Molecular Mechanisms of mRNA Vaccines

Hello everybody and welcome to the Molecular Mechanisms of mRNA Vaccines lecture. My name is Kevin Folta, I'll be hosting this today and appreciate that you're here. We know that 2 vaccines have been developed, and one of them to be released later today potentially for use, another one last week. These are based on mRNA, which appears to be a new strategy but really isn't. And we know that vaccinations are what cure people, not vaccines, right? It doesn't matter if you have a vaccine if it's not getting into somebody's arms. So what I would hope for today's session to do is to provide you with more information that you can use to compel others to rethink the disinformation they've heard, primarily through social media. So I'm speaking today with Allison Kennedy will be working with me here today. Allison Kennedy is a student in my— was a student in my molecular biology class this semester who has medical school aspirations, and she's going to summarize the recent New England Journal of Medicine peer-reviewed report on the Pfizer vaccine. So just to restate why we're here, there's a tremendous amount of disinformation out there. And I would like to be able to use this as a way to provide you with the information that can help you push back. And it's critically important, whether you're a science enthusiast, a scientist, or, you know, just somebody who wants to know more about the vaccine, that the information that we'll discuss today is critical to share in venues like social media, that what you learn is going to change somebody's mind and help to come up with greater vaccine compliance. At the same time, it's not that simple. Facts don't change people's minds, and plenty of facts that are out there and people's objections typically are emotional or coming from a place that's not rational, and we'll discuss that as well. And I'll talk about communication strategies that I've used over the years in the areas of places like genetic engineering, climate, or vaccines to help condition people to be able to be receptive to good scientific information. So I do have to give a little bit of a disclaimer and transparency that this presentation is just me acting as a private citizen, not as a professor at the University of Florida. Does not represent the university, does not represent its faculty, staff, or students. I've been discouraged from having public outreach, so I do this on my own time, not as a part of my university appointment. If you have complaints, don't go to the university, send them to me directly. I own a small position in Pfizer, but it's been there forever, and it's— and just putting that out there since, since that's what you do. And I'm not a physician. This is not medical advice. Ellie is not a physician yet. This is not medical advice from her. And we'll be joined eventually at the end by Dr. Asha Burnings, who is a colleague of mine who worked with me in the laboratory for years, and Asha is particularly Adept at discussing issues of immunology, and she'll be available for the question and answer period, and she is also not a physician but is a wellspring of information.

So why are we here?

We know the pandemic is soaring, the numbers are large, there— it's moving throughout the United States and all over the world, but particularly in the U.S., and we know that The ideas of changing behavior, you know, keeping distance, staying home, covering your face, these have not resonated with many of the American people, and there are people who still believe this is an absolute hoax. For these reasons, we were finding ourselves in a predicament in a public health situation that is going to require a more nuanced solution, and that's going to be vaccines. The problem is that just because you have a vaccine doesn't mean the pandemic is over. You have to get it into a substantial part of the population. So we need to have something like 70% vaccine compliance. We're in a situation where we know that 50% or so of people say they will not receive the vaccine or unlikely to receive the vaccine. So how do we change that? And that's where you come in. Your job is to take the information and use this to dispel the copious disinformation that's present online so that we can reverse that and get more compliance. And it's all about building trust. Trust doesn't come from facts. Trust comes from you being an effective conduit and building trust and then having the facts that are that they're seeking, and we'll talk about that at the end of the presentation today.

So why are we here?

Well, as I mentioned before, it doesn't look like compliance is going to be a sure thing, and certainly the excitement around the vaccine has been a little bit infectious, right?

It's been— people have talked about how—

what a wonderful breakthrough this is. Nurses are sharing the photos of them receiving the vaccine. Our vice president got a vaccine. When you see this, this does change the public opinion, so this is really important and we'll talk about that later, but according to numbers from the Kaiser Family Foundation, you can see here on this particular slide, is something like 41% right now say they will receive the vaccine, which is up from 34% just 3 months ago. However, we have 20% of the population that says absolutely not, will not do it. That's down to 15% now.

The people we need to target are the people who say probably.

I probably will get it, probably will not get it. One of those, you know, those folks who are not yet sold on the idea. And so that's where we need to focus our efforts. People who say they will not get it, definitely will not, you will never educate them into getting it. Their decisions are emotional. They come from places of mistrust, of distrust of the government, distrust of regulators, distrust of Big corporations. So you have to understand that and, and don't spend your time trying to change people who don't make up their mind based on evidence. Okay, you can't change the mind of someone who's made decisions not based on it. Let me say that again more eloquently. If somebody makes decisions that are not based in evidence, you will not change their mind by giving them more evidence. Evidence. All right, so, so the idea is here is spend your time with the people that you can change, and those are the folks in the middle of that graph. Other groups are likely to be—

that are likely to be hesitant have been segmented out of that chart. So the chart showed the people who definitely will not get it.

There's a proportion in people who probably will not get it. Who are they? And this chart that's shown here separates them out, and people who are particularly less likely to receive the vaccine are, according to this graph, according to this study at the Kaiser Family Foundation, are Republicans, people aged 30 to 49, rural residents, Black adults, essential workers, independents, men are less likely than women. People who are very likely to get it are kind of the— are folks like Hispanic adults, white adults, urban residents, households that have serious health conditions, people who are 65 and over. So spend your time with those folks who are more likely to be able to be changed. That doesn't mean ignore the other ones. I mean, absolutely, the, you know, the Black community throughout the world, the African American community here in the States, That is a community that has tremendous distrust for very good reason, and we need to reach out to them, and they, and they do need to have ambassadors who are coming from, from their community to help provide that trust. But so don't— I'm not saying don't spend your time on those folks, but definitely understand that there are different reasons for the mistrust that are going to make communication of this particular topic a little more nuanced.

