Peered. As I understand it. It was effort that was spearhead spearheaded particularly back in the Clinton administration with participation by some of the speakers today, Lake Harold Varmus and Anthony Fauci in 19 the. And giving rise then four years later to the actual in vaccine development. They are true expert on the subject, vaccine development has been a central driver of the progress made over the past couple of centuries because after all Edward Genres coining ever the term vaccine and the application of that to a small boy with the Cowpox vaccine started this enterprise, almost would turn to different animal model. Inoculateing rabbits with rabies. Like pasture tried out his vaccine on a young boy I hesitate to think what the Irb Irbs would have to say about that. But in both cases, despite imperical production methods, the boys lived and vaccines and others developed afterwards which are truly one of the great triumphs of medicine. Transformed humanity's inter- inter-actions with disease and saved countless lives. Here at NIH we've been working workingworking on vaccines from the earliest days of our creation as small hygiene I can laboratory over the years. In clueding that for polio. So, in the early 19s an effort began here led by Tony Fauci a century after the rabies experiments to try to work on disease that was arguably as frightening as rabies had been and threatening to become more pervasive. Of course I'm speaking of HIV/AIDS. And the search for that gave rise to the Vrc that was here to celebrate. With at times frustrating some will be talked about today in ways that can give us all hope. That ultimately this goal will be achieved. I want my colleagues fill out the rest of the historic center, story for the VRC but I would like to pay tribute to the center itself as you came in you may have seen photographs of the many people who work within this center, a remarkably hard work working dedicated working with other intramural investigators and the extramural research community to translate basic science in vaccines that the VRC is indeed a model of that effort we are certainly studying its progress closely as we work to design NIH's proposed center. Plague humanity. It's they're to say we've come a long way. We've moved in to the Era of partnerships and approaches that are now available, something that could not have been imagined by Genre or Pas teur the promise is tremendous. I'm glad to say in conclusion that this is an effort that is conducted with a very clear eye towards our world as contributor to mobile health, observe infectious diseases no no country boundaries, it was Paste Pasteurur says knows no country because knowledge belongs to humanity. Lights of science in these darker corners of infectious diseases that are untolled humor suffering. Doing the VRC is a mission that fits nice plea and beautifully with the overall NIH aiming to improve the health of the people all around the world. Happy anniversary. VRC may you have many more it is now my privilege to introduce the next person to make in freethrow duck tore reremarks none other than Dr. Harold Varmus once the director of NIH now the director of the national cancer institute. Harold. >> Some initiative that was undertaken with incredible speed speed, minimal bureaucracy a summary of the events that led to the creation of the center that we're celebrating today. There was still considerable skepticism about the progress being made against AIDS and it was recommended by one of my colleagues, actually Phil sharp that we undertake a large study of AIDS research funded by the NIH. And agreed to chair the committee of distinguished scientists who looked at all aspects of HIV related research and among the recommendations they came up with was one to pay greater attention to immune responses to HIV to try to learn more about human immunology, to give greater attention to the question of why we've been un unable to make a vaccine, in particular they suggest that we look more carefully at the failures than we had been doing. And all of that was part of the information that bill Paul, who was then the first -- was the director of the office of AIDS research chewed on for some time to try to figure out how to respond to aspects of the la veen report. Using the building point to try to make a better contribution to worldwide efforts to develop HIV vaccine. And we discussed this intramural intramurally, we had a number of groups gathering for research that was many Lance on the campus that were interested in the vaccine. Or developing a vaccine, we had support from the national cancer institute and from NIAID trying to do something to develop a more active center. And then in the I think it was March of 1997 Tony and I and few others were asked by president Clinton to come and give a brief briefing on the current status of research on HIV and AIDS and during the course of fairly long briefing, which was bill Clinton listened very attentively said very little because he had severe laryngitis we brought up the question of whether we should develop a center on the NIH campus for developing vaccines he is special plea a vaccine against HIV. And we didn't say much during the meeting we had a little reception afterwards in the visiting line he whispered to me that he was very taken with the idea would like to follow up with it. And this was not an Idle gesture a few weeks later I was called with you a speech writer who was preparing a talk to be given at the commencement of Morgan state university, the president would like to say something about a commitment he was going to make to develop a vaccine center on the NIH campus. And that speech was made, it was a commitment very late stage in budgetary year to build a center that would be physical as well as" electric towel, in that year year's appropriation there was allocation, times were better than now from the point of the fiscal state of the nation that included in that budget bill for Fy-the 8 was the allocation to begin construction of the vaccine center. And indeed a year later the fall of 1999 the cornerstone was laid , Dale and Betty Bumpers who were long term advocates for childhood vaccination programs moving Arkansas from the bottom of the list to the top of the list helping to ensure that almost all children in this country received vaccination were on hand, along with bill Clinton and secretary Shalala and