- Hello, everyone! And welcome to Wednesday Nite @ the Lab, run 50 Wednesdays throughout the year, supported by the Wisconsin Alumni Association, along with partners UW-Extension, the Biotechnology Center, Wisconsin Public Television, and you.  I am Jordana Lenon.  I'm the Outreach Specialist for the Wisconsin National Primate Research Center and the Stem Cell and Regenerative Medicine Center, and standing in for Tom Zinnen, who's on vacation.  And I'm very happy to introduce our speaker tonight, Dr. David O'Connor.  Dr. O'Connor obtained a BS in Biology, Honors, from the University of Illinois at Urbana-Champaign and moved to UW-Madison, where he received his PhD in Medical Microbiology and Immunology.  He's now a professor in the Department of Pathology and Laboratory Medicine at UW-Madison, and is Associate Director of the Wisconsin National Primate Research Center.  His lab groups study the interplay between genetics, immunology, and infectious disease pathogens.  His research has contributed to our understanding of how HIV and AIDS undermine immune responses.  He recently developed the first non-human primate model for studying Zika virus infection during pregnancy.   He also serves on the National Institutes of Health's AIDS Research Advisory Committee, tasked with making recommendations about HIV/AIDS research priorities with the National Institutes of Allergy and Infectious Diseases. 

 So please let's give a warm welcome to Dr. David O'Connor.  [applause]  - Thank you, Jordana, and thank you, everyone, for taking the time to come and listen to a presentation about Zika virus tonight.  The first thing I want to say is that, in a sense, I am an accidental expert on Zika virus.  So, when I say that what I mean is that a year ago, about a year ago, this month, I was in Brazil with one of my colleagues on vacation, because my family has worked with our colleagues in Brazil on infectious disease issues for nearly 10 years.  And about a year ago we were looking for somewhere to go, and one of my friends from São Paulo said, "Why don't you come down and we'll spend a week at the beach and we'll have a good time?" And so we said, "Sure!" Unfortunately, the Brazilian economy is sort of in a tank right now, so that makes it actually very cheap to go there.  And I spent an entire week down there with people from Brazil, last year at this time, and the word Zika didn't get mentioned even once.  So what's happened is Zika virus has come on the scene very, very quickly.  And for reasons that I'll explain, it's a virus that we know nearly nothing about.  So because I was studying it just a couple of months before it hit the mainstream media here in the US, that made me the de facto expert because I knew just a little bit more about Zika virus than most other scientists and most other people who are studying viruses.  So I'll share with you some of the work that we've done at UW-Madison, as well as, hopefully, allay some of the fears that you might have about Zika virus. 

 And then after we're done, I'll be able to take any questions that you might have that I don't cover in my presentation.   So what is Zika virus? Zika virus is a virus that was first found in 1947.  And until very recently, this was one of the many, many viruses that scientists discover.  We're sort of like stamp collectors in this way.  We go out into the field; we take some animal or we take some person; we sequence all of the viruses that might be floating around in their blood and we catalog them; and we put them into families.  Really, just like stamp collectors.  And the vast, vast majority of these viruses, we then do nothing with.  The reason is there's not nearly enough research funding and not nearly enough research interest to justify doing intensive research on every virus that we possibly discover.  So typically what happens is someone discovers a virus and then does some basic research and asks: Okay, is this virus one that is likely to make people sick? Is it from a family of viruses where its genetic cousins make people sick? And then we do sort of this risk triage, this risk assessment, and we figure out whether a virus is likely to cause disease in humans or cause economically meaningful disease in livestock.  And then we decide whether it should be subject to more research. 

 And, in fact, that happened with Zika virus in the 1950s.   [David] This Internet connection is apparently not working.  [laughter] So, in the 1950s researchers did this sort of evaluation for Zika virus.  And what they discovered was that if they put the virus through enough generations of mice, they could get a virus that, you know, if coaxed just enough, could make mice sick.  But generally speaking, when they looked in people, they found that a lot of people had antibodies to Zika virus, but no one had recalled being sick.  So the conclusion was that Zika virus was a virus that was endemic in Africa, but wasn't capable of making people sick.   Now in terms of its genetics, Zika virus is most closely related to another type of virus that you might've heard of, called the dengue virus.  Dengue virus is a big deal, especially in tropical climates.  So it's thought that about 2 1/2 million people get dengue virus each year.  And I've never had dengue, to the best of my knowledge anyway, but I'm told it's really, really unpleasant. 