Okay, so the other important table is this one. When you look at that Kaiser Family Foundation survey and it asks you why people are definitely not or probably not going to get vaccinated, you can look at the reasons, and this is what's really important. We can break all of these reasons down into things that we have a chance to influence and things that we can't, and that's what's really important for you as a communicator to keep in mind. Who can you possibly change? And that's a really— that's really important for us to notice. And when you look at this, I think that areas worried about possible side effects, we can help address that. Do not trust the government, make sure the vaccine is safe and effective, I think we can change that too. The vaccine is too new, what do we know there? I think that's something else where we have an opportunity to create some change. And also, I'm worried about they may get COVID-19 from the vaccine. And there's— these are real concerns, and I think these are concerns that we can adjust. People who say that COVID is just exaggerated, that politics has too much of a play in it, they don't think I'll ever get it— those folks have more belief-based, emotional feelings that we probably couldn't change. And so what I'd like to do today is show you how we can address these areas, areas with the blue arrow. by using the science that's available to us, so that's where we'll really focus our attention. So fundamentals of vaccines, I won't talk about this too much, but essentially it's a protein antigen that's introduced to the body, which gives the body the sense that something is wrong from a pathogen, something there that is a signature that ignites an immune response from the body, and this is much more complex than I just made it, but the basic idea is the presence of a, of an antigen. It doesn't have to be a protein, but the, some sort of antigen that then excites the body's immune response to produce antibodies and other types of cellular responses to confront that challenge. So what we'll talk about today is how vaccines are currently made. We'll talk about the SARS-CoV-2 structure and its vulnerabilities, and then we'll talk about mRNA vaccines specifically and how they apply to SARS-CoV-2. And the trials where Ali Kennedy would jump in and talk about the current state of the clinical trials and the data from them. So traditionally vaccines have been raised in chicken eggs or in tissue culture, so this means growing a lot of virus, and you grow a lot of virus, then either weaken it or, or kill it completely, and then use that along with adjuvants to inject into the body to activate that response against those antigens. And I say antigens because when you're injecting an entire virus, a weakened or killed virus, there's a number of proteins there that are involved and that are going to excite the immune response to it. You know, you think of the flu vaccine, you've got a number of different antigens there that are, you know, the H1N1 or H1N5. Those H and N numbers are different antigens that the body can address. So this is how vaccines are traditionally made, and the problem is, is that in response to a new threat like SARS-CoV-2, figuring out a way to culture it, having the scale to culture it, constructing new facilities and protocols to culture and purify it, is a huge undertaking, and that really sets the stage for the mRNA vaccine. So the virus itself, we've seen a million pictures of it this year. The thing to really keep an eye on with this virus is the different places that provide an antigenic signature. So in this case, we look at the spike glycoprotein. It's this pink structure on the outside of the virus which shakes hands with the cell, and upon shaking hands with the cell, cell is then endocytosed, or is brought into the cell, and we'll talk about that in a second, but it's this spike protein, this pink decoration on the outside of the virus, which can serve as the antigenic agent for vaccine design, and this is exactly what they've done with the mRNA vaccines. The thing to keep in mind is that that vaccines— or I'm sorry, viruses— they have no metabolism. They can't reproduce on their own. These require host-to-host transmission, and they exploit your cells to make more of themselves. So viruses, you know, and Dr. Lauria Capua, who's a virologist who I work with at the University of Florida, is— she says, you know, viruses are just viruses. They They are pretty, pretty, uh, they're not terribly inert, not terribly exciting unless they're being transmitted from person to person. So in this way, our behavior really frames the transmission of the, of these pathogens. So it, it doesn't have much to a virus. I mean, they need to use our cells to replicate themselves, and they're exploiting us, they're exploiting ourselves and our behaviors. replicate more of themselves. There's been a tremendous amount of research into this area in the last year, and even back in March there were cryo-EM structures that were being determined of the spike protein. So understanding the virus in its pre-fusion conformation, meaning before it attaches to the cell, that's what we're looking at here. And we'll talk a lot more about this in just a minute because it's the structure of the spike protein protein that makes it an attractive target. And really, this first step in infection is this connection between the SARS-CoV-2 spike protein binding to what's called the ACE2 receptor. And ACE2 is a protein on the surface of mammalian cells that has roles in inflammation and other processes. The lungs are loaded with them. And the ACE2 receptor is where the virus essentially docks, and then, like I mentioned before, after that connection is then brought inside the cell for replication and where the virus reproduces. So if you can disrupt that connection between ACE2 and the virus, now all of a sudden you have a potential therapeutic or another way to defeat the virus. And this is where different strategies have been born to block the infection, and there's several of them that you can think of here. They've talked about decoys, where you make something that has more— is more attractive, a molecule that's more attractive than the ACE2 receptor. Just a higher affinity ACE2 receptor that maybe could not be attached to a cell, a chemical mimic that could bind it and essentially absorb it away from the cells. Monoclonal antibodies do the same thing. Proteins that are targeting those structural domains of the ACE2 receptor— I'm sorry, of the spike protein— that don't allow it to interact with the ACE2 receptor. Small molecules or peptides— this is what my lab does— we're generating peptides which bind specifically to the spike protein and disrupt the spike-ACE interaction. But the other way you can do this is vaccine-elicited antibodies or convalescent serum.

And this was—

convalescent serum essentially says the antibodies that are coming from people already infected. It hasn't turned out to be a very good strategy, but vaccine-elicited antibodies apparently works well, and that's the fundamental basis of the mRNA vaccine. These are 2 really nice pictures. So what this shows you here is a diagram of a space-filling model on the left of the, of the spike glycoprotein and, and the ACE2 receptor, how these 2 things fit together. The binding domain you can see here in blue, that these 2 proteins come together. And so how do we disrupt that interaction? On the right, there's a model, space-filling model, that shows the spike protein on the left on the surface of the coronavirus and how it would potentially interact with the human ACE2 receptor if it wasn't for antibodies that were directed against the binding domain. So now the binding domain can't interact because it's physically connected to an antibody. There's other aspects of immunity to this as well, or immunology too, that have to deal with the antibodies and what they mean to the virus in the context of cellular immunity, but this is a physical blockage, and that is what, that is, that is, that is what they're going to key off of for this particular approach. So I see I might have a question out there, but I'm unsure the easy way to get to it here.