many others and within another two years, the building was up. And happily during that time we had remarkably successful search for a director of the center, who remains to minute in charge of the center, Gary Nabel, that search was significant not only because it brought Gary to the campus but it brought Betsy Nabe Nabel as well who became most of you know the director of the blood institute. We brought toga distinguished team the first man necessary takes of the team effort that would characterize the remarkable center that Gary has developed intellectually to correspond to the wonderful physical facility that you see before you. From the beginning, we had decided not to call this an AIDS or HIV vaccine center, but instead a vaccine center with the supposition that vac is in inology and I am mu that logical basis of making vaccines was permissible. In fact and abandoned the search for HIV vaccine which in case it would still be very helpful to have a generic vaccine center rather than one that was dedicated to the needs that was no longer expand. Happily although we don't yet have an effective HIV rack seen the center has devoted attention to many other topics, most prominently e-boli and importantly over the last couple of years especially influenza vaccines, recently Gary and Tony and I sponsored a workshop to think about developing an EBB vaccine another way to substantiate the idea that the center would be one of interest in promoting global health because an EBB vaccine would be helpful not only in protecting against infectious monobut also way of Poe senescence shallly various neoplastic diseases that EBV contributes to. Finally I'd like to point out one additional aspect of the center which I don't think will be addressed all that aggressive aggressively in today's symposium but it's important to me, that is that by creating a center for vaccine science an era where segments of the public seem oblivious to the well benefits of vac say sakes, the center sends a signal which is occasionally articulate in print as well. About the importance of maintain maintaining a strong attitude toward development of new vaccines in that era from some argue abandoned vaccines because of poorly substantiated or even salacious claims of their toxic effects. I'm very pleased to be able to point to the vaccine center as a place where great science is done to represent the idea that vaccines are one of the most powerful ways to develop here at the NIH and many other places to protect human beings against the ravages of infectious disease. So, thanks very much for the speakers for coming today I'm looking forward to being stimulated by the vaccine center this afternoon as I have been in the past. Thank you. [APPLAUSE] >> Thank you very much, Francis, Harold, for knows nice in freethrow duksz. I'm going to lead off the discussion part of this by filling in a little bit of the everything he said was correct which was one minor thing which I'll bring up in a second. And then take about a 30,000 foot view of some of the details you are going to hear from Gary when he talks about the accomplishments of the center. First of all, the history that Harold gave you was really quite correct and as you can see in my first slide I really want to recognize that although the responsibility for the AIDS research agenda fundamentally fell in to my lap as director of NIAID if it had been for Harold and bill Paul persistence and insight in to realizing the importance of the immune response and need for vaccine I I -- we have the vaccine center not in the form that it is now. I want to make sure that all of you who may not be familiar with the history realize the papers of that. Now, the story about the White House is entirely true except the one thing that Harold got wrong was December the 3rd, 199 6. I want to point that ought because Harold makes one Mis Misspeak every 20 years which means that anything he says for the next 20 years you better believe because it's going to be true. Here is the screen in the White House. We were invited by the president and there's Harold actually there and Bill and I and group of other people and Harold and the president really wanted to hear about the science. He's right, he was speaking like that during the discussion and when he heard I was the director of the national institute of allergy and infectious diseases he immediately wanted fob referred to an allergist. But which I'm not. Some of you who are familiar with this would notice, he is tellly -- really -- he was very interested in the science so I decided I would talk to him about the co-receptors of HIV, the CCR5. I tell the joke, notice the expression on Al Gore's face. He was imagining that it was not ed or Rick and colleagues who discovered it receptors that it was his, no, Mr. Vice president, that's the Internet that you discovered. Oh, yeah, the Internet. But in any event Harold was quite right as we were walking out it was actually out in to the rose garden for a press conference the president stopped us, Bill and I and Harold and asked Harold exactly the same question, what could we do. And what happened, he actually turned to Leon Panetta the chief of staff at the time and whisper whispered something to him. Which had us all get real flush of a good feeling because whenever a president tells something to the chief of staff, you know it's going to happen. That was really history of how it happened. And Harold is quite correct, things moved extraordinarily rapidly even within the framework of how things move in the government. This is the president at that commencement address at Morgan state that they asked Harold to make some suggestions for, this is what actually Harold told him that would be a good idea to do. He said today I'm pleaseed to announce the national institute of establish new HIV vaccine research center dedicated to this crusade. And in fact things did happen, it went up really quick plea, here is a picture of Donna Shalala that was secretary at the time with the bumpers here dedicating the plaque which stands now at the physical structure, that was on June 99 of 1999. And things moved quickly, we needed a leader, someone with an extraordinary intellect who gets things done. This is what the vaccine center looked like, I don't want to go in to any of those details now, you are going to hear about it from Gary. But we were very fortunate in getting