 And the thing that's most troublesome about dengue is that the first time you get it, it can be pretty bad.  It'll lay you up like a bad flu.  But if you get dengue two or more times, each subsequent time you get dengue virus, if you get a different type of dengue virus, you can be...  you can suffer from a hemorrhagic syndrome called "Dengue Hemorrhagic Syndrome" that can cause really serious, and in some cases fatal, complications.  So when we think about the viruses that are really closely related to Zika virus, we think about dengue virus.  And one of the things that I'll come back to later is this idea that Zika virus, because it's so closely related to dengue virus, may be functioning in some ways like a new type of dengue virus.  This is something that we have to bear in mind.  And one of the things, as researchers, that we did right away is we said: Okay, Zika virus is most similar to dengue virus.  So let's take the knowledge that we've applied for dengue virus and let's apply that to Zika virus and let's see if we can just generalize to Zika virus what we know about dengue.  And what I'll tell you in a little while is that there's a lot of things about Zika that are very similar to dengue virus, but there are also some critical differences that we have to be aware of when we're trying to figure out what we're going to do next with respect to Zika. 

 One of the things that's similar between dengue and Zika is that they're spread by the same types of mosquitoes.  So there's a type of mosquito you may have heard in the news: Aedes aegypti or  Aedes albopictus.  These are the same types of mosquitoes that spread dengue.  They also are capable of spreading Zika virus.  But one of the key differences is that we now know that Zika virus can also be spread sexually, whereas dengue is not spread sexually.  This is really a unique feature of Zika virus because it's really the only mosquito- borne virus that we know of that can also be spread through sexual transmission.   So what happens if people get Zika virus? The two main problems that have been associated with Zika virus infection over the last couple of years are what's called Guillain-Barré Syndrome.  This is a disease of the peripheral nervous system where people experience muscle weakness that eventually can lead to paralysis.  And this happens, we think, in about 1 in 4000 people who get Zika virus.  So not very common, but if you get it, it can be pretty devastating. 

 The bigger issue by far is this association between Zika virus and fetal abnormalities.  We now think that approximately 30% of babies who are born to mothers who  are infected with Zika in utero will go on to have some form of fetal abnormality.  One of the main research topics that's being pursued by our groups and lots of others around the world right now is trying to figure out exactly what abnormalities can be associated Zika virus; what percentage of pregnancies are going to be affected; and what is it that modulates that risk.  Is the risk greatest during the first trimester of pregnancy? Is it greater in women who have previously had dengue virus infection? There's a lot of unknowns right now that go into this percentage.  In fact, scientists who are doing studies in different countries are coming up with different percentages of babies who are born with fetal abnormalities, depending on how they define fetal abnormalities, exactly what goes into their definition.  And there also may just be differences from place to place in terms of how severe Zika virus infection is.   Nonetheless, for a virus that a year ago none of you in this room had probably ever have heard of, causing something on the order of 30% of pregnancies to have fetal abnormalities, is a big and scary deal.  So there is now a huge amount of research interest into the Zika virus because we need to understand how we can mitigate this risk, especially to pregnant women and women of childbearing age who might become pregnant.   So what sort of complications have been associated with Zika virus infection? Well, the one that's made the most news is microcephaly, which is a fancy way for saying babies who are born with unusually small heads.  And often times when you watch news reports, you'd think that microcephaly is the only consequence of Zika virus infection, but it's not. 

 Scientists have seen in babies who are born to mothers who had Zika virus unusual calcium deposits in the brain; excess fluid in brain cavities and in the areas surrounding the brain; missing or poorly formed brain structures; abnormal eye development; nerve damage; club foot; and unusually inflexible joints.  So there's really a spectrum of birth defects that have been associated with Zika virus infection during pregnancy.  And one of the great unknowns right now is whether babies who are born apparently normal to Zika-virus-infected mothers will eventually go on and have abnormalities that don't manifest until later in development, until the kids are one, two, five, ten years old.  So because the very first cohort of babies in Brazil born with microcephaly were only born last year, we really are going to have to follow these children for a number of years before we know what the full spectrum of possible complications will include.   So I like to contrast this in some ways to HIV/AIDS.  My background is as an HIV researcher, so I've been studying HIV for nearly 20 years.  And in the earliest days of the HIV epidemic in the early 1980s, what doctors saw were people who had end-stage AIDS: their immune systems had already collapsed, they were already being affected by these so-called opportunistic infections that people who have intact immune systems aren't susceptible to.   We then worked our way backwards and realized that people who were susceptible to opportunistic infections had likely been infected with the HIV virus, on average about 10 years prior.  And this passing from getting HIV to developing AIDS has been a major drag on our healthcare system because the cost of taking care of people with HIV has been so high.  It's estimated that the lifetime cost of care is on the order of about one-half million dollars per person. 

  So when we're talking about Zika virus, I want to emphasize that we're right at the very beginning right now.  So what we are seeing are babies who are being born with abnormalities now.  We don't know what other abnormalities might become manifest over the first few years of life.  But this is going to create a lifetime commitment to care.  And in the case of babies who are severely affected, economists have estimated that the lifetime cost of care might be more on the order of $10 million per infected person.   So when you hear news stories where the government is discussing whether it makes sense to spend $1 billion on a Zika virus response right now, it's important to bear in mind that every single excess fetal abnormality that arises due to Zika virus infection is probably going to incur costs on the order of $10 million.  So it doesn't really take very many $10 million-infections to get to the sort of billion-dollar price tag that we're currently debating in terms of whether it's worthwhile to have a really robust response to the Zika virus here in the US.   So, now I've talked a little bit about the problems that can occur with Zika virus and about how often they occur.  And so the big question that I want to know the answer to, and many of you probably want to know the answer to, is what happens next? Where do we go from here? First, I think it's instructive to look at where we have what's called active Zika virus transmission.  This means that the virus is going from mosquitoes into people, back into mosquitoes, back into people. 