Let me see.

Well, I'll come back in just a second here. Okay, so the reason this is so important in a pandemic is because RNA-based vaccines, they give you an opportunity to rapidly produce a new vaccine. When you have a new threat, you can move quickly. It's not like you have— not like what I mentioned earlier where you have to come up with culture strategies. This is as simple as producing an RNA, and we'll talk about what that means in a minute, but generating a piece of genetic material that encodes the antigen you wish to attack with antibodies, in this case the spike protein, and it works really well. The Moderna vaccine— and Moderna is one of the major companies in the space— they were able to produce the first evidence of a vaccine in days. As soon as they had the sequence, which I think was January 11th or 20th of this year, the coronavirus DNA sequence was made available, they had a vaccine prototype in a couple of days. They were injecting it into people after 42. That is amazing speed. Nothing like this has ever been done before and in that rapid of a timeframe. So in order to understand how these work, we need to talk about the central dogma of molecular biology. And for many of you, this is old stuff, but basically the idea that DNA is the master, master storehouse of the blueprint of information of the cell, and that, that information through a process called transcription is specific genes are copied into RNA. The information in DNA is copied into RNA, and then RNA leaves the nucleus, leaves the safety of the cell's nucleus, goes out into the cytosol, the rest of the cell, and where it marries something called a ribosome. Which is essentially a center, location of protein synthesis. It binds onto the ribosome where the protein is synthesized. So I think of it often as the hard drive of a computer. The master information locked in the box that you never see is safe and put away. The temporary format of the USB drive, which carries a little bit of that information somewhere else. And then ultimately going to a printer where you can create some sort of output. And you think about that, it's kind of similar to what happens inside the cell. You have master information that's copied to a temporary format that then goes to another place, another machine where outputs are created. The most beautiful part about this, or an important part of this, is that that intermediate step of the USB drive is horribly temporary. And if you've ever seen the lint trap on my dryer, you get about 1 or 2 of these a week that, that don't survive the clothes washing process. It's a temporary format for information storage, and that is RNA. RNA carries that little bit of information out into the cell to be able to produce a protein, yet it's a temporary storage format. So when we talk about RNA-based vaccines, scientists or pharmaceuticals are generated— scientists generate pharmaceuticals that are RNA-based. You take the RNA and essentially bring this into the cellular environment, just the RNA, and then that RNA goes to the ribosome where you create the protein. So you don't have any hard DNA storage. This is just the intermediate that's introduced to the cell. and then creates the protein. In this case, the RNA that has the message and the DNA— I'm sorry, has the RNA that has the genetic information for the spike protein that then is translated by your cells to create the final protein, which is the spike protein antigen. So you can see a lot of good advantages of being able to do it this way. We'll talk about that in a second, but one, one important part of this that, that we really haven't heard much about in the news is that RNA itself can induce an immune response. RNA complexes with something called toll-like receptors, and this paper from 2005 was very important. This is from Katalin Karikó and Drew Weissman together at the University of Pennsylvania. They determined that you could change the types of bases used in RNA, the chemicals used in RNA, to be able to make it invisible to that surveillance mechanism, and this was very important. This was an extremely important development in the discussion of RNA vaccines, because now you had mRNAs that could be brought into the cell and be trans— and be translated into the protein antigen Yet evaded the body's natural mechanism for responding in a negative way to foreign RNA.

Very important.