Gary to not only lead in the design of it but also to have the foresight for the need for the types of vaccine development and valuation facilities that are important part of this entire enterprise. The building itself is the GMP production, pilot plant in Frederick. The clinical trial operates out of the NIH clinical center and the immune assessment laboratory in Gaithersburg. We also have a very interesting mobile unit which actually takes the research and not only the research but the education message out in to the street, because Washington D.C. far those of you not from this area as you know is really equivalent in so many respects to sub subsaharan African country when it comes to the type of infection and timing of infection that we see. I make rounds every week I'm absolutely amazed that the patients that we get from the Washington D.C. district it is not at all unusual for them to present to us with a CD4 count of ten and viral load of several hundred thousand even here right in the nation's capital. So, these units which go out there and get people vaccinated in our vaccine trials that we have, mostly with Bonnie -- Barney Graham and his colleagues colleagues, this is a picture of Gary when we hired him it was extensive national search, I don't need to tell you how lucky we are to have had Gary to accept. It's unusual to get a superstar as a director, it's open more un unusual to get a superstar as a deputy director. So we are proud that we have both Gary and John here leading this. But the real guts of the program program, in addition to Gary and here they are looking like they don't generally look during the day because they got dressed up for this, actually, most of them get dressed up, some get dressed down for pictures. He took his tie off. What's happened over the subsequent years. We have over 200 employees, the numbers of publications which we department like to just count publications, the publications themselves are numerous but they're impacting. We have vaccine products that have been used in clinical trials, we have a number of clinical trials and the critical issue that both Harold and Bill and I and others insisted upon was in one physical place to have what we call the soup to nuts phenomena where we actually use fundamental development of concepts at the very basic level level. And have the capability of taking that all the way to clinical trials. That really in many respects is extremely unique, but as we know now ten years later highly, highly effective. Now this is a slide that I show show -- I used to show it every year to the congress but they don't even give us an opportunity to get in front much the congress, any more, that's the truth, sad as it may be. That we used to talk about and still do, but we don't do it face to face except in private meetings about the importance much emerging infectious diseases. I originally drew this slide myself out of a national geographic map, because I wanted to put HIV as an emerging disease together with others, and every year I would show it at the congressional hearings add one, two, maybe three emerging infectious diseases. Many of which are amenable to vaccination. So the first one was HIV. So when you have disease like HIV, you know we're approaching the 30th anniversary of the recognition of the first cases reported in MMWR on June 5th of 1981. That we now have at least 33 million people who are living with HIV and this is the critical number, 2.6 million people who get newly infected each year. This is, I think, I've been thinking about that I mentioned it to Peggy Johnson as we were getting ready to walk in here, is that this slide itself is the most compelling argument for the need of a vaccine. Because we have here in the United States 56,000 new infections, and it's been flat for at least ten and probably closer to 15 years. That's in the developed world that has the exposure and capability of health care delivery tactics and all kinds of things that we do to get people to avoid the risk. And yet we've been unable to get below that. So even though we have what everybody is talking about for those of you not in the HIV field, the big thing now, everybody talks about is combination HIV prevention. Appropriately so. But paramount of all in combination is going to be an HIV vaccine. So whenever we talk about what we need in development of things that come from concept, we talk about scientific opportunity and public health need. And right now in 20111 I don't need to convince anybody that we have a public health need but the scientific opportunity is really getting very exciting. Now, many of you familiar that the first after 23 years of try trying, the first modest success of an HIV vaccine was the RV144 otherwise known as the Thai trial. Proofed that we could do it. What is going on right now and this is being spearheaded along with other good laboratories throughout the country, is what we call structure-based HIV vaccine design. This is very well described in a number of papers, I'm not going in to the details of this types of scientific finding, I want to give you more of a 30,000 foot look over few minutes of time. That is to take a look at what the structureally defined conserveed neutralizeing epitopes are and there are a few not the least important of which is the CD4 binding site. Again the VRC with Peter and his colleagues and John Mascola all the group that I showed you was involved in. This is adaptive slide from review by Dennis Burton in " "Nature immunology" that I adopted by adding some of the antibiotics in which he was describing his PG the and PG6 team. One of the most important of these antibiotics that da D.A. antibodies that are leading the way or the VCR01, 2 and 3. And in fact in series of rather classic papers that have evolved from the VRC we know now that you can identify these neutralizeing epitopes by what we call the combination of binding and structural definition of what those epitopes are. And it is very clear to make a long story short that it's look at the diverse HIV strains that are available that VRC01 and 2 monoclonal antibodies neutralize more than 90% of the globally diverse strain. In addition to that there have been a number of important concepts of how you can manipulate at the molecular level what surrounds these epitopes in a process called re resurfacing of stabilized course course, which conserve not only