 What you see here on this map in purple are countries where we have so-called active transmission.  What you can see is that, with the exception of Chile, United States, and Canada, Zika virus transmission is occurring throughout South and Central America as well as throughout the Caribbean, which is shown in the inset.  So the mosquitoes that can transmit the virus are transmitting the virus in all of these different countries shown in purple.  This is going to be a problem that's going to be with us for a little while.   Among the locations in the Caribbean that are experiencing a Zika virus outbreak, I think it does make sense to pay special attention to Puerto Rico because Puerto Rico is a US territory, and there is a major outbreak of Zika virus going on in Puerto Rico right now.  The way that they're assessing how bad the Zika virus outbreak is in Puerto Rico is by monitoring blood donors.  We knew that six months or so ago, if you were to sample 100 blood donors who came in to give blood in San Juan, none of them would have had Zika virus.  But over the course of the last couple of months, the incidence of Zika virus in blood donors in Puerto Rico has gone up and up and up.  And if you extrapolate through the rest of the summer, the people who work in the blood banks think that approximately 25% of all the people in Puerto Rico are likely going to be infected with Zika this summer, based on the prevalence and incidence of Zika virus among blood donors right now.   What is that going to mean for people living on the island? For the vast majority of people living on the island it's going to be nothing at all because, again, remember that Guillain-Barré is a very rare complication. 

 And so if you are an otherwise healthy adult male or female and you're not pregnant or planning on becoming pregnant, your risk of getting Zika virus is very, very low.  Ninety percent of people who get Zika virus don't even know that they've been infected.  And the remaining 10% might have a very mild itchy, inconvenient rash for about a week.  So if you get it and you don't have Guillain-Barré and you don't have the concerns of pregnancy, the fact that a large number of Puerto Rico are going to get infected this summer shouldn't be too scary.  In fact, it might actually be good news for reason that I will come back to a little bit later.  Nonetheless, with 32,000 or so pregnancies in Puerto Rico each year, this still leaves open the possibility that thousands of babies are going to be born with abnormalities either later this year or early in 2017.  In addition to the health costs associated with Zika virus, there are going to be secondary costs that impact tourism, blood transfusion, securing the safety of organ and tissue transplant programs.  And there's going to need to be an investment in mental and emotional health for women who are pregnant and don't have Zika virus, but have to carry this anxiety throughout their pregnancies.   Additionally, and it's an uncomfortable subject, but abortion politics, contraception politics are going to feature in the dialogue about what's going to happen in Puerto Rico with Zika virus later this year.  In fact, I got a little bit of a preview of that last week. 

 I was asked to speak before a Senate committee hearing on Zika virus and you could see very much the alignment of the different political parties with respect to how they viewed the most essential ways of combating Zika virus, both in Puerto Rico and if the threat reaches the US.  I think that some of these thorny issues are going to come to the forefront and with this coinciding with an election year, it's entirely possible that this is going to be a major driver of some news cycles between now and November.   Now what about in the United States? You've probably heard on the news people have talked about the Zika virus in the US.  And it's important to emphasize that right now...  well, actually these slides are as of about two weeks ago...  there are a bunch of what we call travel-associated cases in the US.  These are people who were traveling in Central or South America or the Caribbean, they came back, and they were diagnosed as having Zika virus upon their return.  None of these cases indicate transmission from mosquitoes in the United States, but among these 755 cases are more than 200 cases in pregnant women.  Now this is a bit of over-reporting because doctors are largely only sending at-risk women's samples off for definitive diagnosis because if you don't have Guillain-Barré and you're not pregnant or at risk of becoming pregnant, the fact that you come back with the Zika virus infection as a souvenir from your trip to Central or South America really isn't that big of a problem.  And so given the limited resources available for testing, people who come back and are just curious whether or not they have Zika aren't having their samples sent off. 

 So there is an over- representation of pregnant women in these statistics.  But in the territories, which include Puerto Rico, we have the converse.  There is a very small number of travel-associated cases, but a larger number of cases where there is local transmission from mosquitoes into people.  And again, a sizable fraction of these cases are in pregnant women.  This is a slide that I originally gave when I presented a version of this talk a couple of weeks ago.  I think it was two weeks ago yesterday.  So this morning I went back and I actually pulled the updated numbers.  And I'm going to show them here just to illustrate that this is a problem that continues to grow.  So, 755 became 934; 234 became 287.  The number of cases transmitted by mosquitoes has grown from 1,436 to 2,020. 