And Dr. Karikó has an excellent story if you can read about what she went through to do this. She's been studying mRNA vaccines since the early 1990s and had a real tough ride of it because people didn't think it could work, and it does. Robbin as we know now. So congrats to her. How do you get it into the cell? How do you get it into a ribosome? So they take the RNA and it's encapsulated in something called a lipid nanoparticle, so LNPs, and these are nothing more than lipids. Sometimes they're complex with specific cellular, or as well, specific protein signatures along with them, especially in delivery of cancer therapeutics, a little bit different kind of mRNA vaccine, but in this case the mRNAs are inside a small little bubble of lipids or fat molecule basically that complexes with the cell, is absorbed into the cell, and then delivers that RNA to the ribosome where the protein is created. So when you get the injection of an mRNA vaccine, it's complexed in these lipid nanoparticles, that go into the deltoid muscle of your arm. They find homes in those little inside your cells of your arm, go in there and produce the protein. The protein is then, then decorates the outside of those cells and is inducing an immune response. So that's really how this works. It's a pretty simple process in general. The strengths of this are really important. It's that the body is exposed to a single component of the virus, so a single antigen, not multiple antigens. For the company, it makes it easier to manufacture, less expensive, very flexible, makes it very easy for them to scale up and scale down, fewer processing steps, standardized steps for all mRNA vaccines, so you can be producing a SARS-CoV-2 vaccine today. You can switch your pipeline to a different virus tomorrow. And that's huge because there are many of these coronaviruses in natural repositories that have the possibility of becoming jumping, becoming zoonotic, jumping species, and becoming threats to humans. The other nice part about this that people actually frequently worry about is that it's going to interact with their DNA. And we know that the RNAs that are introduced do not interact with DNA. They don't even enter the nucleus. And the most important part is, like that USB drive of the cell, the response is transient. You generate the transient mRNA that, that then can— that then is degraded by the cell. And we know that the half-life of mRNAs in these cases is on the order of hours. So So a very short half-life, and it is a transient response. The downside to mRNA vaccines is because they're typically very unstable, and you have to keep them frozen or cold. You've heard about the Pfizer vaccine that has to be stored at -100 Fahrenheit. That is one significant drawback of this particular technology. Again, the idea that there is a potential inflammatory component. We know that there are a lot of unknowns about the long-term durability of the response, if the antibodies stick around, if they're— if the response is somehow neutralized and not long-term durable. All of these things are not known yet. Even some questions about safety, long-term safety. Everything appears great. But those are all questions to be answered as time rolls on. Right now, it seems very safe, and we'll talk about that in a moment. So the absolutely horrible name of Operation Warp Speed, which gives the idea that this is maybe not a rigorous process, yet it is. It's a public-private partnership, and the goal was to overlap regulatory steps, still go through the regulatory circuits, but overlap them and do them together rather than make them iterative, you know, one after another, which can take billions of dollars and years. $10 billion was allocated to multiple companies to take on this problem. Different approaches were taken on. The mRNA vaccine was one type, and there are a number of companies in that space. Also an adenovirus-based vaccine, self-amplifying RNA, monoclonal antibodies, regular weakened viral strains, all the different strategies were taken on and funded and are gradually moving through the process of testing and approval. And this is what's really important is that the public says that this was too fast, not enough oversight, one of the major reasons they're concerned about it, and for us to convince them that this is something that's reasonable, we have to remind them that the full regulatory process was engaged and yet was done in a way to limit the bottlenecks and restrictions. And the idea here is to be able to, to test for whether they work or not. Let's fail fast. And so far, things have worked very well. So it's a normal regulatory process, just overlapped and condensed. The other thing that people expressed in that earlier survey is that mRNA vaccines are too new, new technology, untested and unproven. The part of the thing is though, is that scientists have been working on this since 1989, that really the first mRNAs were injected into the abdomens of mice back in the 19— I think 1989 and 1990, where you could— they would inject RNA for firefly luciferase, the protein that creates the glow. and that when you would inject that under the skin, you could use highly sensitive cameras to see the illumination in those tissues. And there has been substantial change, progress in mRNA vaccines. The big breakthrough, as I mentioned before, was Karikó and Weissman in 2005 with making RNA invisible or less visible to the body's surveillance system. It's been said that this is unproven technology that has never been used before. never passed regulatory. That's not true. There's some things going around online that we've had to debunk, so when you see those, it has— there has been an mRNA vaccine on the market, something called Onpattro, since 2018, and it corrects a rare hereditary peripheral nerve disorder, so this has been proven technology. It's being used in other contexts all the time. The pipeline is extremely vast, and I think this is Moderna's. As you can see, a number of mRNA-based products that are moving through Phase 1, Phase 2, and Phase 3 trials, so certainly not just a one-trick pony. SARS-CoV-2 vaccine was something that has become their, their spotlight product because of the pandemic. Other country— other companies in that space, CureVac has this COVID-19 vaccine, rabies vaccine, yellow fever, a whole bunch of other ones, Pfizer, AstraZeneca. There's a— so there's a number of companies that are playing in this space and, and are producing vaccines. The slide on the right shows the number of projects that are underway in In mRNA vaccines, many different products that are currently being tested or available that will be soon dispersed in a number of countries through academic, public, and government laboratories, so this is not a new technology, it's not an out-of-left-field technology, this is something that's very well understood, That has been used and is being used in many different formats. It's just that the Warp Speed opportunity to be able to condense the regulatory process is what sets it apart and makes it unique. Traditional vaccines go from lab and animal studies through Phase 1, Phase 2, and Phase 3, where we really question different— ask different questions about safety, Dose and response, dose response as it applies to side effects that may happen. The public again says that this one came up too fast, but it went through all of these different steps. And really Phase 3 is where we'll focus on today. What did we learn from Phase 3 in that development process? This is where I'll turn this over to Ali Kennedy. Ali was a student in my course, um, in my molecular biology course, and she is, um, is a future medical school student here, and she's, uh, taken on the paper that was recently published a week ago that talks about the safety and efficacy. So go ahead, Allie.

Absolutely. So when we're looking at the Pfizer paper based around the safety and efficacy of the COVID-19 vaccine, It shows some really wonderful results, and it had a pretty wide diversity as far as people that were enrolled. We see a really good spread of race. We see a really good spread of age. And I think really importantly, we see a good spread of different health conditions that are represented in this study. I think there was something like 600, 700 HIV patients. We had patients with hepatitis C. We had patients with hepatitis E. And so looking at comorbid conditions was also taken into account in this study. And when they were looking at this study, they wanted the minimum efficacy of 50%, and what they found really blew that out of the water. And you— it's a 2-dose system, so after that first dose, even if you didn't come back for the second dose, you would still be showing around 50% efficacy.

Mm-hmm.

And after that second dose, they show about 95% efficacy across most groups. And They had a couple of different expectations that were obviously exceeded in this trial. Another important fact about this trial is that it wouldn't have been able to be released as quickly. Phase 3 wouldn't have been completed as quickly if it weren't for the fact that COVID-19 is rampant in the population. So they needed to have a certain number of events, at least 150 confirmed infections, in order to complete the study. And they got that quickly. So this started in about April. and they were able to complete it by now just because of the sheer number of patients that, that were getting COVID-19 throughout this time. So you can go ahead to the next slide. So this is the data showing the efficacy, and you can see that certain groups are too small to really say, okay, in groups over 75 years old, we can say that it's 95% effective. But across the board, even in the largest populations, it's showing Pretty strong efficacy. And I think it's important to bring up for people that are on the fence maybe about mRNA vaccines in general, as opposed to attenuated vaccines like flu vaccine, is that this does absolutely exceed the, the efficacy of any yearly flu vaccine. So the flu vaccine is pretty lucky to see 60% efficacy, and we're seeing pretty consistent 95% efficacy. in this vaccine, and that's after that second dose. But Moderna also found similar, similar results to Pfizer, so it's a really compelling argument towards this type of vaccine, and it's really promising actually because when we're looking for 50%, we get 95%. Obviously, that's a really wonderful finding, and, uh, and it's across the board. So every age group is represented here from 16 to over 75, and it's, it's a, it's a really lovely finding. You can go ahead to that next slide. So here's where it gets a little bit hairy, and I think this is a really important thing to be able to recognize and talk to people about, is the occurrence of side effects. So here we can see that in that first, in these graphs here on the right, let me move my little chat bubble. That in both groups, over 55 and under 55, there was a pretty— there was a fair occurrence of pain at the injection site, although it wasn't super significant. It was something like, I think, was it 4 or 5%, Dr. Folta, of patients were experiencing pain at the injection site?

Yeah, I think it was, it was pretty low. Yeah.