only -- which can deserve as probe for the antibodies but as template for immunogens, because it is difficult at best to identify the epitopes, it's even more difficult to put them in immunogenic form. In that regard, the VRC again among other labs as leading the way to try and develop an immunogenic capability for these epitopes by things like specific antibodies using epitopes scaffold as shown in this paper by Peter and his colleagues. This is a quick buzz through some of the things that are going on in the VRC. But as Harold so appropriate plea said when we sat around the table in his office with Bill and with a bunch much other people, we were very clear that we didn't want to make this just the HIV vaccine research center. But make it a vaccine research center that could address many of the problems, even though it was geared heavily towards HIV. One of those is the e-boli virus which appears on that slide that I showed to congress, again, with Nancy Sullivan and Gary and his Sol lesion have really spearheaded the way, both in proof of concept for a prime boost in primates to series of papers that made the transition from nonhuman primates to human studies. SARS, I think, even though SARS is no longer on people's radar screen, what happened with SARS I think manifested the best of the VRC. Let me explain to you why we say that. You all are familiar with SARS, it cropped up ununexpect edly as a true emerging infection, we describe theta's never before recognized in the human civilization, re-emergeing like west Nile some a different store reeks, it's been around awhile we get it in the United States probably by a fluke of transportation. SARS hit us hard both health wise and economically. In 2003 as shown by the more than,000,000 cases and close to 8 00 deaths. We didn't know it was going to burn itself out. I mean, it's interesting when you are living through history you don't know what the end became is going to be. So it was very clear that we needed a vaccine. So the VRC proof of concept for a SARS DNA vaccine was first done in mice and then in a phase one clinical trial and this timeline is very interesting. I put this slide in because it is the truth that this is truly the world's indoor record from the time a microbe or virus was first identified in March of 2003 to the time it was entered in to a clinical trial in December of 2004. Probably the most experienced of the vac sinologies, if that ain't light speed then nothing is light speed. Same holds true for west Nile virus. Which again, the group has been major leaders in the proof of concept for a human DNA vaccine that you can see by the titles of these papers and dates in 200 2007 then most recently again in 2011. Chicken virus, I remember one thing that was always get a laugh in medical school when you were studying chicken because it sounded so funny, it was -- Chik Chikungunya it's a very serious disease in the developing world. And then only as is often the case with diseases that afflict those -- everybody got excited when it was seen in northern Italy and again, the VRC sprung to action with the development of a Chikungunya virus what was shown to be protective in nonhuman like ply mate. Another very important triumph for the VRC. Then as Harold mentioned influenza. Interesting disease for those of you who don't follow it carefully because it fulfills our definition both of re-emerge re-emerging infectious shun because it comes back in different form every season as well as brand new virus as it done within you get a pandemic as we saw in 2009 and with great deal of suffering and death in 1918. One of the problems with our approach towards season seasonal influenza is that people don't take it very seriously, even though worldwide it kills about 500,000 deaths each year with three to five million cases of severe illness. The range of deaths in the United States is a mean much about 36,000 but it can go from as few as a few thousand to up to 49-50,000 the economic cost is extraordinary. The problem with the vaccine approach to influenza is we don't get excited about trying to bring the technology in to the 21st century until we get challenged by something that scares people. So you have a vaccine using antiquated technology of growing it in egg, is that has to be changed every year. That's not really a very good vaccine by the definition of what you need for vaccine. People keep thinking about the influenza pandemic of 118, 1919 then all of a sudden you get a pandemic. We prepared for the H5N1 luckily for us we put in to place a lot of approaches towards next generation vaccine for a virus that was going to come from the far east who the United States. In fact it went the other way. It came from the western himmers hemisphere and came as you know in 2009 with the H1N1. There were relatively few deaths deaths, it's very interesting, that's the reason why I love infectious diseases that pandemic comes bumps often the radar screen, the seasonal flu and seasonal flu killed more people than this pandemic flu. Only hitch in this was that it was particularly a bigger purred enburden in the pediatrics, it killed about three times more children in a regular year than with seasonal flu would do. The reason I show you these slides is because it was finally the wake-up call that maybe we should stop playing around with trying to get a new vaccine each year. So why don't we forget about seasonal flu versus pandemic just lump them all together as influenza that we don't have to worry from year to year which got a lot of people not to start to think, many of us have think thinking about this for years but this only became an important agenda item was to look for targets for universal influenza vaccine. There are a number of targets, some which would best be controlled by responses like matrix proteins and others are conserveed regions. That's what the VRC sets their cytes on and by the work of a number of laboratories identifying an epitope in the stem of the stem head component of heme gluten that actually was a highly -- in another very important paper from the VRC was of the induction of broadly knew for examplizing H1N1 influenza antibodies by using DNA prime and standard vaccine booster, vector boost that actually elicited antibodies that actually neutralized strains that you could go back as far as 1934 all the way up to 2007 that's really now one of the major goals of the