Drs. Jonathan Yewdell, Ivan Kosik, and Alberto López-Muñoz Oral History

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Dr. Jonathan Yewdell, Dr. Ivan Kosik, and Dr. Alberto D. López-Muñoz

Behind the Mask

May 13, 2021

Barr: Good afternoon. Today is May 13, 2021. My name is Gabrielle Barr. I'm the archivist with the Oﬃce of NIH History and Stetten Museum, and today I have the pleasure of speaking with Dr. Jonathan Yewdell who is the Chief of the Cellular Biology Section with the National Institute of Allergy and Infectious Diseases (NIAID), Dr. Ivan Kosik who is a staff scientist in Dr. Yewdell’s lab, and Dr. Alberto López-Muñoz who is a post-doctoral fellow who also works with Dr. Yewdell and Dr. Kosik. Today, they are going to speak about some of their Covid research and experiences. Thank you very much for being with me.

Kosik: Thank you for having us.

Barr: Absolutely.

López-Muñoz: Thank you for having us.

Barr: Will you please speak about how your lab participated in the study that looked at SARS-CoV-2 sero-assay optimization and performance in a population with a high background reactivity in Mali?

Yewdell: I think Ivan you should address that.

Kosik: Yeah, okay I can address that. So ﬁrst of all I'm going to say I have, for a long time now, been a virologist interested in antibodies and viruses, and I'm studying the inﬂuenza virus and now also SARS-CoV-2. I'm looking into how antibodies interfere with the virus, how they affect the evolution of the virus, and what can antibodies say about the architecture of the virus, structure of the virus, and other cool and interesting things. So maybe in late November or December 2020, we got contacted by Dr. John Woodford with requests of “If you would be interested in this study.” They've seen some high background signal in a data set of serum samples from Mali, and they were wondering if we could look at these samples from a different angle, and at the time we had the assay in place for neutralizing the pseudotype of the virus, which would have on the spike on the surface from SARS, but it being a BSL-2 virus. So we of course said yes, and that's how we got involved. They provided us with the set of human sera. They actually saw a very high signal by ELISA [enzyme-linked immunoassay], and we ran those same samples in our assay, and we didn't see any neutralization.

Yewdell: These were from non-infected individuals. There were two aspects: one is why do they have a cross reaction, and two, more pragmatically how can you run screening tests if the population who isn't infected shows up positive?

Barr: That’s very interesting. Were you surprised at all?

Kosik: To be honest with you, no. I was not as much surprised of—this would be the other way. So ELISA, the assay used for screening of the serial positivity, while being used widely and for many decades, people don't pay too much attention to details. It's a tricky assay, and you don't really know what is happening to the antigen on the plate or what areas or surfaces of the protein are exposed. I was not surprised that there's possibility of having a high signal for that while not having any neutralization functional effect with the very same samples in the context of the of the pseudotype neutralization assay. I will be very surprised the other way.

Barr: Okay. Why is ELISA such a tricky assay to work with?

Yewdell: Good question. It's just what Ivan said. I mean there are a lot of assumptions about it that have never really been checked out, like it's not really completely clear why proteins stick to the plastic the way they do, and then once they're stuck the question is, well, how similar are they to the actual proteins that are interacting with the immune system in a living body? Those things are not terribly well established. Ivan has experience, and I have 40 years of experience with this kind of assay. I came from a lab that made the ﬁrst monoclonal viruses to antibodies. It was a lab in the Wistar Institute in Philadelphia. My mentor was a guy named Walter Gerhard. Walter made the ﬁrst hybrid almost to viruses, and so I have a lot of experience with working with monoclonals against inﬂuenza virus. You know, every virus has its own tricks and secrets, but there are a lot of commonalities. Between Ivan and me, we've had a lot of experience with these types of assays, and we know you have to be careful. With that being said, I think what they were seeing is real. It wasn't really an artifact. There is cross-reactivity in serum antibodies in people who are exposed to seasonal coronaviruses as we all are. As we'll get to with Alberto, the fact that an antibody doesn't neutralize the virus in vitro or even in vivo doesn't mean the antibody is not functional as part of the immune response, and one of the messages from inﬂuenza in the last ﬁve or ten years is that there are lots of antibodies that people had not known about or ignored that in the end may prove very important in providing protection against disease.

That's one of the things with SARS-CoV-2 as well is that there's protection against infection, which is called sterilizing immunity, which is important, but if—in a perfect world that's what you would only go for—but in the real world what you're really interested is protection against disease and protection against transmission. There the immune system doesn't have to be perfect, and like we know with infections, most people who are infected, they have a relatively lower level of antibody than those who are vaccinated, particularly with the mRNA vaccines. Those are proving to be pretty much insuﬃcient in many cases to preventing reinfection, but very eﬃcient in preventing serious disease. The vaccines, which is kind of miraculous actually compared to the how people were thinking they would work, the mRNA vaccines certainly induce a better acute antibody response than the viral infections on average. We'll see about the duration, but the odds are they're going to be more durable as well. So these mRNA vaccines are just astonishingly good, and one of the silver linings in this cloud of Covid is that this is pointing the way to making better vaccines for everything basically, including inﬂuenza. Influenza is famously diﬃcult to vaccinate against and most people are infected many, many times in their life with the inﬂuenza virus, and they're vaccinated many, many times, and yet they still get infected. It’s possible the mRNA vaccines will make a real dent in that problem as well.