Yeah, yeah, but it is significantly more still than you experience with the typical flu vaccine, so it's something to think about. You do experience— there does seem to be a greater incidence of side effects in this study. And those side effects include pain at the injection site. There's also redness, and there was swelling, but those are the smaller bars. The most commonly reported immediate side effect was pain at that injection site. And if you go to the next slide, we can look at the more systemic side effects that were also reported, which is another really significant thing too. understand and be able to talk to people about. And those top side effects were fatigue and headache. And it was still a really small percent of the population that experienced this. It was 3.8% experienced fatigue and 2% experienced headache, but it's relevant. And again, it is higher than what we see with the flu vaccine. But when we are looking at extremely severe side effects, which we'll talk about in the next slide, there was no difference between the placebo and the administered group. So that's something that's comforting about this. We can see clearly that there's not any severe side effects in this trial, which looked at over 40,000 people were involved in this trial. So it is a really significant finding that we're not seeing any extreme side effects, but people will be really concerned. I think a primary concern about people that are on the fence about getting vaccines, especially ones that are mRNA-based, because it— though it's not a new technology, it's unlike what we get in our seasonal flu vaccine. So, but we can say definitely that it's a lot more effective than we see in our seasonal flu. It's a wonderful percentage. 95% is just incredible. And we can say that although side effects are slightly higher, we're not seeing extreme side effects. in these clinical trials. So it's, um, this was a really great study. And if you go to the next slide, we can see that Moderna, um, and by the way, all of these findings are very, um, these, these were just published, I think, just a week ago. And, um, we know that Moderna had very similar findings, um, around 95% efficacy. So we know that we can prevent COVID vaccine, uh, COVID-19 through these, this 2-vaccine system. So you would have one vaccine, you come back a month later, get that second vaccine, and you're 95% protected from COVID-19. And that, it's a really great finding. All subgroups seem to show similar patterns, but like I said, there's some difficulty in saying, okay, for sure we know it. There were too small, too, too little people getting sick in groups like people over 75 to say that it will— in this study it said, you know, this is 100% effective. Because only 5 people got sick in that subgroup. But across all subgroups, we can say pretty confidently that 95% is the number. So those side effects are going to be important talking to people that are kind of particular about it and are on the fence about it. I think being transparent about the higher incidence of side effects will be, you know, an important factor in all of this. But we're really happy to not see severe side effects.

Okay, thank you, Ali. That was great. Appreciate it. So really what we're up against now is what is the duration of protection that we will see? Will this be durable? Will this be something like the seasonal flu that we need to continually reinforce? Eventually placebo recipients in these studies will be unblinded and will receive the vaccine, so The study group in terms of side effects and long-term consequences will be— have significant more power. We'll be able to test or follow long-term consequences. The big question is, is how do you deploy this now? Who gets it and in what order? And should this be something in the USA, or how do we get this to the entire world, especially the developing world? storage and shipping situation. Other strategies are coming and hopefully will be very useful around the world. And how do we— really, our big question goes back to those primary slides. How do we convert the unsure? How do we get people who are on the fence about this to be excited to roll up their sleeve and receive a vaccination? And that's really the last couple slides here. This is your role, and we talk about the vaccine and how it works and all the stuff that scientists worry about, right? How and what. But when we're trying to convince the public, we need to work with why. And I find that it's very easy for me to help adjust those folks who are in the middle, who are on the fence, when we talk about why it's critical. And really what it's about is about protecting the most vulnerable. Most of us, you know, if we're, you know, if we're not old, if we're, you know, not sick, if we have a relatively good BMI, would likely survive an infection. Some people don't, even, even in best of health, but in general, the vaccination protects the vulnerable, and that's something all of us should aspire to do, that we limit the stress on the healthcare system. When you look at what's happening now, where certain municipalities are running out of hospital beds, the incredible high cost of doing this, healthcare workers who are being exposed, who are burning out or even becoming harmed themselves, you know, vaccination limits the stress on that system and protects them. The other big issue is that it decreases the amount of virus that's circulating in the community. That could translate to mutation, and we already know that there are mutation subpools that are present, maybe a new one coming up in Great Britain. Certainly the East and West have different subpools of the viral sequence, and we don't need something that's going to become more virulent or spread easier. So limiting the amount of virus in the population does help contain that possibility. It decreases the frequency. The other big part of this is that small and local businesses are suffering, and some towns, 25% of their small businesses have closed and probably will not come back. The economy in general will be slow. Some aspects are doing great, you know, your Amazons and your Home Depots are doing well. The rest of the businesses are not. So it really is important from a personal standpoint to a national and community sense of responsibility for us to become vaccinated. The big place where this is being discussed obviously is in social media, and the disinformation is crazy there, so it's important for us to engage in social media in those values-based conversations. Again, you're not going to convince the people who say they will never get the vaccine. Our job is to have polite conversations with those who are on the fence. And remember that the internet is a spectator sport and that the more you discuss this in a reasonable and rational way, you will tend to change hearts and minds. One of the major problems with the internet is that social media provides a way to tie together rare and adverse effects. So in other words, we had a slide up here that showed that during those trials with Pfizer, People died during that trial, but they were equal in the vaccine and placebo group. During that duration of that trial, for every 10 million people, there are 1,700 people who would have died from whatever cause. Now imagine if you got the vaccine on Monday and then that person died— someone got the vaccine on Monday and died on Wednesday. Their family may make that correlation and say, must be the vaccine, right? And, you know, we know that those events could be uncoupled, are likely uncoupled, and had nothing to do with each other. But when someone says, my uncle got the vaccine on Monday and died on Wednesday, and they say this on Facebook or on Twitter or Instagram or wherever, and then someone else has the same experience— people who died just from natural causes, nothing to do with the vaccine— all of a sudden the internet and social media begin to assemble these accounts of rare events and make it seem like it's a trend. And this is why it's so dangerous. It's why it's really important for us to be present in social media discussing what this is and what this isn't. Share the good news. And I showed some slides over here on the right from Twitter of people who are using the hashtag #GotTheShot. Show that this is a community effort. Bring people— get people excited about being part of the movement, why people are getting vaccinated, and it's really important. And a really funny one that I saw today, I thought this was fantastic, from Dr. Ali Khan: Legit goosebumps for the COVID-19 vaccine. I don't ever think I've felt such soaring joy before being punctured. It's no ordinary puncture. It represents the best of human ingenuity, scientific discovery, and deep-seated hope. And, you know, I get a little misty when I say it because I really do love that science has done this, and I really am excited about the fact that we have to— that we are able to share this here today. Um, okay, so I'm going to wrap up there. I really do think we have a moral obligation as scientists and science enthusiasts to step into those positive— into those public conversations and share the stories of how this works and why it's important. And I think that's really, really important. And that's why I'm excited about doing this here today. So thank you to Allie Kennedy for joining me. And we're really happy to take any questions. So if you have questions, I'm gonna try to unmute everybody here. It may get a little busy for a minute. If you, let me see, how do I unmute everybody?