non-HIV components of the vaccine research center. And in fact in this review that Gary and I wrote in "Nature medicine" just a few months ago we put together some of the thoughts that we'd been discuss discussing over a period of years which we both, all of us now at the VRC and elsewhere believe that this is something that actually is going to come about. I think the title that Gary chose, I give him credit because he chose that title is really very insightful. Because if the body can induce immune response in a natural way way, then what you really need to do is induce some unnatural immunity, maybe we're seeing a little bit that have with HIV also. I'm going to close with this last slide to again wish the VRC its leadership and its troops who are some of the finest in the world very happy 10th anniversary and I look forward to the continually learning from them, we have an interesting tradition, we every month or two or so I go over there sit down, they also have very good pizza over there. To sit down and learn exactly what they're doing and I can tell you every time I go over there I come out of that meeting very exhilarated because I always learn something new and get some important ideas. So thank you very much. Welcome to the symposium. [Applause] I am supposed to introduce the director of the VRC, Gary Nabel. [Applause] >> It's great to have you all here today and I thank everyone starting with Francis for his warm introduction and his support over the years and the support that we're going to need in the near future for our any budget. As well as Harold and Tony and so many people really. I think when I think back to what was happening more than a dozen years ago it really was kind of the combined wisdom of the biomedical research community, in addition to people like Bill Paul that Tony mentioned David Baltimore as the head of the AIDS research advisory committee, and just so many people that I don't have all the time to acknowledge but please understand that we're very grateful to everything that you do. Now, what I thought I would do in my talk is because Tony did such a wonderful job in giving you a swath of the history of the VRC to try to take a step back from where we have been with HIV/AIDS research, what we have learned in the last ten years and to give you that ten- ten-year perspective what we're dealing with with HIV/AIDS. First thing that I'd like to start with is that this is all about people. First of all it's foremost about the people that we're trying to help, those who are at risk for HIV infection and our ability to work in the laboratory to bring to the front lines to clinics, in this case, around the world the kinds of intervention, is that would make a difference. And secondly, when I talk about people in any research organization it really is the scientists in that research organization that make the difference. And I want to just say that this group has been together for ten years, I will refer to them individually as I move along, I do want to say that in many ways I feel scientists that we do stand on the shoulders of giants giants, that what we are able to do in immunology and virology and in translational research builds on so many years of basic research. And the only caveat I wanted to make about this, there is one person with is towering in this picture, Danny, and this is not because Danny is standing on the shoulders of John, he's standing on a waste can but really should not diminish from the tremendous contributions that Danny has made. I also want to use this to acknowledge not only the investigators but their spouses and significant others who have stood by us for these ten years and particularly my wife, Betsy, that Harold mentioned and it was in fact not easy to move from the cocoon that we had set up in Ann Arbor, Michigan, the same cocoon which Francis came. I know that was true for all of our investigators. So I just want to say that this is the group that's made things happen and deserves all the credit for the scientific advances that Tony has described to you that I'll describe in a little bit more. Taking the global perspective when we think about where we were in the early '60s, we didn't know that HIV/AIDS was in fact a disease and that in fact there were infections in different parts of the world. And in fact you'll hear lateer from BRTIS Hahn who has gone back and done molecular detective work of what was happening there. The virus was resident in the nonhuman primate population with crossovers at low frequency to human but at that point in time this was all the disease that there was on the planet. Now, what then happened is as we move forward you can see that the virus spread, we think originally spread out of Africa probably because of French speak speaking Haitians that were expelled from the Congo went back to Haiti, from Haiti was then transmitted to New York, San Francisco, also is then transmitted down in to southern Africa and in to Asia. What you can see is by the time we get to 2005 over 60 million people infected by the virus, more than 25 million deaths and if you again just look at the last decade, essentially the prevalence of the disease almost identical to where it was ten years ago. But you can see from the number of deaths, 45 million total versus 18 million that there are in fact on average acetone knee mentioned about 2. 64 million deaths per year. The incidence interestingly is very close to that. So one of the challenges that we have going forward is that essentially we are steady state with this disease. And if the number of deaths that we see is equal to the number of new cases then we've got to impact the number of new cases and that really points to the urgency for prevention. Either through vaccines or through some combination of preventive measures. Now, let me also take a moment to look back over the last ten years at some of the scientific advances and what we know about HIV/AIDS today that we didn't know ten years ago that will transform our ability to impact on the prevention priorities. So, I mentioned the work from Beatrice in fact very very fortunate that Beatrice will address us later in the session talking not only about the origins of HIV but also of malaria, obviously again one of the goals of the VRC to try to foster progress in some of the other major infectious disease killers, I think you'll find Beatrice's results from malaria