Kosik: It's not like they've been there, you know, just a year. The mRNA platform is out there for 10 years—2013. There are reviews about the mRNA vaccines. It's just the procedure to making them, manufacturing them is state of the art of science. It's just unbelievable. I mean from a scientiﬁc standpoint, now there is a requirement that we really need to have something really, really good. We’re lucky.

Yewdell: Yeah. So Covid is going to improve vaccination against everything basically.

Barr: That's an interesting point. That's a really interesting point. Ivan how long did it take you to screen those samples with this study, and did you encounter any hurdles or was it sort of routine like other work that you've done?

Kosik: So there's never routine. This is what's amazing about this hobby. I'm not going to even call it a job because it's not a job for me, but the point being, at that time, we had the assay well established, and at the time I ran hundreds of samples already for four different collaborations or our monoclonals we were in the process of making. I got a set of more than 100 samples, and it was done next week basically. I just ampliﬁed the cells. Another post mate looking in a lab of Jaro Holly. You are able to have you know enough, and I run it in three days. With this assay, I want to emphasize that as compared to other assays, the way we are doing this—we have a multiple dilutions of the serum which provides us with the full biology, the neutralization curve [Kosik gestures to represent a concave curve], the measurements of how eﬃcient the neutralization is at given concentration of antibodies. We are able to run 10,000 samples a week easily that would be required.

Barr: It's very impressive. Can you please talk a little bit about how the study was conducted in terms of methodology?

Kosik: Of course. The way this is done, as Jon mentioned earlier, I'll be using the BSL-2 platform-safe virus to work with, and it's a Vesicular Stomatitis Virus, which is genetically modiﬁed so it doesn't express its own cell-attaching protein.

Barr: Did you have to create this or…?

Kosik: No. This is commercially available. What you need to provide, you will take this non-replicated virus, and you combine it with the cell line, which expresses on its surface any viral glycoprotein. It could be inﬂuenza, but in this case it was the SARS spike protein, and when you do that the virus will package. It will infect the cells the ﬁrst round, and it will just start budding after a couple hours. And it will pack the SARS glycoprotein onto its surface, and now you have a stock of the virus, which is not replicative, but uses the spike from the SARS to get inside the susceptible cell line. On top of it, it also expresses green ﬂuorescent protein. When the cells get infected, it will light up green. Now you have this system at which you actually can have the virus, when it impacts the cell, the cells are turning green. If you add the antibody at different dilutions, you will neutralize a fraction of the overall amount of particles you have in the soup, and then less and less cells are infected up to 100% neutralization. Now you take that sample after overnight, which is also a beneﬁt of this assay as compared to other platforms, which take even up to four days. With this assay, you combine antibodies and the virus and the cells in the evening. The next morning, you come into the lab, and you can start measuring. You put the cells into the machine or instrument (ﬂow cytometer), and it will actually collect the positive events and negative events. You can then calculate the fraction of infected cells or eﬃciency of neutralization for a given serum.

Barr: Well that's really interesting.

Yewdell: Yeah, the ﬂow cytometer is one of the great inventions in medical technology. It should have been a Nobel prize for the guys who developed it, but the machines now they can do up to a hundred thousand events per second. You can get the best data you've ever seen in your life with a ﬂow cytometer. Nothing is close. The precision is perfect; the linearity is perfect. In other words, making 10 times as much protein, the signal is 10 times more. There's no other assay that we've ever done that's close to ﬂow-based assays, and a lot of the work in my lab uses a ﬂow cytometer.

Kosik: I don't know a single scientist in Jon's lab who wouldn't fall in love with the flow cytometry.

Yewdell: It's an amazing machine.

Kosik: It's like, you know, I never did that before until I joined John's lab, and now I'm doing it every other day.

Yewdell: Our institute is particularly rich in flow cytometers. Almost every lab has their own, but on top of that we have this fantastic facility for doing really complicated flow things like—originally people called it the cell sorter, and that's how it became ﬁrst famous and it's fantastic for that. It'll take any population of cells and split them into however many populations you can show by the various colors that you use. But on top of that, it's just an amazing analytical machine and NIAID [National Institute of Allergies and Infectious Diseases] has got as much experience as anywhere in the world in this technology, and the VRC [Vaccine Research Center] particularly. There’s a guy there named Mario Roederer who is one of the world's leaders in instrumentation, but also, he was the person who was part of the team that wrote this software that we all use called “Flow Joe”. The NIH is an amazing place for ﬂow cytometry. And we're even using it now; we didn't develop this. You can even use it to measure the ﬂuorescence of individual viral particles. It's incredible, right? So you can actually measure antibody binding to individual, what we call, virions—the individual [virus] particles.

Kosik: Well, not just that, but you can even do the kinetics. You can basically mix the virus with the ﬂuorescently labeled antibody: time zero. You put it onto the instrument, and you can see the increase of the signal as more and more molecular binding, which gives me the goosebumps.