And I did see some questions in the chat as well.

Yeah, okay, so why don't we start with those? Can you read them out loud?

Oh yeah, absolutely.

Thank you. I don't have my eyeballs on.

All right, let me scroll up to the top. So the first one says, if you get the vaccine and are exposed, can you still carry it and pass it along to someone who's unvaccinated? I'm thinking in the direction of unvaccinated children until we know if it is safe for them, Megan asked.

Yeah, unfortunately, I think the data are still out on, like, so for instance, if you received the vaccine and were breastfeeding, for instance, you know, do the antibodies travel through? And I would assume they would. But whether or not they would be in sufficient quantity to be able to impart an effect is hard to say. I think, you know, all of those— all of that work is being done now and being studied now. Children were not included in these studies. They tend to be more impervious to the virus, so those studies probably will commence shortly.

Kevin, I can actually address that because that did come out in the study.

Okay, good.

There is data for that. So a couple of points I wanted to make. Hi, my name is Asha Bruininks and I teach microbiology, so this is kind of my purview. A couple of things I wanted to address about the, you know, about the speed of the trial, about your concern about safety. I want you to know that normally vaccine trials take in about 3,000 participants, okay, and it takes years to gather enough participants to get all that data. In this case, we had 2 trials of more than 30,000 participants. This is the equivalent of 20 normal trials. Okay, so that there are— although RNA vaccines are new, we do have DNA vaccines. They've been used for years in horses, and there's an effective Ebola vaccine. So the technology isn't that new. And then the other thing I wanted to say regarding your question. There are 2 kinds of immunity that you can get upon immunization. There is non-sterilizing immunity, which protects you from getting sick, and sterilizing immunity, which protects— that prevents you from spreading it. Okay. And so what we want is we want that sterilizing immunity to prevent us from spreading it, because otherwise what you get is you get all these asymptomatic carriers, right?

Yes.

So people would get infected, don't get sick but can spread the virus far and wide, which is probably the opposite of what you want. So to see if there's going to be sterilizing immunity, we look for particular kinds of cells in the immune response, and they're called CD4 T helper cells. And they are the ones that are going to activate both arms of immunity to make antibodies and cytotoxic T cells. And the evidence suggests that the cytotoxic T cells may be more important than antibodies to protect against COVID-19. Okay. So there is evidence that CD4 T helper cells are being activated. So that would suggest that this might be sterilizing immunity. And then the other thing is that the immunity that you get against the vaccine is actually more effective than the immunity That you can get against the natural infection, and you ask me, are you crazy? How is that possible? And so it's possible because of the way the RNA is made. The a normal spike protein, you know, Kevin, you showed that picture of the shape of the spike protein, and I don't know if you can. So normally, the the The spike protein has— doesn't stay in that shape all the time. It actually has this really weird thing where it folds out super long and narrow. And normally the spike protein on that virus goes in and out and in and out and in and out. And so it's really hard for the body to make an effective immune response against something that's constantly changing. Now, the RNA vaccines is made so that it is stabilized in this pre-fusion form, in the shape that Kevin has on that picture here, so that your antibodies are going to be against that guy, and that's what you want. And this is going to result in better immunity than you can get against when you're naturally infected against this. So that's, that's why it's possible. And so, um, to answer, um, whose name is that, um, Megan, yes. To answer her question, the evidence suggests that this might actually give us sterilizing immunity and that this would be good. The data is— it is still a little early, but we have some encouraging news.

Yeah, that's a good point about the pre-fusion form of the virus that they were able to use. They mutated a couple bases and changed them to amino acids to change them prolines, and it locked it in that one form. And then that's what they used to create the RNA that would induce the antigenic response against that open form. So yeah, cool. That was a good point. Real good point. And do we have another question?

There's a ton in the chat. Okay. Yeah. Do you want me to keep reading?

Yeah, why don't you go ahead and read them? They're hard for me to see.

Okay. This one is asking about the recording being available, where they can access that. Yeah, we'll make it available somewhere on YouTube Okay, the next question is asking about what was the half-life of the RNA in the vaccine?

That's a good question. Asha, do you know that one?

So no, I don't, but RNA is generally not very stable, so we think it's about hours. Yeah, the thing is, is that the RNA vaccine is not only taken up in the actual cells, in the muscle cells, right? So you inject it in the muscle cells, and because it's in this lipid, lipid sort of envelope, it will actually fuse through the membrane. If it was just RNA, RNA can't get through the membrane, but the lipid can. Um, but more importantly, actually, it is also gobbled up by dendritic cells and macrophages that are underneath the skin, gobble up this protein, and then they will make it and display it on their surface. And those antigen-presenting cells are the ones that are going to be activating the immunity. And so it doesn't have to be there for very long. So these cells, this is their job. Their only job in life is to gobble up pathogens and activate the immune response. So any of that RNA that's going to be, going to be gobbled up will be displayed on those antigen-presenting cells, and that will kick in both arms of that immunity, so both the antibodies and the T helper cells. So even if the half-life is only a few hours, that's enough. The antigen-presenting cells are on the site within 1 hour after the puncture, and they're already there gobbling it up.

Very good.