as remarkable as those for HIV. The other major advance in understanding much the HIV biology in the last ten years has been the work that's come from the group, primarily George Shaw and Beatrice as well that described the fact that when people are infected with HIV there is largely a single founder virus. The founder transmitter virus is either clonal or oligo clonal . That's incredibly important insight in terms much understand understanding what we need to be preventing at the site of primary transmission of infection. And that site we now know much more about again thanks to the work of number of investigators here, Ashley Haase will talk in this meeting as well about the importance and mechanism of mucosal depletion, but several investigators in you'ding Mario and Danny and Rick and others have been informing us about out this occurs at virologic and immunologic level. You've heard a lot from Tony, I'm going to go in to it in another level of detail, how important the definition of atomic level structure for HIV envelope is. Essentially although we had a snapshot of what the viral envelope looks like ten years ago, it was literally that, a snapshot of one very specific form and through the work of a number of investigators, primarily Peter Kwong but Ian Wilson and others, Steve Harrison as well, we now know a lot more about what is happening at the atomic level with HIV. And more importantly or equally importantly because of that we now have an understanding of the structural and molecular basis of antibody neutralization. That is one of our holy grails of AIDS vaccine research. Then really the phenomenal find findings in the last couple of years from the VRC but also from Dennis Burton and Wayne and their colleagues where they have been able to identify exceptionally broadly neutralize neutralizing antibodies from HIV infected humans telling us that it's possible to generate the kinds of antibodies we need for protection in humans. For a vaccine that's an incredibly important scientific proof of concept. And finally, there really, we sometimes forget the importance of clinical research. And meaning the lessons we learn learned just from investigative trials of new interventions. And particularly in HIV there has been a number of very important studies on prevention modalities that include the trial that Tony mentioned as very seminal to the field as proof of concept. But also circumcision and pre preexposure prophylaxes. We'll have talk from Glenda gray later talking about some of the perspectives on doing clinical trials particularly in South Africa where the disease has taken such a toll. the role question, will these, how will these discoveries lead us to an AIDS vaccine. And what comes next. Let me then take you through for the next part of the talk just a brief sweep of how we're using structure get us to the endgame of the broadly neutralizeing antibody antigen . To put saw ziply. There are a large number of viruses, there are very view highly conserveed structures among those viruses and most natural immune responses to HIV will target a single virus but will not recognize the breadth of viruses that one will see that are naturally circulating in the world. So, how do we, given this genetic diversity as well as all of the other biochemical features of HIV envelope how do we focus in on the sites that are conserveed where we can generate those antibody. Here is video that was put together by Jonathan Stuckey in Peter's lab. What I'm showing here is a picture of the HIV envelope and this particular molecule in yellow is the CD4 molecule and what it's doing is it's inter- inter-acting with the CD4 binding site in HIV where it's making it's initial contact. And -- see if I can get this to keep going. I'm going to start it again. Here is the spike. Here comes in CD4 from your right. Making the contact, you will then see the impression of where CD4 makes that initial contact region on the trimer. Now extracted the trimer now have the monimer that is shown here and what I want to do in the next loop show you how some of the poorly neutralizeing antibodies inter-act with this critical region versus some of the very effective antibodies that can neutralizes lies the virus. Can see where B13 which is poor poorly neutralizeing antibody that when you look at that initial impression of where CD4 inter-acts with the HIV envelope that B13 is off target. And you can see that as -- these are the structures below. What you can see also is that F F105 which is also very poorly neutralizeing antibody that it is largely down and away from this major site of contact and some of what we've also learned is that in fact some of these antibodies like F105 can react with the monimer but not forms much the envelope that are in the trimer. So they're not reacting with the native protein structures. Here comes B12 the one that we did the initial structure that Tony mentioned and here you can see a fundamental difference which is the D12 is essentially hitting right within the confines much the CD4 binding site. It is relatively broad neutralizeing antibody. Not as good as the new ones what you can see is not only makes contact with this initial contact region but supersedes it it. And so here is the antibody inter-acting with the CD4 binding site you can see that it actually is doing it in a way that's very analogous to how CD4 does it. And in fact doing it more effectively at least at that initial contact region compared to CD4. That's why we think that antibodies like VRC01 can do what they do. Now, Tony mentioned the cores and stabilized structures that we have developed and we have used these now as tools to isolate these antibodies. That's what allowed us to identify was the restructureed score, is that allowed us to pick out these cells that inter- inter-acted. And when you look at the breadth of VCRC01 superimpose on the different families you can see that red indicates good neutralization, black almost no neutralization and green in between. This is from Wu from John's lab. You can see that VRC01 hitting more than 0% of strains and B12 which we used to think was great does pretty well against but not too well against the others. In terms much the -- what these antibodies mean for efforts in AIDS vaccine design they put us in a fundamentally different place from where we are in the reason -- essentially molecular level. The CD4 -- the VRC01 