 The number of cases in pregnant women has grown to 250.  These stats are current as of about the last week of June.  So what this means is that as we get into the height of summer and the height of mosquito season, we are going to see a major surge in the number of Zika virus cases both in US states and in US territories.  The million-dollar question that I don't know the answer to and I don't think anyone knows the answer to is: How much mosquito spread can we anticipate happening in the continental United States? So the first question is: Do we have the mosquitoes? Answer to that is, unequivocally, yes.  As you see here on the left, we have the host range of the Aedes aegypti mosquito and on the right we have the Aedes albopictus mosquito.  What you can see is that these mosquitoes are widely distributed throughout the Southern United States, and they're particularly common in Florida and along the Gulf Coast.  Now if you look at the map, don't squint too much because if you squint hard enough you'll see that there is a little bit of an extension into the southern tip of Wisconsin.  But remember that when they do these sorts of surveys, they will take the identification of a single mosquito in a tire as evidence that a particular type of mosquito is in a state.  So we really don't have significant numbers of these mosquitoes in Wisconsin.  So we really don't have to worry very much about local transmission here in Wisconsin. 

 We do, however, need to worry quite a bit about it on the Gulf Coast in Florida and in Texas because there there are a lot of these mosquitoes.  And we also know from the dengue virus history that it is possible for these viruses to be spread in the continental US.   Now, one possibly reassuring feature of our history of studying dengue virus is that in recent times, dengue virus outbreaks in the US have been relatively small and self-contained.  There was an outbreak on the big Island of Hawaii last December, but really outbreaks of dengue are sporadic and typically brought under control through intensive mosquito-control measures.  That's probably because we do have better infrastructure than other places that are reporting significant mosquito-borne Zika virus transmission.  We have screens on many to most of our windows, we have access to air-conditioning.  We do have pretty good mosquito-control programs, though I'm learning more and more, that there's lots of room for improvement there.  There are some reasons to think that because of our infrastructure we are going to be better protected from local transmission of Zika virus than, say, Puerto Rico will be, but we really don't know.  We also don't know whether Zika virus is transmitted more efficiently than dengue virus by mosquitoes.  So we don't know how much we should be reassured by the fact that dengue hasn't spread very much in the continental US. 

 But clearly, if Zika virus does start spreading in the continental US, it will have similar secondary impacts not only on pregnant women who are going to have serious anxiety, but also tourism, blood supply security, organ transplant systems.  It really is going to take only a very small number of cases to have Zika punching above its weight with respect to how it's going to be perceived.  For those of you who remember the small number of Ebola cases two years ago, you can see and you can imagine how a very small number of Zika virus cases could lead to outsized public anxiety.   With that said, I think it's quite likely that there will be at least some local spread.  We don't know how much, as I said, but I think that we can expect that there's probably going to be at least some.  And as I said, even a small number of these cases can incur major costs.  This is going to be something that we're going to have to watch closely over the summer.   That's kind of a background on Zika virus and where we are as of right now.  So what I'm going to do is spend the next bit of my talk talking about what we've been doing at Wisconsin to study Zika virus and why researchers in Wisconsin, who are nowhere near these mosquitoes, have managed to be very involved in the public health response to Zika virus.   The story starts with me and some of my colleagues who have been studying HIV for a really long time. 

 And as part of our work on HIV we've been working internationally with partners in South America and Africa for a long time.  These are sustainable, long-term partnerships.  Here's a picture of me with my colleague Tom Friedrich, with our colleague, Denis Nansera, who's a pediatrician in Mbarara, Uganda, back in a hut in 2010.  We've been really involved in trying to take this technology that we discover in our labs here in Wisconsin and use it to help our colleagues who are studying HIV around the world.   In addition to our work in Africa, my longest-standing collaboration internationally is with this gentleman, in the middle, Esper Kallas.  Esper is a clinician researcher at the University of São Paulo in Brazil.  This picture was taken in 2005.  You can tell I'm younger because I had more hair.  [laughter] I'm shown here with my wife, who you can tell, is smiling because she's been married to me for 10 fewer years in this picture than she has been now.  [laughter] We have worked with Esper and his team on HIV, on dengue virus, and on hepatitis C virus projects going back to 2005. 

 We have exchanges between scientists from our labs.  So we go down there a couple times a year and show their scientists how to use the newest and latest and greatest tools that we developed in our lab.  They bring their scientists and clinicians up to Madison and receive on-site training.  And we learn a lot from them about how these infectious diseases are impacting their groups in Brazil.   In addition to our work in São Paulo, we also have collaborators in Rio de Janeiro.  Here once again is my wife, shown here with Renato Santana.  He is a professor from the Federal University of Rio de Janeiro.  Here he was actually giving a Skype lecture with my undergrad class on HIV.  We were in Florianópolis, Brazil, and he was talking about the Brazilian response to HIV to 100 or so undergraduate students who take a class that Tom and I run every fall.  And you can see our son looking bored, doing his reading in the corner. 

