Professor Krystyna Bieńkowska-Szewczyk talks about Poland's most modern BSL3+ laboratory established at the Intercollegiate Faculty of Biotechnology UG and MUG and discusses working with viruses in an interview with Julia Bereszczyńska.

Julia Bereszczynska: - BSL3+ Laboratory. This acronym sounds quite mysterious to someone who does not deal with virus work on a daily basis. What does it stand for?

Prof. Krystyna Bieńkowska-Szewczyk: - The acronym comes from English and stands for Biosafety Level. It is about safety when working with organisms that are a potential threat. There is a scale from BSL1 to BSL4. BSL1 means that it works with minimal safeguards. BSL2 works with pathogens that require some protection but no special isolation, i.e. with viruses such as our model animal herpesvirus, which is very similar to the human herpes virus but is not pathogenic to humans. In contrast, when we proceed to work with a live virus that is potentially pathogenic to humans, the laboratory must be very carefully isolated. There are several regulations and specific rules for the construction of such a laboratory. We in our department have so far had a BSL2-class unit. Such a laboratory is separated from all others by a system of several doors, and only authorised and appropriately dressed people can enter. Viruses are worked with on-site in special chambers called laminar chambers. These are characterised by a layered, or laminar, airflow that passes through filters. Therefore, inside such a chamber, the workplace is sterile, and the person working there is separated by glass and completely safe. We found ourselves in a situation where there was a coronavirus pandemic, and coronavirus, as we all know, spreads by air, by inhalation. There were two aspects that were an obstacle to working with this virus. Firstly, we lacked the safeguards to work with such airborne pathogens. So, we had to redesign part of our laboratory and upgrade its security status. Secondly, the virus itself caused great fear. At the moment, the second aspect is no longer so important because everyone has been vaccinated, many people have been ill frequently, and the approach to working with SARS-CoV2 is more rational. Adapting the laboratory was a major undertaking, and in the end, we achieved a very high level of security, described as BSL3+.

- Is that the highest level?

- The highest is four, and no such laboratory exists in Poland. Under such conditions, the most dangerous viruses, such as Ebola, should be studied. A laboratory of this type is a completely closed unit. This means not only limited access to it but also working in special hermetic suits with individual air supply. In such a unit, everything is sterilised, including the water from the toilets. As far as I know, there is no fourth-level laboratory in Poland. Perhaps the army has such a place? But I don't think so. I think this is a big mistake because if some completely unknown virus were to appear, it would be good for such a laboratory to exist after all.

- What viruses will scientists be able to deal with here?

- It will primarily be coronavirus. We will also be working with tick-borne encephalitis virus. This is a virus that is quite a dangerous human pathogen. We have been working with it in a very limited way because it is all about live virus multiplication. It is similar to the Zika virus or certain strains of influenza - they also require this special protection. The next virus is the hepatitis C virus. These are viruses with which work depends on the profile of actions. Our laboratory is not clinical; it is not for diagnostic purposes. It is a research laboratory. We work with live viruses, so it is necessary to culture different types of cells because viruses can only be multiplied in live cells.

- Will only highly trained staff have access to the laboratory? Will students also be able to use this state-of-the-art infrastructure?

- I can imagine that a student preparing a master's thesis would be trained in this respect; however, the BSL3 laboratory is a place where many employees cannot work simultaneously. There can only be a few because of the air exchange issue. So, we are talking about a relatively limited group of people.

- I was just going to ask about the safeguards preventing viruses from getting out of the laboratory.

- You could say that we have built a second laboratory inside the existing one. It has steel walls and access is through two man locks, so the air exchange is controlled. The principle of building laboratories of this kind is that a negative pressure is present in them - the pressure is lower than in the environment. So, if all these barriers were to break, the air would be absorbed inside instead of escaping outside. The walls, by the way, are also double, and we monitor the pressure in real time. We have a camera that shows what is happening inside. Because of the amount of air, a maximum of five people can stay inside. They have to work in special outfits at all times, and certainly, no untrained person can enter.

- I would guess that water is also needed for scientific research. What about possible contaminated wastewater?

- This has been a difficult task for us. The laboratory is in a building that already exists. In addition, it is located on the third floor. We first thought of breaking through to the sewage treatment plant we have in the basement, which meant interfering with each floor's structure. The problem was solved so that a small sewage treatment plant was set up on the level where the laboratory is. It has a certain capacity and its own monitoring system, so we also managed to secure this issue.

- Despite the safeguards, is it possible for a laboratory employee to carry viruses outside? What then?

- Probably, if they wanted to do it knowingly and maliciously, they would have the possibility, but unknowingly, it is not possible. We will be working with viruses that are altered in some way - they are mostly less dangerous than these wild strains. On the other hand, the regulations say that what is genetically modified cannot be allowed into the environment in any way. This is the primary motivation for what is tested in the lab to be kept in the lab. We have all the equipment on-site - incubators, chambers, refrigerators, and freezers. It's a closed system.