is inter- inter-acting with the beta 15 loop on HIV envelope almost identically to VR4 is mimicry that we any is important in achieving this level of recognition and breadth. Prior to 2009 in our ability to understand the biology of broadly neutralizeing antibodies I consider that to be the dark ages. We knew relatively little. We had come a long way from the late '9 Ross, we knew very little. When we were able to begin to identify antibodies like VRC01 we had new window and new opportunity to identify critical antibodies that had the kind of reactivity that we would want to see in -- elicited by an AIDS vaccine. Skip that. Now, one of the amazing technologies that we were able to take advantage of having identified the antibody, having the genetic sequence that we could go back in to the subject who gave rise to VRC 01 and do deep sequencing. Then we were able to do, this is largely the work of John and Peter and informatics group able to assemble these Dendrogr am, is that consisted of family members divide from the VRC01. Then we could ask whether VRC01 antibodies, we isolated another related to that in collaboration with others. so, that then told us that VRC01 was not a completely unique antibody. We could find antibodies similar to it in nature. Number two, we were able to identify antibodies from other donors and number three it also gave us much more robust way to now identify new broadly neutralizeing antibodies and monitor whether or not they would be produced after vaccination. So, what we have come to realize is that all of these antibodies do come from the 20 class they diversify. We can see range of antibodies, we have isolated now more than half dozen by the pro method. We think probably going to be dozens who perhaps hundreds more from the genomic method. What is remarkable, though, when we look at these we look at their ability to neutralize HIV that despite the fact they diversify in sequence they converge on this common shared epitope. It really is quite insight and really humbling in a way to sort of see how the immune system can use the power of genetics and biochemistry to basically create a solution to this problem in so many different ways. So the challenge for us now is how do we use this information to develop a vaccine. I should point out this is true not only for VRC01 but for the antibodies from the Scripps group, there are others coming out from other laboratories we now have whole set of tools where this kind of an approach can apply. So, the way I look at the problem now is that the VRC01 and first generation of exceptionally broadly neutralize neutralizing antibodies gave us a beautiful window on been we needed to go with antibody development. Having this new set of antibodies derived from genomic structures have changed the landscape. In fact in many ways I feel like we've been able to open up entirely new vista in identifying what those broadly neutralizeing antibodies are how we need to get there. Now, the question is, how do we get there? How do we get to the top of this peak? And one of the other insights from the diverse -- alternative antibodies that we have looked at has come from the analysis done by our bioinformatics group and it does look like there is a common evolutionary subtree for VRC01 from antibodies, even those that come from different individuals and that are quite distant in terms of their amino as it sequence. From this kind of analysis we can go back to the nodal point in those Dendrgrams what are the critical nodes, what is the critical tasks that we need to take from the germline to that set of thematically mature broadly neutralizeing antibodies . And Peter and Larry have put together now a set of what we think are nodal that go from the unmutated ancestor to mature forms. What some remarkable about this when you look structurally at where these changes on these antibodies seem to be evolving they largely are evolve knowledge in the areas that are making the critical context. Not a big surprise, but again insight that we didn't have and now we can use to our advantage because now we can begin to create immunogenerals that react collect sievely and morae efficiently with this particular particular -- and more efficiently with this particular path. Again to go back to the metaphor on the couple of slides ago, essentially if we're starting with the unmutated or germline ancestor, what we can now do is use the information from these inter immediate yachts to guide us to the point where we can generate the kind of antibody we want. I should emphasize, I think it's evident this is still in the hypothesis stage. But having a hypothesis for eliciting specific monoclonal antibodies is something new and something that we wouldn't have been able to do even a few years ago. So this information is vitally important to us, and meantime, we use this information and we use our knowledge of structures to design these immunogens and use these known structures to bind that those nodal intermediates. We've made some success in this regard. I'll show you one example where nonhuman primate we can use either specific trimers, I won't go in to the nature of these trimers here, me of this been extensively modifies or different prime boost combinations, in this case a trimer with an outer domain link linked to Chikungunya virus, that is not a vaccine. in the dotted lines are the negative control. In nonhuman primates we're able to generate these antibodies and they are very specific for that CD4 binding site epitope using the structure probe. We are making progress. These are not yet immunogen, is that can broadly neutralize but with the new tools the nodal intermediates and our ability to manipulate structures I think we're in the best position we've ever been in to solve that problem. Only point I want to make about this approach is that it's something that Tony touched on briefly and mentioned Stan and tremendous work that Stan has done over the years. Traditionally our ability to generate vaccine has really focused on our ability to identify sire row types. We had ha successful vaccine I think what we are dealing with a is new definition of serotypes. Not serotypes that we would be thinking about in terms of our natural immune response to the virus, but serotypes that are ee sec Shelly structural. That these are the highly conserveed structures that are there in the virus and we now need to create