Yewdell: [Laughter] Which in Slovak is called?

Kosik: In Slovak, it is called zimomriavky. [Laughter]

Yewdell: Write that in your transcript.

Barr: Yes. Deﬁnitely true. Dr. López-Muñoz, can you speak a little bit about some of the Covid work that you've been doing these past couple months?

López-Muñoz: Absolutely, it will be my pleasure. Well ﬁrst of all, I would like to brieﬂy introduce myself. I would consider myself a molecular virologist. I am passionate about understanding how viruses are able to mimic and modulate the host immune system. Originally, I joined Jon's lab in order to study or investigate antigen presentation in the context of the inﬂuenza virus infection, but when the pandemic hit the world, we tried to rethink how could I use my expertise in order to address this new emerging virus. We've been trying to investigate non-canonical roles of SARS-CoV-2 major proteins in terms of this immunomodulation or immunoevasion potential role that they could be playing. The idea behind it is that it's well-known and described that there are viruses, usually with large genomes such as a Herpes virus and pox viruses, which encode a wide repertoire of proteins which are particularly dedicated, or they are particularly focused, to target the human immune response. We know that coronaviruses, they have a much smaller genome, and because of that, a more reduced protein coding capacity. So probably, some of those potential or eventual immunomodulatory properties, they would be played by proteins such as a spike or other major proteins that are produced by these coronaviruses. So it’s playing an important role in the biology of these coronaviruses, such as in infection, replication, or in the case of SARS-CoV-2, promoting acute inﬂammatory responses, such as the still not-well understood and characterized cytokine storm. We are trying to ﬁnd if there are additional roles that these SARS-CoV-2 proteins could be playing in those terms. I think that so far, we found interesting ﬁndings, and we hope and plan to be able to publish it soon.

Barr: What are some of them? Can you speak a little bit even vaguely about what some of those ﬁndings are?

López-Muñoz: Well, I think I will invite Jon if he wants to comment on that?

Yewdell: Yeah, no, I think you can. I think we can talk about it. I mean so it's funny because it goes back all the way to a ﬁnding I made when I was a PhD student in the late 1970s. As I said, I came from a lab where we made the ﬁrst monoclonal antibodies, and we were mostly interested in monoclonals to the equivalent protein of spike which is called the hemagglutinin on the HA ﬂu virus which attaches the virus to cells, so the antibodies are really important in protection. But while we were making them, we also found that we made antibodies to an internal protein called nucleoprotein. That was part of my thesis: to show these antibodies were speciﬁc for nucleoprotein. I was, without going into the history, basically using them as a control in experiments for some other protein, that other viral protein we thought was on the cell surface, and what I noticed was that when I put the nucleoprotein monoclonals in with infected cells, the cells clumped. They stuck together. So the antibodies were somehow sticking the cells together, which in the end was because nucleoprotein is expressed on the cell surface. So one of my papers for my PhD was showing that, hey, there's ﬂu nucleoprotein on the cell surface. There was a previous paper making that same conclusion, but it was done with polyclonal antibodies, and particularly back then, it wasn't easy to make antibodies against a single viral protein. It was always possible that they were against something else that you didn't really know was also in the mixture because you puriﬁed the viral proteins from a virus and so every viral protein was there at some level.

The monoclonal antibody paper was kind of a nice contribution to the ﬁeld, but no one really had much to make of it. At that time, we didn't know what the CDAT cells were recognizing. One of the possibilities then was that they were recognizing this intact protein on the cell surface, and that turned out to be completely untrue—zero truth to that. What the CTLs recognize are small peptides that bind to MHC class 1 molecules, and I actually spent the next 35 years studying that. We're still studying it, and that's what Alberto was studying. But still there's nucleoprotein on the cell surface, and until Alberto came along ,we really had no idea how it got there or why it stayed there. So there it was, and then we had done some preliminary experiments (Walter and I) to look at whether the antibodies speciﬁc for the protein could protect mice, and we didn't really see anything, but years/decades later a couple of groups found indeed that antibodies against nucleoproteins could protect animals against death from inﬂuenza virus.

And then it turns out, as I was saying before, there's all these different kinds of antibodies that we've basically ignored, that don't classically prevent infection, but the immune system in its wisdom and through its evolution has lots of ways of using these antibodies, and one of the ways it uses them is it has cells called natural killer cells that have receptors for the conserved part of the antibody called the FC, and these attach the antibodies to the natural killer cell which can then kill the infected cell or secrete cytokines that prevent an infection. That's probably how the nucleoprotein antibodies are working, and the beauty of antibodies to nucleoprotein, tying back in with the Mali study, is that antibodies against, say, the seasonal coronavirus nucleoprotein are reasonably cross-reactive, much more so than spike with all the coronaviruses. If you're infected with the seasonal corona as we all are every couple years, with a couple of them at least, we all have antibodies against nucleocapsids against the seasonal coronas, many of which will cross react against SARS-CoV-2, and MERS, and SARS-CoV-1. These antibodies are there and they're important. Alberto started studying that, and Alberto it's ﬁne to say what basically you found.