 This same day, as we were doing the Skype lecture, Renato came up to us and said, "You know what, guys? I need your help" because we have heard that there are a bunch of babies being born in the north of Brazil with really small heads.  And one of the things that Tom and I have done in collaboration with Tony Goldberg in the Vet School over a number of years is we've collected those postage stamps.  We've gone out into the African jungle, and we've sampled different types of monkeys, and we've looked for new viruses that haven't been discovered before.  So, Renato asked us if we could look in some of these babies to see if they had a new virus that was causing them to be born with really small heads.  And within a couple of weeks, it was pretty clear that Zika virus was the leading candidate for what was causing these babies to be born with small heads.  So we turned our attention from trying to figure out what virus was making the babies be born this way to trying to figure out how we could understand more about this virus.  Because as soon as we realized it was Zika virus, our next thought was: Oh, my gosh, we know nothing about Zika virus! And this isn't just us like, "I didn't know anything about Zika virus," but I didn't.  This was as a community.  If you looked in the databases that have papers, all the papers of all the scientific programs from around the world, there was something like 15 Zika virus papers over the last 50 years.  There really was not very much known at all about Zika virus. 

 However, it did make it pretty easy to get up to speed because you didn't have to read a whole lot.  [laughter] At the same time as we were learning about Zika virus from our colleagues in Brazil, Jorge Osorio, who's a professor in the Vet School, who also has had a long-standing research program in Colombia, where he is from, came back from Colombia and said, "Oh, my God, we're seeing cases of Zika virus in Colombia!" So Jorge's lab was actually the first to describe Zika virus in Colombia.  So early in November, Tom Friedrich, who had been talking to Jorge because he worked with him as a colleague in the Vet School and had been talking to me because we were talking about Zika in Brazil, put this together and said, "You guys really need to talk to each other." So we did and we decided that one of the things that was going to be really important in learning about Zika virus was trying to develop animal models that we could use to ask very focused questions about Zika virus very rapidly so that we could inform the sorts of studies that were going to be done in people, but that are going to require very large cohorts and are going to play out over a much larger period of time.   After Jorge and I decided that we were going to start working on this together, we realized that we were missing some critical buy-in.  So fortunately, I was able to convince Jon Levine, who's the Director of the Wisconsin Primate Center and is shown in the center here, and Buddy Capuano, who's the Attending Veterinarian and the Associate Director for Animal Services at the Primate Center, that we really needed to start thinking about doing Zika virus studies with experimentally-infected macaque monkeys, where we would give macaque monkeys a defined dose and strain of Zika virus, and then follow what happens to those animals throughout infection.  Fortunately, they both immediately understood the potential implications of this and offered the full support of the Primate Center.   And, finally, this was sort of an embarrassing admission that we all had.  We realized that if we're studying a virus during pregnancy, we need to know something about pregnancy.  And short of the fact that I have a son, that was about the extent of my knowledge of reproductive biology and obstetrics.¶¶¶¶¶¶[laughter] So we needed to get specific expertise that could help understand the many dimensions of studying an infectious disease that infects pregnant women  and infects the fetus in utero.  So Ted Golos, who has been studying reproductive biology, reproductive health, and infectious diseases during pregnancy for decades at the WNPRC, joined our team in early December as the obstetrics component. 

  And so we really have a multi-disciplinary team that spans all of campus working on this.  And over the course of the next couple of months, our team has grown to encompass even more dimensions.  Here is Dawn Dudley, she's on the right, she's a senior scientist in my lab.  Here we are down in Rio de Janeiro trying to help them process some of the first Zika virus samples that they got in their lab back in February to try to characterize what sort of tissues have Zika virus in them.   Sallie Permar is a colleague of mine from the HIV community.  She's at Duke University.  She contacted me in late December and said, "I've been studying congenital cytomegalovirus infections in women for the last several years.  Have you heard about this Zika thing?" And so she's been part of our team, bringing more expertise studying congenital infections that can affect fetuses.   Jorge has an extended team shown here down in Colombia studying Zika virus and other mosquito-borne viruses.  And as time's gone on, one of the things that's just been really awesome has been how much expertise we have on this campus that I had no idea about, to be honest. 

 It's kind of embarrassing because I've been here since 1997.  We have people like Christina Newman, whose job for a long time was to go set mosquito traps down in Chicago to capture mosquitoes.  She'd bring them up here to try to grow them in the insectary.  But mosquitoes don't like growing inside-- outside of their natural environment-- so what she would do, because they wouldn't eat the synthetic food that she gave them, she'd give her arm out and they would feed on her arm.  They'd take a blood meal.  So they established the insectary using mosquitoes that were feeding on Christina.  But she has become one of our local mosquito experts.   And we've also drawn on a lot of the expertise that exists in the medical school.  One of the things that we want to do during pregnancy is look at how animals develop during pregnancy.  And to do that the most sensitive technique is MRI. 