- Are there many scientists who would want to work in a laboratory where research is conducted on such dangerous pathogens? Aren't people afraid of getting infected with something, for example? I remember situations at the beginning of the pandemic when medics stayed overnight in hospitals to avoid going home and endangering children, families... I wonder if scientists would be willing to work in such a place.

- I think any of us would be able to work safely even in lesser security with such a virus at the moment. With our level of safeguards, equipment, and laboratory experience of staff, we have no such concerns, and it is a fascinating area of science. In addition, we have several important projects funded by the National Science Centre that we cannot carry out without multiplying the live virus in specific cells. This is why we have waited so impatiently to meet all the requirements for the operation of such a laboratory. I would also like to mention that the construction tender was won by a contractor from Germany. In Poland, we could not find a suitable contractor for this task due to the fact that there are few such laboratories.

- And is it really the most modern laboratory in Poland?

- It is difficult for me to say anything about other laboratories. However, in a country like Poland, where we have 38 million people, there should be more than a dozen similar facilities, and there has not been such a place in northern Poland so far. There is certainly a fully functional laboratory in Krakow, a laboratory in Warsaw, and perhaps some military ones. For me, a virological laboratory is one in which you can not only take a sample from a patient and check what is in it but also take the virus, multiply it, and conduct experiments with it. Here are people who have such skills.

- Will there be research on any specific viruses in the near future?

- Yes, of course. This will be work on the SARS-CoV2 virus. What interests me is how this virus can get from cell to cell. We have such a project funded by the NSC programme. We know for sure that this virus is able to enter very different cells. We also know that the virus can infect various organs, not just the respiratory organs, as people who have had intestinal symptoms, for example, have found out. Neurological symptoms, on the other hand, are the most misunderstood. Our task as researchers is to understand how the virus gets into cells that don't have the right receptors because, surprisingly, it can enter cells that it theoretically has no right to enter. This is what we are interested in. Dr Lipinska, from my team, has been interested for years in how a virus affects our immunity - because viruses often modulate this immunity, trying to weaken it, for example. Why? To be able to survive in our body. And coronavirus undoubtedly manipulates our immune system. Dr Lipinska wants to join this strand of research, which will perhaps find the answer to which specific proteins cause the immune system to weaken during infection. So there are several projects, and certainly, some need to be conducted in a specialised laboratory.

- Since we are talking about coronavirus, I will ask about vaccines. In your opinion, should we still vaccinate?

- I was vaccinated previously, and, of course, I think we should continue to vaccinate. I was very keen to be vaccinated with the latest version of the vaccine, which has just been done. There is a need for new vaccines. This research that we are doing is necessary because I think differentiated vaccines should come onto the market. Nowadays, there are a lot of vaccines based on live viruses, which are so damaged that they do not cause the disease but give a complete set of immunity. I hope that in the future, such vaccines based on highly altered viruses will come onto the market. Here, the scope for action is very large. The mRNA vaccines are a fantastic invention, but we are somewhat dependent on the pharmaceutical companies and what they want to produce for us. It has to be profitable for them, too. Giving people billions of vaccines has also generated billions in profits for these companies. This is, of course, nothing wrong; these vaccines have had significant effects. I have no doubt that without them, dozens more millions of people would have died from coronavirus. In 2020/2021, we were attacked by the Delta variant; we did experiments recently showing how this variant was able to penetrate cells. It showed incredible activity and intensity of cell penetration compared to earlier variants and the later Omicron one. Admittedly, Omicron also penetrates very easily, but not so easily into the cells of the lungs, but rather into those layers that line our nose or throat. If vaccines had not appeared in that first period, I think the mortality rate would have been much higher because people were getting severe pneumonia.

- However, despite everything, some people did not want to be vaccinated.

- It is with great sadness that I think of several young people who have died because of this. The case of an acquaintance comes to mind: a man in his thirties who strenuously refused to be vaccinated and died from COVID-19. I don't know why exactly he went through the infection so severely, but at the time, these infections could be like that. The severity of the symptoms depends on many things, including how someone becomes infected. If, for example, a heavily infected patient sneezes right in someone's face, the dose of the virus can be so huge that the infection will develop very rapidly, especially if the other person's immune system is temporarily weakened, for example, due to fatigue.

- Returning to the lab, I wanted to ask about the technicalities. What was the cost of setting up this lab? Did it require getting a lot of funding?

- I wanted to get this money since the beginning of the pandemic. Unfortunately, we obtained it far too late. Well, maybe not far, but too late - because we could have done more during the pandemic. We got a grant of four million zloty from the Ministry of Science and Higher Education. Then it turned out not to be enough, and we got an additional one and a half million. This gives us a total of five and a half million zloty. Initially, the high costs were generated primarily by the whole expertise and making a detailed plan. Then, of course, there was the execution itself. We also had to duplicate some of our equipment, such as the special microscope, because it has to be on-site as we can't move samples. We have already made such purchases from university financial sources or grants. It was undoubtedly a costly investment, but I think it's an excellent investment for the future because it opens up the possibility for research that was impossible before, new collaborations, maybe also taking on projects that are partly commercial, but above all, helping people. During the pandemic, we had a lot of requests to see if we could test, for example, virucidal agents or some equipment, and we were not really in a position to accept such requests. So, we did experiments pro bono at the time, using surrogate viruses. The establishment of such a laboratory opens a great many doors. It will allow us to expand the scale of projects in the future. We will be able to apply for grants for projects that require work in such a place. This laboratory opens the door to a completely different world for us.