López-Muñoz: Yeah. As Jon was commenting, so far, we've seen that we can ﬁnd this SARS-CoV-2 nucleocapsid protein, which is commonly described in the literature in these thousands of papers that Jon was commenting on before. So far, it's been said that it's not localized in the cell surface, but we've been able to see extracellularly that nucleocapsid protein in the actual context of the real infection. We also are close to being able to explain or to understand the mechanism behind this interaction; how that nucleocapsid protein would be kept and maintained in the cell surface of those infected cells, but eventually it would be able to be transferred between cells that are not infected yet. We performed some experiments, again using flow cytometry in the lab, in which we have transfected some cells with a plasmid encoding this SARS-CoV-2 nucleocapsid protein. We have incubated those cells separately allowing them to express SARS-CoV-2 nucleocapsid, and then staining a different untransfected population of the same cell type, and then mixing or incubating them together for 24 hours. We are able to see similar levels of nucleocapsid protein in the cell surface of both transfected and untransfected cells which would suggest that this ability of nucleocapsid protein to be transferred eventually in the context of infection. Those data are also supported by the actual infection experiments, and we also see when we infect cells with SARS-CoV-2, we then measure them after staining them with antibodies, we also see cells that are not expressing spike (which would be like the canonical marker of SARS-CoV-2 infection), but we are seeing those cells which are actually exhibiting or presenting in their surface nucleocapsid protein. What does it mean in terms of biological relevance? Why is it signiﬁcant in the context of the actual infection? I think that is still to be determined, but I think that is pretty encouraging and interesting. This story we are trying to build up.

Barr: That is really interesting.

Yewdell: That's really cool, and Alberto made more progress in one year in understanding this than I made in 40 years with the ﬂu nucleoprotein. He's done an amazing job and just the whole project was his conception, and all the experiments and everything needed, he's done an amazing job on this actually. I'm really proud of him, so great stuff.

López-Muñoz: I appreciate it. I have to say that Ivan Kosic, he's been playing a critical crucial role in the development of this project like providing key antibodies, trying to help me to understand how to design and how to progress and move forward, all the critical experiments that we've been designing together. So, I would say that it's been a teamwork effort to move fast and try to be competitive and try to shed some light in there.

Kosik: Alberto is the driving force, we said you know. Absolutely.

Yewdell: Everyone in the lab, people in my lab have done an amazing job, because I haven't, you know, until recently—until post-vaccination—I wasn't coming in, of course, being in a highly susceptible age group for serious disease, and I provided some input, but not much. I mean the guys are really running the lab, and then helping each other, supporting each other, and it's been an amazing team effort. Jaro [Jaroslav] Holly has made a great effort and a couple—

Barr: Can you speak a little bit about his effort and some of the others in your lab?

Yewdell: Yeah. Jaro made a whole bunch of cell lines that express the receptor ACE2 and the TPMRSS2 for protease and furan, and he's made lines that the whole institute is using basically.

Barr: That's great.

Kosik: I want to add that he also made the cell line which expresses the spike protein which substantially decreases the amount of the money you need to actually rescue those pseudotyped VSV particles with spike, because now you don't need to use expensive chemical reagents to get that gene inside the cells and then infect the virus. But you can conveniently just infect the cells, and that boosted the titers and the whole workﬂow dozen of times.

Yewdell: Yeah, and I mean what's great is that in our lab, and I think the whole world basically, you know particularly postdocs, they need to have ﬁndings to make papers to make careers so, of course, it’s self-interest driving the work, which I think is good and that's human nature, but on top of that I think the motivation of people in my lab and all around the world has just to make a contribution to this pandemic. People are like, man, this is awful and, boy, I have the skills to do something to help. I think that was really the motivating force for most of the work that's been done. People just wanted to contribute. This has been terrible for humanity, but at the same time it's a huge win for science and for biomedical research and for virology and viral immunology. I mean thanks to all the money invested by the NIH, which amounts to many, many billions of dollars in basic research, we had the tools we needed to deal with the pandemic as quickly as we did. So largely driven by NIH research, we knew within weeks what the cause of the disease was, and there was a vaccine basically ready to go immediately because of decades of research. And all of us who are in this ﬁeld feel very proud of this small part that we have collectively provided to enable protection of humanity basically. I think we're all proud to be virologists and viral immunologists, and whether or not you actually worked on coronaviruses or something similar, it doesn't matter. I mean all this knowledge came together.

Barr: Yeah, deﬁnitely.

Yewdell: It took so many different kinds of expertise to react so quickly to the virus, and people around the world working together, which is another part of it.

Kosik: Yeah. We're sending this matter, these cell lines back to Europe next week, and you don't probably keep track, but we shared these resources—a whole assay set-up including the pseudotyped viruses, plasmids, cell lines—to dozen of places around the United States at this point.

Yewdell: Yeah. I mean one of the things that we've been very proud of in the Laboratory of Viral Diseases (LVD), our lab chief for many years is Bernie Moss who developed vaccinia virus as an important vaccine vector, and one of the things that Bernie's always been about, and all the members of LVD, is when people ask us for reagents that we've published, [then] no strings attached, we just send [them].