Yeah.

Yeah, any other— what other questions?

This next one is asking, are there any major differences between the Pfizer-BioNTech and Moderna vaccine?

There are different— I believe there are different parts of the different sequences that obtain a similar protein. I don't know the answer to that precisely. I've tried to find that information.

Yeah, I was hoping—

It's proprietary. They're hiding it. It's, you know, it's their trade secret. So this information is proprietary. They're not sharing it with us.

I was hoping— you asked about doing the write-up about the Pfizer vaccine, doing like a little summary with the data. I was hoping maybe I could do a compare contrast about what they have published on those vaccines. Maybe I can write that up later.

Sure, that sounds good.

Okay, this one says, what's the best way to share this info with my non-sciency friends using, say, the zombie apocalypse? From Jim.

Well, you know, it's— I think the big trick to changing anyone's mind is, and this is something that we learned as scientists, we're learning as scientists, I learned a long time ago, is that facts don't matter and you can't change people's mind with— by burying them in more data and showing them graphs. We did it here today with you because we assumed this audience as science enthusiasts who are wanting to learn more about the topic, but when you're trying to change a friend or family member or someone who may be not as a science adept, you really have to talk about your values. You know, why is this an important strategy? And it's really more about protecting the vulnerable, restarting economies, returning to normal. A fast, easy, inexpensive, adaptable, safe way to get back where we need to be. That's what moves the bar. And so use those strategies. And then if they want to know more about, you know, what bases were mutated to be able to create the, you know, open conformation of the, you know, spike protein, then you can share that with them. But, you know, start with the things that are important to everybody.

So Jeremy says— this is just a comment— but he says, as an employer, I would love to have this presentation for my staff. I don't know if these are publicly available. I guess they're on YouTube, aren't they?

Well, we'll make all this available, Jeremy, but, you know, reach out if we can be of assistance. I'm glad to do the presentation for your group, no problem. You know, this is all about— this is about creating a change through communication and something that I feel all scientists have a moral imperative to do. And I would be happy to talk to your group. So just reach out through my contact information and we'll do that for you or your group anytime, okay?

And to take away your thunder, Kevin, but there's also, if you go to the website of the American Society for Microbiology, they have a program called Ask a Virologist, and that means you can find a virologist in your area or from everywhere, and they'll do a Zoom meeting with your group and, and talk to you about the ins and outs of everything. I did, I did a couple of those for my students this semester, and they're really Excellent.

Very good.

I didn't know that either. That's awesome.

Yeah, it's called Ask a Virologist, and there's hundreds of virologists all over the world that have volunteered to share this, even in different languages if you want it.

That's great. Tiffany asks, what evidence can we give people that supports that the vaccine is safe and likely does not have long-term negative side effects?

Yeah, unfortunately we don't have those data yet, right? But we can only talk about the short-term side effects and the fact that the The inducing molecule, the RNA itself, is a transitory treatment, so there's nothing hanging around after those first proteins and the other cells that have obtained cellular identity based on the antigen. Once those are created, those are the durable agents, and right now we don't know of any specific effects. You could imagine that maybe if there— well, right now it's hard to say, it really is hard to say. But that is an important caveat. But right now, it doesn't look like there are any long-term effects based on, you know, the several months of data that have come in.

Since April, isn't that right? So—

Well, those were the first ones.

Yeah. Yeah, and based on also early side effects like aching and, you know, muscle ache and itching or whatever, those are normal parts of the immune response. Okay, this is the good thing. That means the phagocytes have moved into this area and they're starting this adaptive immune response. So early side effects, I think, I think of them as, you know, indications that we're on a good path to, to activate the immune system. So I think that's the long-term side effects. Although we don't have data for this, we do have data for, you know, the existing nucleic acid virus vaccines, and we haven't seen any long-term negative effects of those.

Yeah, that's true. I mean, some of these have been in trial since 2013. Some of the other RNA virus— I'm sorry, the other RNA-based vaccines. So that's 7 years of data now that they've been in clinical trials for safety. And so, so far so good.

And in horses even longer, although they don't talk to us about their side effects.

All right.

They're in stable condition.

Oh, and I guess an important point I wanted to address against Lauren from Arizona— Lauren Nailing, Arizona, sorry. So these are natural exposures. So it's considered highly unethical to take people and purposely expose them to the virus.

Mm-hmm.

Although some people have offered that they would be willing to volunteer for that, that is not the way it goes. These are natural exposures. So these people are still told, told, go out in the world, do what you normally do, protect yourself. But as we know, even no matter how good you protect yourself, you know, you let your guard down for your cousin or your neighbor, and before you know it, you have COVID-19. And so these are natural exposures.

All right. So continuing in the chat, Chris and Megan are thanking us, and thank you guys for being here. And Andrea is thanking us as well. Thank you, Andrea. Let's see. Lauren asks, in these trials, how is the vaccinated group exposed to the virus? To others who test positive, I'm wondering about the logistics. Thank you for the presentation. And I think, Asha, you answered that.

Yeah.

Just then. Let's see. And then Miss Gaetano says, as a high school teacher, I'd love access to this too. And like they said earlier, we've got Ask a Virologist, and if you want to reach out to Dr. Folta, I'm sure we'd love to do a presentation with you guys too. I think it's really important for high schoolers to understand and be on the cutting edge of this stuff too. Well, I saw that you're from the Chicago area, and that's where I'm from too, so.

What's that?

I saw that Dr. Folta was from the Chicago area, so I thought he might be willing to zoom in on a presentation up in this area again.

Oh, you bet. Yeah, I grew up in Downers Grove and in Chicago and did my graduate work there, did my PhD work there. So—

I teach at a high school about 10 minutes from Downers Grove. I'm out in Bolingbrook.

Oh, sure. Yeah, I know Bolingbrook, of course. Yeah, no, cool. That's great. Well, thank you for being here.

Let's see.