 Here you have Kevin Johnson, who is a medical physicist, whom we recruited along with several maternal fetal specialists, to help us look at the MRIs of developing fetuses.  Again, we really were shameless in going out throughout campus and getting expertise from anywhere we could find it to help us answer some of these key questions.   I think there might be only one more of these.  These are two neonatal specialists who spend most of their time looking at ultrasounds from human babies.  But they got trained-- They went through all the rigmarole necessary so that they could come back and also use their expertise to look at monkey babies and try to assess whether there were any developmental issues in the monkeys.  It really has taken a huge team to get where we are right now.   What we've been able to do as part of this team is develop the first monkey model for understanding Zika virus.  In summary, what we found is that macaque monkeys, which are the kind of monkeys that get used in most biomedical research, can be infected with Zika virus and exhibit similar symptoms to people who are infected with Zika virus.  That is to say most of them don't get sick at all.  But they have a very similar trajectory of viral response and immune response as has been observed in people. 

 We see that the virus is detected in the same fluids for about the same amount of time and, critically, macaques have been used to study reproductive biology for decades.  So unlike other animal models like mice and rodents and guinea pigs, macaque pregnancies are generally very similar to human pregnancies.  They can be broadly subdivided into three trimesters; each trimester has key developmental milestones that track very closely with those in humans.  So this actually makes it a very good model for trying to understand how a virus like Zika could cause fetal abnormalities.   I'm just going to show you a couple pieces of data from our studies.  I have one piece of really good news and one piece of not-so-good news.  Let's start with the good news first.  The good news is that all the data that our group is generating and that others are generating points to the fact that a Zika vaccine should be successful.  So as someone who studies HIV and has studied it for a long time, I'm very gun-shy about saying things like, "a vaccine should be successful" because, of course, we said that about HIV back in 1984 and here we are 35 years later and we still don't have an HIV vaccine.  But there's good reasons to think that making a vaccine for Zika should be conceptually easier than making a vaccine for HIV or other more thorny viruses. 

 The reason we think that is because we infected a number of- and these are non-pregnant animals-- we infected these animals with Zika virus.  And what we did is we measured the amount of virus in the blood.  What you can see is that within one day of infection there was some Zika virus in the blood, but that by about a week after infection the animals had pretty much controlled their infections.  They had cleared their virus, there wasn't any virus left.  There were occasional very low-level blips out to about 20 days, but, for all intents and purposes, the animals had cleared their virus within a week to 10 days.   We then waited another about eight weeks, so 70 days total, or 10 weeks from the first infection.  Then we gave them the same virus again.  The question we were trying to test is whether the immune response that was elicited by this initial infection, when the virus was replicating, would be sufficient to protect the animals from being re-infected if they were exposed to the same virus again.  And as you can see from the arrow where it says Rechallenge, we saw no evidence of any virus replication after the animals were re-challenged with the same virus.  So this suggests that the antibody response and that the immune response that got made against the first virus protects them from re-challenge. 

 Let's think about the implications are for this in a place like Puerto Rico.  It means that if you're a woman of childbearing age who's not pregnant right now, arguably the best thing that could happen for you would be to get Zika virus, develop protective immunity, and then if you get pregnant, you should be at significantly reduced, if not zero risk, for having Zika virus-related complications during pregnancy.   It also means that if you are scared about Zika virus, the best thing that could happen would be for there to be a wave of Zika virus that comes through a community, leaves the entire community immune, such that the mosquitoes won't have vectors that they can use to continuously transmit the virus back and forth between people and more mosquitoes.  Indeed, in some of the outbreaks that we've been going back and looking at that happened in French Polynesia and other parts of the Pacific in the last 10 years, that's what we've seen.  We've seen huge numbers of people being infected in places like Yap Island, and then the virus goes away and isn't heard from again.  Again, this is consistent with the idea that people have immunity.  We don't know how long it lasts, but that they have immunity that protects them from being re-infected.  That's the good news.   The bad news is that some of our data also suggests that the risk to pregnant women might be even higher than was previously appreciated.  Our reason for thinking that is because, in contrast to non-pregnant animals, whose amount of virus in the blood is shown here again. 

 So, this is the same data that you saw in the last slide.  When we look at two animals that were infected with Zika virus in the first trimester of pregnancy and two animals that were infected in the third trimester of pregnancy, we saw that in three of the four animals there was a dramatically-prolonged detection of virus in the blood, relative to what we see in non-pregnant animals.  So this suggests that the risk to the fetus might be much higher than currently thought, in part, because the virus sticks around a lot longer.  That provides a lot longer time for the virus to infect the fetus.  Now we do not know why the virus persists for so long-- yet.  But one of our leading hypotheses is that the situation is actually even a bit more dire than that, and that what we're actually seeing here in red are fetuses that are infected with Zika virus.  The mom infects the fetus.  The mom clears her infection because she has good immune responses, just like we see in the other animals, but the fetus doesn't.  What the fetus is doing is shedding virus back into the mom.  What we are detecting here is fetal virus that's detectable because the fetus and the mom are sharing the blood stream. 