Barr: Is that the primary way that you all have been collaborating with others at this time? By sending your assays and so on?

Yewdell: That's one of the ways. A lot of the projects were more involved intellectually. And it works both ways, and people have sent us things as well.

Barr: So how have you all been involved intellectually?

Yewdell: Well, it just depends on the project. And there's a whole bunch of things we started that we're probably not going to ﬁnish where it was a lot of talking together to get the thing off the ground, but that's just—Covid is normal science, just on steroids and accelerated. There's no lab I've ever worked in or known about where every project you start, you ﬁnish successfully. One of the things you have to learn as a young scientist, if you're going to have a reasonably happy life and reasonable career, is that most things end in failure, and most experiments don't work. At least not the way you thought. And sometimes you can make progress, and many times you can't, and you have to start and do something else. That's just the way science is. If you're going to do anything that's worth doing, it's not easy, and failure is your most likely outcome. One of the key things in making it through your PhD and making it through your postdoc is learning how to deal with failure. So okay, it didn’t work. What to do? What do we change? Okay, now what do we change? Now what do we change? Or it works, but you get something that you don't even see how it's going to be a paper, right? So sometimes you can power through it, and sometimes you'll make one of the great discoveries of your career by doing that, and other times you just have to give up, and one of the things that's really hard to learn is when do you give up? Right? And I'm still learning.

Barr: Dr. López-Muñoz with your study do you have subsequent research in mind based on the work that you've been doing?

Kosik: Oh yeah, absolutely. Many. So with Jon you know, this is one of the best lessons I got from his lab, which is don't do obvious things, think smart, and even be patient. And even if it takes time, it will pay off. So that was our initial strategy for the whole coronavirus. We knew at the beginning that there's going to be a lot of papers about the monoclonal antibodies, therapeutic antibodies, vaccination, all sorts. We didn't even aim for that. So our goal is more long term, and even if the impact of those papers one year from now or two years from now, it's not going to be as huge at the time we publish it. It's still going to be the equally important contribution to science, so that's kind of my way of looking at that. Like I want to do the good science, and of course we need to publish or perish, of course all that’s in mind, but it's like we have a project on-going. I'm going to mention one which is also related to ﬂu. I came up with an idea to genetically modify antibodies to make them go inside the cells. So half of all the proteins your cell is making only stay inside the cell, more or less. So when those are all the targets either for cancer or infectious diseases or any other disorders you cannot actually aim [at] with antibodies. So I did this for inﬂuenza; I made the genetic modiﬁcation to one of the internal targets of the ﬂu, internal proteins only expressed inside the cell, never actually really leaving the cell more or less, and now this antibody actually has the therapeutic potential.

Barr: That's really nice. That's really great.

Kosik: And we're doing the same thing, you know. This project is quite developed also for SARS as well. Targeting something inside.

Barr: That's really interesting. So Doctor, you know you've done a lot of educating people about the immunological aspects of Covid. Can you speak more about that? I've heard some of your podcasts and things of that nature.

Yewdell: I had the privilege of going on TWIV twice. I think to talk about Covid, and TWIV, for people who don't know, is “This Week in Virology”. It's a podcast that was the brainchild of Vincent Rancaniello. Vincent does it with a couple of close colleagues, and it's a fantastic resource for the world. Its popularity has blown up with Covid. That's a great platform to talk to lay people and particularly to talk to other scientists. So that's fun. I did two before that as well about scientiﬁc careers. One with one of my children; one of my sons is a postdoc at Sloan Kettering who we actually collaborate with, he works on ﬂu, and I think we did the only parent/child TWIV ever done, where we talk about what it's like as a scientist from my perspective raising a young scientist, which I didn't intend actually. So kind of semi-random, but his perspective from seeing what science was like from the other side. So I've done that. Celine Gounder has got a really good podcast; I did that one. I have a friend who's got a show every week on the Tacoma radio which is a low power radio station that's also on the internet, and almost every other week I've been on with him talking about Covid, not oﬃcially. I don't give my aﬃliation for that one, just as a knowledgeable viral immunologist and talking about the disease, so that's reached, I think, a fair number of people. I've done a couple of other things. At my 45th college reunion, which we didn't have because of Covid, we had a Zoom, which was actually better in a lot of ways because about maybe 200 out of a thousand classmates participated, and we could actually see each other rather than randomly bump into each other on the campus. So we did one on Covid which was great, and then that led to another Zoom meeting I did with a group of women. It's like they have like a club where they talk about interesting things in New Jersey, and yeah, it was really fun.

Barr: Oh that’s so cool. Very different audiences.

Yewdell: Yeah, that was a good one. One of my daughters-in-law works for a pretty large company who was super interested in Covid, and I did a Zoom with them where they could ask questions. I've really tried to share my knowledge with people and with the general public.

Barr: How do you all—this is a question to all of you—stay tuned with the literature and what's going on? I’ve heard a lot of people are going on Twitter now.

Yewdell: 130,000 papers.