And Sarah says it would be amazing to work with you, Kevin, to offer this type of package for business audiences. So another thing that you could reach out to Dr. Folta about. Okay, here's Marianne asking, how would you recommend approaching very understandable fears that some people of color have, especially since data from the presentation showed only 7 Black trial participants? And actually, I just want to say that that was only 7 cases of COVID that were caught in the, in the Black participants. So it was 9% of the entire study was, um, was people of color. Uh, let's see, I'm sorry, Black people. There was 29% Hispanic, I think, as well. And this says, I know we can be quite confident that there would not be any significant, significant difference in how these vaccines work across groups, but others may not see useful data here given a history we should acknowledge of unethical experimentation. Absolutely. And even using vaccination as a cover for U.S. military operations.

Well, the, uh, yeah, the Tuskegee Experiment and, and others have left some very deep wounds in trust, um, with the African American community here in the States, um, and, uh, in many other instances.

In the case of Henrietta Lacks, there's a ton of distrust in that community, I think, absolutely, and, and, and there's, you know, and I think that the distrust is, is very fair. I think that all we can do is like hope that people are on the fence and that people are open to looking at this in a different light, but I fully understand that hesitation.

Well, I think the other side of this too is that hopefully if anyone in this audience is African American or could, because you will automatically garner more trust with those communities, and that's just known. When hopefully you can engage more with that. That's why it's so exciting to see the nurses on the front lines who frequently, you know, are getting— who are African American are getting these first shots in very visible ways and speaking about it. That does build trust. It does build trust when we are speaking with our trusted communities.

And thanks for bringing it. I think that's a really important point too. Someone asks, I'm not from a science background, but you mentioned at the beginning about the vaccine being grown in a tissue culture, if I remember that correctly. What type of tissue culture is used for that?

Well, this one is not grown in tissue culture, and there are a variety of cell lines that are used to raise human— or raise viruses for therapeutic application. I'm sorry, for prophylactic application. Asha, do you have any idea which ones they use?

So for Let me see. Am I—

Yes.

Yeah, so to replicate, so we do need to replicate the virus in, in the lab to do research, right? And so we must use cells that have that ACE2 receptor. Otherwise, it doesn't work. The virus can only enter cells that have that. And so typically, these are cell lines called, you know, PCL6 that are, that express those receptors. And so they're typically derived from respiratory cells. Not all cells have this receptor, and therefore the virus can't infect all cells. That's— that is what we call tissue tropism, where the virus can only infect cells that have this particular receptor. And so we typically use respiratory cell cultures. But the vaccine is not grown in culture at all. It's made by a machine, right? It's RNA made by a machine wrapped in a little lipid. It's just a machine that strings nucleotides together. There's no tissue at all involved in that.

Thank you so much for that. That's new to me too. That's really interesting. And that's the end of the questions in the chat.

Okay, that's perfect. Yeah, it looks— so just to go back to the very first slide then, just for what it's worth, because since we started the recording late, I wanted to mention couple of things here. These are my personal views and the views of Allie Kennedy and Dr. Asha Bruininks and don't represent the University of Florida faculty, staff, or students, something we always have— I have to say when I do any kind of public presentation. I also do have a small investment position in Pfizer, but I've had it forever and I couldn't tell you anything about it other than I have it. And that I'm not a physician, I'm not giving medical advice. We're just talking about the molecular biology and cellular responses to a virus and how it works and providing communication strategies. So that's what we do. Any other questions or concerns? Any other thoughts before we sign off?

We had 2 more pop up in the chat.

Okay, sure.

This one says, who are the people getting funding for communication actions? I'm sure we're taking a multiple-pronged approach— federal, state, university, etc. CDC has some type of role. I'd love to know who is the most accountable/responsible.

Yeah, it's really hard to say. I don't know who's funding any kind of communications efforts, but they're potentially happening through NIH and maybe through companies themselves. I don't think the companies have been terribly good about this. I've tried having them on a podcast I do, the Talking Biotech podcast, and they are not interested in coming on, even though we have a weekly subscription of 10,000 people that they could reach. I think they just want to play it close. at this point. There are some excellent people to follow. Dr. Paul Offit is the rotavirus vaccine creator. He's outstanding to follow, speaks very clearly about what this is and isn't. Dr. Peter Hotez is very good, and Dr. Michael Osterholm. Dr. Michael Osterholm has an outstanding podcast on, on this, and he's very good to follow as well and gives many good tips about communication.

This next question says, for the high pain responses, was this due partially because it's Because of the negative 100 degrees Fahrenheit in response by the muscle. So I think I can chime in on this one. I think number one, yeah, it's not injected that cold. It's brought to a normal temperature, or I'm not sure if it's body temperature, but it's definitely not that cold. And I think generally intramuscular vaccinations tend to, tend to be pretty painful as it is. As for the reasoning behind the elevated pain in this case, do you guys know why that might be?

Inflammation.

Yeah.

Yeah. So the antigen-presenting cells come in there and they trigger an inflammatory response. And the inflammatory response causes leakage locally in the area. And that increased fluids puts pressure on your nerves and that causes the pain. So it's a good thing, right? Because that means that, you know, for that leakage to occur means the inflammation has kicked in, means the antigen-presenting have gotten busy.

Yeah, that makes sense. Thank you so much, Asha.

You're welcome.

It looks like there's not any more questions, just lots of thank-yous.

So one thing I wanted to recommend also, if you're really interested, they're long podcasts, but This Week in Virology has— every week they have 2 or 3 podcasts, and one of them is by a doctor who's located in New York, and he gives, you know, he actually treats hundreds and hundreds of COVID-19 patients. And he sort of gives a clinical update every week on things that are going on and regarding the vaccine. You know, they're long podcasts, so they're not for the weak of heart. But if you really wanna get into the nuts and bolts of this, that would be the one to go.

Well, that's really great. Thank you very much to Dr. Bruininks for joining us today. It's always wonderful to see you. And very nice. Thank you very much, Ally, for joining us today and doing such a beautiful job with Talking about the New England Journal of Medicine paper.

Thank you.

And everybody out there, please go out and share, you know, share the, the link to this when we present it, and please go out and share this information. It's on all of us to share good science and the reason why it's important if we're going to get out of this faster.