One of the most successful activities is the regular online meetings to present, share and discuss laboratory procedures to produce antibodies, isolate viruses, and other techniques to study tick-borne flaviviruses. The efforts of the TBFVnet project go beyond research. The established network is willing to do its part to enhance the benefit for the whole society with a clear vision of the role of science in the knowledge society and economy. Collectively, the partners are able to capitalise on scientific discoveries and basic and applied research. Society can benefit from these efforts thanks to the transfer of this knowledge from research to industry and effective medical applications.

This article originally appeared in the June issue of the Regional Cooperation Magazine.

You may have heard about the book Spillover by David Quammen. In a nutshell, this fascinating book describes how diseases can emerge from animals and spread to humans. An entire chapter is dedicated to the story of Lyme disease, a disease caused by a bacterium. Lyme disease affects approximately 65,000 people in Europe and can give rise to long term consequences including fatigue, pain and memory problems. The name comes from the city of Lyme in the US where the disease was first characterized.

There may be no apparent reason to talk about Lyme disease on a website dedicated to flaviviruses. Except that there is one: ticks. Borrelia burgdorferi, the bacterium causing Lyme disease, infects humans trough ticks. A tick may bite a human in search of a fresh meal of blood but if it is infected, while feeding it leaves the bacterium to pass into human blood, where it stays giving rise to the first symptom: a red rash at the site of infection.

Unfortunately, Lyme disease is not the only tick-borne disease. Like Borrelia burgdorferi a number of other bacteria and viruses can be carried by ticks and some are severe human pathogens. One of these is tick-borne encephalitis virus, TBEV, that causes the tick-borne encephalitis. It belongs to flaviviruses a family of viruses that takes its name from its most notorious member: the virus that causes yellow fever. However, many flaviviruses find their vector in ticks between hosts and thereby they also reach humans. These viruses are the research subject of the TBFVnet project.

TBFVnet stands for Tick-borne flavivirus network. Researchers from six different institutes across as many countries in Europe work together to study and survey flaviviruses carried by ticks. Beside TBEV, the project focuses its efforts on three other tick-borne flaviviruses that affect the human population: the Powassan virus (POWV), the Louping-ill virus (LIV) and the Omsk hemorrhagic fever virus (OHFV). In 2019 in Europe there were 3246 confirmed cases of TBE alone (with an incidence of seven per million of habitants) marking an increase over the past years as reported by the European Centre for Disease Prevention and Control (ECDC) dedicated page. At the moment a vaccine is available only for TBE, while for other tick flaviviruses there is no vaccine or specific treatment. The TBFV network aims to fill this gap sharing expertise to understand their biology, develop treatments and diagnostic tools.

The problem with tick-borne diseases is the tick itself. They are quite difficult to see, and they can bite anyone engaged in outdoor activities such as hiking or farming. Unfortunately, it is difficult to sense a tick bite, most of the time you could notice the tick on your skin just when you are back home. However, and here comes the first good news, even if ticks can be anywhere outdoors, the pathogens are not. Viruses carried by ticks have their own geographical distribution that is monitored and visible through local authorities and on the ECDC website. Surveillance data are important to understand the distribution of a pathogen and to establish risk areas and implement countermeasures. However, monitoring capacity is unevenly distributed in different countries and TBFVnet aim at filling this gap.

More good news follows at the end of this story. First, in a given area just a small fraction of ticks carry the pathogen. So, the probability of being infected is low. The second is even more important because it is something each of us can do: protect ourselves. This is very easy, because the moment we are aware of this threat we can do something. On the EDCD website there is a section listing personal actions to protect ourselves. For example, we can check whether there are any known tick-borne diseases in the area we are planning our outdoors activities. Even more easily, we can wear suitable clothing: long trousers and long-sleeved shirts can easily protect our skin from ticks. For those who want to feel even safer, there are also insect repellants that can be applied on boots or clothes to raise the level of protection.

In Norway they say, “There is no bad weather, only inappropriate clothing”. At the end of this story, we can borrow this saying and remember that there will be no detrimental tick bites if we prepare.

The TBFVnet project represents a network of laboratories that aim to study and survey tick-borne flaviviruses, involving six institutes across Central Europe, at the heart of the endemic region. In December 2020 the tbfvnet.eu website has been launched.

The TBFVnet project is funded by Iceland, Liechtenstein and Norway through the EEA and Norway Grants Fund for Regional Cooperation. The Project aims to build an international network of laboratories that collect and share expertise, tools and protocols to investigate the biology and the pathogenesis of Tick-Borne flaviviruses (TBFV). The project operates through six institutes across Central Europe, at the heart of the endemic region: the Czech Republic, Italy, Norway, Russia, the Slovak Republic, and Sweden. In December 2020, the dedicated TBFVnet website was launched: tbfvneticgeb.local/.