 If this is true, it suggests that you have a duration of infection in the fetus that can vary from about a month to two months and possibly be associated with adverse outcomes.   What we're trying to do right now is to figure out what the impact of these virus infections is going to be on the newborn brains and other tissues.  So this is always the part of the talk where I have to say, "It's to be continued." Because we're just now at the point where we're delivering these babies by C-section and then we're dissecting them to look comprehensively at whether or not there are any abnormalities in the newborn, and, if so, where they're located.  What tissues and organ systems are affected? How were they affected? And, unfortunately, for the purposes of presentation anyway, it's about a 2 to 3 week process to get the tissues stabilized and prepared for analysis.  We're still in the midst of that 2 to 3 week process right now.  So I don't have any information on whether the newborn babies born to mothers who had this prolonged viremia have any adverse outcomes yet.  But we should be getting that data over the next 4 to 6 weeks.   We're also very interested in this idea that there might be interactions between dengue virus and Zika virus because the viruses are in the same place.  Some people have speculated that if you have previously had a dengue virus infection, you might be at a higher risk of having a severe Zika virus infection.  This is something that we can model in animals much better than you can study it in people. 

 People don't know what their dengue virus history was.  They don't know when they were infected, they don't know how many times they've been infected.  We can control that precisely by having animals that are challenged with dengue virus and then go on to receive Zika virus to see if Zika virus is worse than in animals that haven't had dengue before.  Additionally, just in the last couple of years, the very first dengue vaccines have been licensed.  So Mexico has licensed the vaccine, other countries have licensed the vaccine, and there are advanced clinical trials going on in a number of countries right now.  If natural dengue virus infection does change the risk of having adverse outcomes following Zika virus, what about dengue vaccines? If we vaccinate an entire population against dengue virus, are we going to modulate in some way what their future susceptibility to Zika virus is? Again, for the vast majority of the population, I think I'd go ahead and vaccinate for dengue.  But this is going to be something that we're going to have to think about when it comes to vaccinating women who could be potentially at risk of Zika virus in the future.  It also gives currency to the idea that we should really be focusing on trying to develop combined Zika and dengue vaccines.  But developing vaccines that target all the dengue subtypes and Zika at the same time, is a much bigger ask than making a vaccine just against Zika or just against one of the dengue serotypes.  The third thing that we're looking at right now is whether or not the virus persists in a replication-competent form for longer periods of time in tissues than it does in the blood. 

 I told you that within about a week to 10 days, the virus is pretty much gone from the blood.   But almost by accident, we looked at one of the mothers who had an enlarged lymph node at the time the baby was delivered by C-section, and we found that, much to our surprise, there was still detectable viral nucleic acid in the lymph node about two months after the animal have been infected.  Now, we don't know whether that virus is replication-competent or represents sort of a viral fossil of an earlier infection.  You can imagine that especially for people who are trying to secure the tissue and organ supply, it's vital to know whether or not tissues and organs that are going to be transplanted could be harboring the Zika virus, because while the risk to the general population is low, the same might not be true for people who are immuno-suppressed because they're receiving immunosuppressant drugs to tolerize them to tissue or organ transplant.  So it's really important to figure out whether this virus that we're detecting is actually capable of replicating and being infectious.   So...  The last thing I want to talk about is something that I'm particularly proud of because it represented a bit of a change in how we do science and it really required the buy-in from our entire team to do this.  This is the idea of real-time data sharing.   This has been a concept that's been kicked around in science for the last several years, in light of some of the last major infectious disease outbreaks.  It began right around the SARS outbreak in 2000, 2001. 

 In 2009 when we had the H1N1 pandemic flu, a lot of people were talking about the need to share data more quickly.  And then some of the investigators who were studying West African Ebola really put this into action and made a lot of their data available in close to real-time through public repositories so others could go in and analyze it.  In fact, one of the things my lab did was we took some data from West African Ebola patients, and we asked a question that was totally different from the questions that the study authors had asked.  And we eventually got into contact with them and put together our own manuscript that basically looked at whether or not someone who is co-infected with the second virus had reduced susceptibility to Ebola.  That was really an eye-opening experience for me because it showed that the data that we're collecting and the data that we're generating, other people might want to use in ways that we could never imagine.  Other funding and people who pay for the research and the journals that publish the research all fortuitously agreed, in principle, to this statement on data sharing and public health emergencies back in February.  The idea was that we should try to make Zika virus research as available as possible, as quickly as possible.  This goes to one of the real problems with scientific knowledge communication in general, which is the long time from when data is collected until it's made available to the public and the rest of the scientific community through published papers.  So the general process for doing a study was kind of like this: you start the study; you do some experiments; you finish collecting the data; you analyze the data; you write up a manuscript; you submitted to a journal; the journal submits it to a series of peer reviewers who rake you over the coals and tell you your work is terrible, but that they are willing to publish it if you make the following 36 suggestions; you respond to these individual comments one at a time; Hopefully, eventually, the reviewers are satisfied and the manuscript appears as a published paper.   In the case of our initial Zika virus model work, that timeline looked like this. 