Kosik: So one cool way we actually have for this that I really enjoy is we have a WhatsApp group in the lab now, and that's what we use to organize the things in the lab and looking for reagents, because now we're working on a morning schedule/afternoon schedule, so you don't have access to every single lab member all the time. We just text, “Do you have this chemical or that”, but on top of it, we also share cool papers, so whoever is reading some cool paper, he will post it to the group. Then it's kind of like somebody is actually picking the cherries for you, because the lab has a very high criticism so if somebody submits a paper there you know it's worth reading.

Alberto: Yeah, I would say that since the pandemic, like he said, that we got engaged that way, the whole lab. It was a way to keep everything and everybody together and in touch with each other, and we've been having that as a game—trying to ﬁnd a cool paper every week, trying to share it with everybody on a weekly basis, trying to keep everybody engaged in the lab.

Barr: That’s fun. Well, I think that moves me on from your roles as scientists to your roles as people who are living through this pandemic. So what has it been like coordinating schedules and ensuring everyone's professional, educational, and personal needs were being met during Covid when things were very upended?

Yewdell: Well, the lab was kind of self-organizing, because, as they'll tell you, I'm not the most organized person. People did an amazing job in really organizing the lab. Ivan played a big role in that. Another very senior postdoc, Devin Dersh, was great. Jaro played a big role. Everyone really has chipped in. We were able to get back in the lab quicker than most because of the Covid work. And then also our lab chief Ted Pearson was really generous and set aside other space for us. So we've had basically everybody back for a long time now, and as Ivan mentioned, we still have these space limitations. So usually there's like half a lab here working at any one time just to maintain the proper spatial distance. I think people have struggled to various extents, and particularly for the foreign post-docs who don't necessarily have support groups in the U.S. Alberto can attest to that. Thank god for Zoom to at least maintain some contact with people but it's been hard, and some of the postdocs are from countries where it's been pretty bad, and people got infected, and friends or friends of friends have died. And emotionally that's a real struggle.

Alberto: So probably fearing that our family could be infected back at home and not being able to be there to support them, or even sharing some time with them other than virtually speaking, I think that it's been a bit challenging. But, truth be told, as we were saying, we've been having lab meetings every week, trying to be together as a group. Also in the Laboratory of Viral Diseases, here with everybody, once we have resumed the weekly seminars we are doing, that's been a very nice way to feel that you are part of something that is bigger, part of a community, which I believe that as Jon was saying, when we are from other places around the world, it's a critical thing that keeps your mental health and your attitude positive. We want to come to the lab every day and try to keep up every day this sentiment, this feeling of being part of a community and feeling like that.

Barr: Yeah. You've spoken about some of the challenges, like being away from your families and the fear of them getting sick, but has Covid presented any personal opportunities for any of you?

Kosik: Yeah, absolutely. Absolutely. You know, in many aspects. I'm going to say this brought me more deeply back to my roots, to my history of my nation and country. Because of Covid and all the mental pressure, I was looking to start doing something new. I started to play a very unique, very Slovak–it's only in Slovakia–flute called a fujara. It's almost seven feet long; it's handmade. I actually ordered it back in Slovakia and got it shipped back here to the US. And I am not a musician, not an artist, but it’s really making me happy, because this is something every Slovak secretly or openly wants, and the condominium board is okay. I'm going to do something else.

Yewdell: Probably more secret. And what about the whips?

Kosik: And the whips, which is another Slovak thing. I started to do the whip cracking. My nation, hundreds of years ago, were mostly the shepherds, and it was a communication tool. People living on the far away farms, they used the whips in the evening as a signals, and they were whip cracking, and the sound of that ultrasonic boom was actually spreading around the country. And basically there’s a communications and also it has a religion in it a little bit like scaring off the ghosts, so I started doing that as well. Now I have a collection. A year ago I started with one whip; now I have seven including a cool Indiana Jones-bullwhip style.

Yewdell: The NIH police have also been scared away by the whips.

Kosik: I did the demonstration in front of the building 33. And that was you know – yeah.

Barr: That was very cool. I'll have to come and see it sometime.

Yewdell: Yes, you will.

López-Muñoz: Absolutely, you should.

Yewdell: We are a full entertainment center. Exactly right.

Barr: Okay, yes.

Yewdell: I could play my saxophone. Ivan can play the ﬂute.

Barr: All right. Yes deﬁnitely. What is one way each of you have tried to stay positive during this time. I think Ivan answered some of that question but Alberto or Dr. Yewdell?

Yewdell: I'm just a naturally positive person.

Barr: That’s a fair enough answer.

Yewdell: So business as usual, and I have a whole lifetime of exercise regime, and I kept that up as best as possible. I think daily exercise is a really good way to keep your mental sanity, so I was running, riding, lifting weights every other day. I think that helped a lot. Zoom has been enormous help, you know, just being able to see each other every day and family members. A couple years ago, it wasn't very good, the video conferencing, and now, thank God, the internet's better, and I think that's bailed out the world in a lot of ways.

Barr: Definitely.