TBFVs include a large number of human and animal pathogens causing severe disease. The project focuses on Tick-Borne encephalitis virus (TBEV), Powassan virus (POWV), Luping-ill virus (LIV) and Omsk hemorrhagic fever virus (OHFV). At present no antivirals or treatments for TBFV infections are available and a prophylactic vaccine is only available for Tick-Borne Encephalitis virus (TBEV). 

TBFVnet aims to develop novel diagnostic tools and antivirals against tick-borne flavivirus infections. Efforts will also aim to raise awareness on tick-borne diseases, and involve research centres from other countries to join TBFVnet. 

The Project was recently announced by ICGEB, which heads the Communications work package, at its annual Board of Governors session, which brings together representatives from 65 Member States worldwide, and at its Council of Scientific Advisers, which also includes leading scientists and Nobel laureates active in TBFV endemic countries, including the USA.

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A paper published by the Molecular Virology Group, ICGEB Trieste, Italy, presents a new strategy to diagnose flaviviruses infection in patient sera. This work is pivotal for the TBFVnet project as starting point to develop new assays for tick-borne flaviviruses.

The Molecular Virology Group is led by Alessandro Marcello at the partner institution ICGEB. The Group has developed a new assay to detect flaviviruses antibodies in human sera, from which it will be possible to study the development of new assays for other tick-borne flaviviruses.

Diagnosis of flavivirus infection through virus isolation, detection of viral RNA and viral antigens are highly specific but limited to the short time window of the viremic phase of the infection (2-3 weeks). Serological assays provide a longer time-window for testing. IgM antibodies can be detected in most patients by day 3-5 after the onset of symptoms and remain for several months following infection. IgG antibodies are initially detectable at low titers but are stable for longer time periods.

Present serological assays for flaviviruses are mostly based on structural antigens, while the proposed assay suggests the non-structural protein 1 (NS1) as a novel target. This protein is secreted in high amounts during early stages of viral replication and causes an immediate immune response with anti-NS1 IgM (2-3 days after infection). The authors describe a strategy to produce and purify high quality, non-structural protein 1 (NS1) to use in an ELISA test to detect the presence of flavivirus antibodies in patients’ sera. The results clearly show that NS1 is a valid antigen with higher sensitivity as compared to previously available commercial assays. Furthermore, this assay is also able to detect infection in patients with vaccination breakthrough TBE, particularly in older individuals.

The expertise developed at ICGEB during this research will be shared with the partners of TBFVnet through in-site and on-line training. Combined efforts will boost the development of similar assays to help fast monitoring and surveillance of tick-borne flaviviruses.

The article by Marcello and colleagues has been published in PLOS Neglected Tropical Diseases.

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The Laboratory of Arbovirology, Veterinary Research Institute, Czech Republic, recently developed a new tool that can boost the search for new antiviral agents and detect neutralising antibodies for TBEV. The Group has engineered TBEV with mCherry fluorescent protein to be used in in vitro screening.

The Laboratory of Arbovirology led by Daniel Ruzek TBFVnet partner recently published a paper with great impact on the TBFVnet project. The research presents a new tool that can identify novel antiviral agents and detect neutralising antibodies for tick-borne encephalitis virus.

Tick-borne encephalitis virus (TBEV) is a flavivirus widely spread in large regions of Europe and Asia. This virus can cause a neurological disease known as tick-borne encephalitis (TBE) in humans. It is estimated that 12000 cases of TBE occur annually. Even though the mortality of TBE in Europe is estimated to be less than 1%, many patients show long-term or even permanent consequences known as postencephalitic syndrome. TBE is preventable with vaccines but in several endemic countries, vaccination coverage remains low. Moreover, there are no approved antiviral drugs to treat patients, although several candidate antivirals effective against TBEV have been identified. At present the study of this virus requires laborious secondary approaches to detect the presence of the virus in infected cells and/or in animal models of infection.

The research published by Ruzek and colleagues presents a new strategy to overcome this problem. They have designed and constructed a recombinant protein to be used as a reporter in antiviral screening for TBEV. In particular, they engineered TBEV with a monomeric red fluorescent protein the mCherry protein. This reporter gene is extremely useful in monitoring in vitro infection and its expression remained relatively stable during several passages in cell culture. Thus, the mCherry-TBEV is a unique opportunity for high-throughput screening of antiviral compounds, and the detection of neutralising antibody in cell culture systems.

These findings are extremely important as TBEV is transmitted by ticks of the Ixodes genera: one of the main targets of the TBFVnet project. This study in the framework of such collaborations can accelerate the development of new treatments and diagnostic tools. 

The research has been published in Antiviral Research.