- I already know how much it all cost. And how long did it take?

- It took a long time because we got this money almost two years ago, and it took a year to build it. Believe me, I put a huge chunk of my life into it. Going to the ministry, convincing and persuading people, writing the application, correcting it according to the requirements from the Ministry... We had dozens of meetings here. It is true that I was the head of this project, but the person who practically supervised it was dr Michał Rychłowski, a scientist with great technical talents. He was the one who coordinated the work of the large team. He is a bit of a pedant, which is why he didn't let a single imperfection pass by, and thanks to him, everything is so perfectly fine-tuned.

- So when is the official opening?

- I think we will manage to have an official opening, to which we will also invite the media.

- Because this is a huge success, Professor.

- It's a huge success. My husband and I were once post-doctoral fellows in the United States and worked with human viruses. I worked with the polio virus, and my husband worked with the influenza virus. When we came to Poland in the late 1980s, it was very poor. There was no chance to work with such human viruses because we didn't have such a laboratory. To multiply the virus I wanted to work with, I used to travel all the way to Puławy because we had started a collaboration with the State Veterinary Institute. It was really far away. First, we brought in one laminar chamber and one piece of equipment, then a second, then a third... I went to conventions, saw what was important, and we gradually got money for things that now seem obvious to people, but times were tough initially. At the moment, we have very high standards for this laboratory. I have to tell you that it is costly. Virology is the kind of science where a person first has to learn biology, cell biology, biochemistry, and various other things, and only then virology. It's a very expensive field. Most of the stuff we use is disposable; everything has to be sterile, after all. Building it is one thing, but it takes several grants to maintain such a laboratory. I think I will leave a very good legacy here. For many years, I have kept my team only by obtaining European grants, which is not a simple matter either. We have done, I believe, a great deal for our university. Maybe thanks to actions like the construction of this modern laboratory, young people will stop leaving. They keep leaving because salaries are very low, and a scientist in our field doesn't have time to work elsewhere and make extra money - laboratory work is arduous and time-consuming.

- Let's assume that scientists in our lab have made a breakthrough discovery. What then? What is the procedure?

- In our field, research results are published in various international journals; we practically do not publish in Polish journals - we aim to publish internationally. It isn't easy to place your results in some journals, and the better the results, the further we go. A good journal is characterised by the scores and indexes, and the higher one publishes, the better the chance to get good money from Polish institutions. The National Science Centre has been very impoverished by the last government. The budget is limited; there were no increases to take inflation into account, and there was no increase in the budget. At the moment, the chance of success is below 10 per cent. In any case, publication is our primary measure of the quality of work. Sometimes, there is also a patent. Then, our aim is for this patent to be not only Polish but also international. In this case, the patent applicant bears the costs, but the example of vaccines has shown that this can turn into huge profits. Mrs Katalin Karikó, awarded the Nobel Prize, fought hard for many years to get the funding, because nobody believed that mRNA modified in this way would be an excellent vaccine. And she proved it. Scientific support is still insufficient, especially for a field like ours, which requires substantial laboratory facilities and execution costs. But at least we can have international cooperation; we can go to conferences, buy reagents from foreign companies if necessary...

- And does the European Union give a lot of support?

- Yes, but there is great interest in the competitions that are announced by the European Commission. These are mostly multi-partner projects. We were, for example, in consortia working on a vaccine for hepatitis C, with a total budget of about 12 million euros. For us, there was, let's say, a few hundred thousand out of this pot, and our Ministry was still subsidising us at the time because this was the policy in order to further strengthen this project. This project was carried out by ten teams, including large pharmaceutical companies.

- Finally, Professor, how did you manage to reconcile your work as a scientist with your family life? Was this difficult for you?

- I am fortunate that my husband works in the same field. We have always worked together and have always supported each other. In the professional area, this support was such that sometimes, only one of us had grants, and we could support each other in running the laboratory, which was very helpful. On the other hand, there were also difficult moments. My son was born in the U.S.; there was no family on site, and there was no maternity leave. After giving birth, I went back to work after six weeks, and only because I had a good boss who let me stay at home for a few weeks. After that, we hired different nannies. Sometimes, it was a difficult task because they were, after all, strangers. I had to leave my child with a person I was seeing for the first time in my life and go to work, despite the well-known fact of how you are connected to your child during this period. When I came to Poland, my child was one year old, and for the first two years, I worked only on teaching and not research; I spent a lot of time at home. My mother also helped me. I know it isn't easy without any support, but we had a unique situation where my husband and I always worked together, which was a big help.

- Professor, thank you very much for the inspiring talk.

The official opening of the BSL3+ laboratory will take place on January 22, 2024, at 10 a.m.