 And this is actually a very, very fast timeline, as research goes.  We started our study on February 15.  By March 15, we had collected data from the first four weeks of infection.  We wrote the paper over a process of about two weeks and submitted it to a journal.  We responded to comments, once we got them back in the middle of May.  Then, eventually, the paper was finally published last week.  You're looking here at a time window of more than four months between the time when we started the study and the time that we ended up publishing it.   For a virus whose trajectory is changing as quickly as Zika, that has such public health importance, this is simply too long.  We can't afford to let data sit siloed in individual labs and at journal editors while people are continuing to get infected, especially when we have the greatest opportunity to intervene and try to mitigate the threat.   I would argue that it's actually not particularly ethical to delay the distribution of research results for months during a public health emergency. 

 We asked: What could we do differently? One thing that people in science have been doing is they've been publishing to what are called these "preprint servers." So at the same time that they submit a manuscript for publication in a conventional journal, they put a PDF of it online in these indexed preprint servers.  This is good because it saves considerable amount of time.  We posted our data in a preprint server the same day that we submitted it, which was March 31.  But this was still quite a while after we started the study, so we took it one step further.  What we did is we decided to put all of our data online in real-time.  This takes advantage of the fact that my lab actually has pretty good computing expertise and bioinformatics expertise.  This model wouldn't necessarily work for everyone, but we were in a position where we were already going to be storing our data and making it available to the really large team of UW investigators I talked about earlier.  So we decided, well, rather than restrict it by password or user ID to the people who would want to see it here at UW, we'll just open it up to the public and see what happens.  It was quite the experiment.  But it turns out that it's been really quite successful. 

 So all the data is available, and information on all of our participants is online, along with how we've done the studies, the raw data that people can go in and re-analyze, as well as annotated data.  One of the things that we learned pretty quickly is that a lot of people who are interested in this data didn't really have the stomach to go and analyze it themselves.  We took to writing brief summaries that kind of guide people through what the data means.  One of the people in my lab, who actually just returned from giving a-- to being a social media outreach expert at the Juno orbiter this weekend took over the lab's Twitter account when she realized I had no idea how to really use Twitter-- which is absolutely true.  She started putting some of that information on Twitter and we actually started getting a lot of feedback.  One of the really neat things that's happened is other people have interacted with us about our studies in real-time.  In some cases they've taken what we've put up online and given us suggestions about how we can do things better.  So in the top example here, we had tweeted that there was some data that suggested that a number of the viruses that were called one thing, one of them wasn't like the others.  And this researcher from the University of Birmingham in England immediately wrote back within a few hours and said, "Yes, we also noticed that that sequence was wrong, "but we actually have already taken it one step further, "and shown that it was a mosquito virus "that was incorrectly labeled in the database of viruses, not an actual Zika virus from a primate or a person."  On the bottom, you see an example where a woman who has led studies in human pregnancies affected by Zika, who's also seen this idea of extended viremia, or prolonged detection of virus in the blood, saw some of our data and suggested that we do some additional experiments that she wanted to do in people, but she wasn't able to do.  This has turned out to be really good, and if you're curious at looking, the URL is there zika.labkey.com I'm just going to close by saying that I think, in a way, this is what Wisconsin and the University of Wisconsin is all about. 

 Because if you think of what the Wisconsin Idea is, it's the idea that we take knowledge that's generated at the University and share it with the community for the greater good of all and really have knowledge that transcends the boundaries of the classroom, or in this case, the research lab.  So it's been really great to have the University of Wisconsin be at the forefront of this type of data sharing and have a team of people who are so willing to try something that's pretty far outside the orthodoxy for scientific research.  With that, I'm going to close by acknowledging just a small number of the people who have helped with this.  In blue, you have the people who have been the project leaders in various parts of this: Dawn Dudley.  Emma Mohr is a pediatrician who is a Fellow in my lab and has brought a valuable clinical perspective.  Tom Friedrich, who I've worked with for nearly 20 years on HIV.  And we're somewhat inseparable when it comes to doing this sort of work together.  Jorge Osorio and Matt, who brought the mosquito virus expertise.  My wife and colleague, Shelby, who did some of the virus sequencing in our projects.  Sallie Permar from Duke and her team. 

 Buddy Capuano, the attending vet at the Primate Center.  Ted Golos, who led the obstetrics.  NIAID, which is a part of the National Institutes of Health, that when they learned that we were about ready to start the Zika projects were able to use some unconventional funding mechanisms to provide support for these studies, as well as the Wisconsin National Primate Research Center who galvanized the studies by providing seed grant support.  And then, our colleagues in Brazil: Amilcar Tanari and Renato Santana at the University of Rio de Janeiro and Esper Kallas and his team at the University of São Paulo.  So with that, I will stop and take any questions.  Thank you.  [applause]