López-Muñoz: Yeah. I would like to add as a personal note that other than obviously being able to enjoy Zoom and stuff, truth be told, that one thing that has been keeping me in a positive attitude has been the fact or acknowledging that we’re living in a historical moment. And the fact that by chance-ish, we found ourselves, like in my case, in the National Institute of Allergy and Infectious Diseases (NIAID), particularly in John Yewdell’s lab as a postdoc, having at my disposal every resource that I could need to develop my research. Being able to see how, in the building in front of us, a worldwide vaccine, which is saving millions of lives, was developed. Being able to get access to that vaccine, participating in a clinical trial when it was tested.

Barr: Oh, you were part of the clinical trial?

López-Muñoz: I participated. Yes.

Barr: Oh, that’s kind of a really unique, cool experience.

López-Muñoz: It was, but particularly for me it was, because I knew that I was not only contributing to that scientiﬁc milestone, but it was developed in front of the building that I'm working in. I think that all of those things made me be aware of the particular, signiﬁcant, exclusive historical moment that we've been living in and trying to see it as an investment for my future, for my career, for my opportunity, and also feeling happy for, kind of, being able to be part of it somehow. Like, personally speaking, at the bench and doing my own research, but also being part of that whole thing being developed, like participating for example in that clinical trial.

Barr: Did you get the vaccine or the placebo in the trial?

López-Muñoz: Well, obviously I didn't know at that time. But obviously, based on the reading that all of us did about the symptoms, the side effects that people getting the vaccine could have, and I got, and after that I was unblinded, so I was conﬁdent that I got the vaccine in the ﬁrst round.

Barr: Oh, that's really neat. That’s really nice.

Yewdell: I’ll just mention, as well Gabrielle, just seeing Tony [Anthony Fauci] on TV every day was inspirational for us. The post-docs don't really know much of Tony personally, just as this inspirational leader, but all the faculty members at NIAID, I mean Tony's a colleague, and just seeing him up there doing such an amazing job for the public, I mean just being such a fantastic public servant, and he's an incredible person. A couple of times I needed to email him on behalf of some colleagues outside of NIH who wanted to get in touch with him, and often within a half hour, he'd get back to me. It's amazing. I'd see him on TV waiting to be in the press conference, and there he's emailing me. It's like wow, you know. His dedication and just his knowledge, his ability to interface with the public, and his honesty. I mean it's really been an inspiration. I think to most of the American people, and then particularly the people who work at NIAID, I think we're all just very proud to work under his leadership.

Barr: I had a question about that though. How do you all feel at NIAID when NIH or especially Dr. Fauci is being so disparaged publicly by certain sectors of the population and run through the mill? It must be very hard for you all because I know it's hard for me, and I'm not even in his institute.

Yewdell: Well, I think it makes all of us angry, but it's part of a larger trend that we have to ﬁght against, where people don't believe in science, they don't believe in facts. As federal employees, we're not allowed to have political opinions, so I won't express any, but that also as human beings that still complicates it, right? But you know mainly, this is a battle. Science is not something that's natural to humanity. We went, as we're 300,000 years old as a species, we basically went all of that time without believing in science. It's only the last 400 or 500 years, and much sooner than that for most people, where science is really a governing philosophy. This is a battle now we can never give up. I mean there are forces in society who are anti-scientiﬁc, and that's not left wing or right wing, it's both parts. There are crazy people everywhere who don't believe in rational thought, and part of our job as scientists, this is part of what we need to be educating the public on: the importance of rational analysis of the universe. We're scientists; this is part of our job, and if we don't do it who will? Scientiﬁc method is probably the most important discovery humanity has made, and I think it's our job as scientists to promulgate it and support it. Tony is the avatar of all of us—seeing him there, defending science better than anybody, and just giving the facts as he knows it and if he doesn't know something he says it, which is also part of being a scientist. You don’t claim everything, you just say this is the knowledge we have, this is the best decision we can make. So all this is important, and Covid has really brought this out as well as the importance of science education for the general public. Now we're facing people who won't be vaccinated, and this is just mind-bogglingly stupid. Educating people in America and around the world on relative risk is another thing where people are basically ignorant. Even those with very advanced degrees don't necessarily understand numbers terribly well and odds, and really all of us, it's our job outside of when we're in the lab [to] friends, neighbors, colleagues, people we know, explaining the importance of science and the importance of—

Kosik: Communicating the science.

Yewdell: Yeah. And risk management. All of that.

Kosik: I have a good example of this actually. It's a bit personal, but I got appendicitis very recently.

Barr: I'm sorry to hear that.

Kosik: That's okay, thank you so much, but there is a study about the mRNA vaccine, which suggests there is a slightly elevated risk for appendicitis, and that being said, I wouldn't hesitate a millisecond to get the shot again, even if I knew it before. I was not aware of that, and that's about the relative risk. Everybody has the risk of getting appendicitis, but now the risk of getting SARS and actually having a serious medical problem—that's much higher. Also my brother, again bit personal, my brother got a more severe case of Covid. He's 47, and for a couple of days, I was sitting on my cell phone, waiting for a message from him saying, okay, I can breathe now or now I cannot really breathe; I feel like I have something really heavy put on my chest, and I feel like I'm breathing but I'm not breathing. It's important to understand the risk and be able to balance what is really good for us, and there is absolutely no doubt the vaccine is what is good for us.

Barr: Is he much better, your brother? I hope?

Kosik: Oh, he's substantially better; however, it's been more than two months, and he says that he's maybe at 70 percent of his shape, he said he feels like he got hit by truck. Really any physical activity he's engaged in while he's calm, everything is okay. He wants to actually do some physical activity; he says he's still not able to do so. So everybody needs to get vaccinated, because—

Yewdell: Long term Covid, it's very prevalent, and it's really bad.

Kosik: But that's your choice: to actually take things into your hands and control and make sure that this is not going to happen to you, to your family members, to anybody you love, or anybody.

Yewdell: It's an absolute. There's no reason not to get the vaccine.

López-Muñoz: Right. And I would add particularly now here in the United States when you can go to any like CVS or any other location, and you can get an appointment for the vaccine right away.

Yewdell: Free.

López-Muñoz: For free! Which is a privilege that we have here while in other countries they cannot. They have to be waiting for their age group. People are struggling to be able to get the vaccine while here, we have already produced enough to allow everybody [to be vaccinated].

Kosik: Or you have to travel hundreds of miles. Like in my country, my wife's grandmother (and my grandmother-in-law), they had to be 78 years old. They had to travel 150 miles to be at some hospital somewhere in the country at 7:10 in the morning.

López-Muñoz: Absolutely. I really think that this is a way to show appreciation, not only, as Jon was saying, for self-interest, in getting the vaccine, because obviously your health [comes] ﬁrst, but also showing appreciation that in one year, we've been able to have these signs all around the place saying, “Covid-19 vaccines available” for every single citizen here. It's astonishing.

Yewdell: Great stories. One of the great success stories in America actually.

López-Muñoz: What better way to show appreciation than getting your shot?

Barr: Deﬁnitely.

Kosik: Well said, Alberto.

Barr: This is my last question, and it's a thought-provoking question. How do you hope Covid-19 will change the world for the better?

Kosik: It already did, I think. All that science, what we've been just talking about, the progress we made and how the scientiﬁc community tied them up together, sharing even more than ever before: information, resources, everything. Hopefully this will keep going even long after Covid.

Barr: They say that the past 30 years we’re looking at everything through the lens of AIDS or HIV. Do you think that the next 20 to 30 years we'll be looking at things like Covid or coronaviruses or it's hard to say?

Yewdell: Well, it’s certainly going to change vaccination forever. Showing what could be accomplished with the right amount of resources. It's just, again quantitatively, just thinking about things in the right way. How much does the disease cost and how much are you spending to ﬁx the problem? For years, people have estimated ﬂu cost 10 to 50 billion dollars a year in economic costs in the US alone.

Barr: That's a lot.

Yewdell: Yeah, not to mention lives lost. Okay, and what have we been spending on the research to ﬁx that problem: a tiny fraction of that. And what's Covid going to cost the global economy? I don't know, it's looking like 10 trillion dollars. Even if you add up everything, we've spent over all of the history of NIH, it's less than 100 billion dollars over 50 years, and we need to be spending much more money on science and technology. And not just for vaccines, but global climate change is a much bigger problem than Covid, and we need to spend whatever we need to spend to reduce the effects of this as much as possible. If we don't learn the lesson now, I don't know when we're ever going to learn that.

Kosik: Yeah, exactly.

Yewdell: You know some other really positive things that have come out of it, which isn't really science, is that we have a real, we had a perfect, example where we have a test where how effectively people can work from home. We know it's possible now, and the beneﬁts of that are enormous. Both in terms of taking care of families, particularly young parents who have kids at home, but also in decreasing fossil fuel use, and in reducing trips. I think something most people have learned is, man, I really need to go to work to stay sane because I miss people. You realize what's important in your life, and how important is the daily contact with your colleagues, but at the same time, we can have a rebalancing now where people who don't absolutely need to be at work every day of the week, can spend more time at home, and people, what I've been reading of the data, are more productive, not less productive and have this balance now. I think this can get us to a much better work-life balance, which has been a huge problem for young scientists, particularly people who are married or have kids. The demands of scientists are just to be competitive in the world, to get an academic job. It's too much; we're asking too much of them, and you know this is a really good time to reconsider how we're living our lives and factoring in happiness is actually an important aspect in a career. It isn't just about eﬃciency; how happy are people? This is important, and I think Covid has really focused on a lot of those issues, work life issues, and showed us that we can do things differently and still and still be eﬃcient.

Barr: Yeah. Dr. López-Muñoz do you have anything that you want to add?

López-Muñoz: After the captain having said these amazing remarks, I think that there is a very little left from my side to say, other than adding myself to all of those amazing comments that both Ivan and Jon have made. So absolutely. The only point would be I really like the fact that Covid has brought to its rightful place—I would say from my biased point of view—biology, viral immunology, and ﬁnally science. This pandemic has evidenced how science is the key for maintaining our well-being and being able to thrive.

Barr: Well, I want to thank all of you for your time and your service, and I hope that you all continue to stay safe, and your families stay safe.

Yewdell: Okay thanks, Gabrielle.

Kosik: Thank you so much for having us it was a blast.

Yewdell: It was really fun talking to you.

López-Muñoz: Thank you so much Gabrielle.