serotypes based on structure. And if we can do that, structures like the CD4 binding site, like some of the PG and 16 quantitynary structures, like the outer domain CG12-like structures that's really going to be our greatest opportunity in HIV for eliciting -- for generating successful vaccine. We're already at least at the beginning of that effort. Now, Tony mentioned universal fluid aside from everything else else. We found tremendous amount each year on seasonal vaccine production. Obviously with improved potency and breadth would increase public health and economic benefit. So we have been taking the lead from some very exciting papers that have been published by a number of laboratories, but among them has been this paper from many, as well as Ian Wilson who solved this structure and basically this is the antibody structure of a broadly neutralizeing anti-flu antibody directed at the stem region of influenza. So here is the stalk, the stem, here is the heme gluten and Ian will talk about this he's really been the leader influences structural biology in understanding of neutralization, been at it sincement late '8 Ross all hear about it from the master. We've been able to take sad advantage of -- advantage of this knowledge to design probes where we can then use different vaccine platforms to try to elicit these responses. And again what we had been able to do, Tony mentioned our earlier work where we found that if you use a prime boost combination of DNA prime mate with vaccine boosting we could improve the heme glut ten inhibition response. We've taken this in to humans, interestingly this shows you the importance much doing clinical trials. What we learn asked that if we have only a one month interval between primeing and boosting there's no increase in the HAI tighter F. we're patient we wait six months you can see that DNA primeing actually significantly enhances the boost or significantly enhances the antibody response that would -- profession against in this case virus. That anti-stem antibodies block HAI as others don't. So, at least now we know that for flu as proof of concept we can generate antibodies to the stem region that should have the kind of breadth that we hope. I think as to the general optimism that we had this may be a feasible approach in the future. Tony mentioned chikungunya we have developed a vaccine that is now moving in to phase one trials. We have a -- with our collaborators at Merck I believe Danny who is the P.PI from Merck is here with us today we're trying to develop Chik particle that can be used in southeast Asia we're moving forward on those. The good news is that we've been able to adapt this technology to other alpha viruses, eastern and western eke wine encephalitis, Venezuelan, we think it's quite possible that we might be able to generate a broad pan alpha virus vaccine with this platform or with the next generation of it that we hope to put together with our colleagues at Merck. In addition to all that we've been using the chikungunya parcels as scaffold to present HIV epitopes we can actually insert them in to the spikes of chikungunya virus, we can actually create insertions here we've done with this Michael, you can see some of the chikungunya particles where HIV envelopes has been inserted here in dark blue. You can do different subscription you can see the ODsDs that are now arrayed on to the surface of this virus-like particle. Presents highly my tricks -- matrix antigen this is another way by which we can stimulate strong immune responses. Again the complementary of the different vaccine approaches is very important. I think we learn from one how it can benefit the other. I won't say anything about E bola vaccines other than we now are back on track and have some candidates that we'll be moving forward in to the clinic. Finally I just want to end with another reminder of how important it is for VRC I think in general in the entire realm of biomedicine research to be able to translate from the laboratory in to the clinic, Barney Graham runs our clinical trials unit and this actually shows the number of subjects that Barney has enrolled over the last ten years of the VRC up to near 5,000 subjects. And whole range of different diseases. I also want to mention that among them we are now working with Scott Hammer the principle investigator and Larry who is the HVTN principle investigator to test DNA prime add boost, prime boost combination in the HVTN505 trial that is in the midst of enrollment. Don't have anything to say. But the only way you can ever tell if these vaccines are going to work is to do the efficacy studies and these are tight efficacy studies to give us endpoints as efficiently as possible. In addition to the vaccine studies now that we have very good anti-HIV neutralizing antibodies, we've begun to look at other prevention strategies with antibodies against HIV, passive prevention, you'll hear fascinating talk from David Baltimore on the future of antibody prevention for HIV and some of the remarkable tools we now have to pursue this in very creative ways to use it for therapy and maybe even to go after eradication of the reservoir. This slide really just reminds me to say that we are pursuing initial studies in humans using the antibody flow path of transfer then like to see whether it might be -- if it works whether it my pave the way for Gene-based delivery of antibodies that can can used in the knew tour. So this is the model that we use at the VRC. we translate in to the clinic. Generate improved product. We're very, very lucky to have the support of NIH, not only here on campus but at some of our contract facilities in the clinical center. We have an immune assessment laboratory as I mentioned it's really the people who make this work. And I'm so grateful to all of you who have helped make this a working center. I'm hopeful that our goals and aspirations for highly successful AIDS vaccine and other preventive measures through other infectious diseases will become a reality soon. Thank you. [Applause] . >> I'm going to hand podium over to John Mascola by trusted and valued deputy who will moderate the rest of this session. . >> We will hear from some of our distinguished guest speakers. So it's my pleasure to introduce our first peeker, he is Dr. Ian Wilson from the department of molecular my polling at the Scripps research. One of the most distinguished and -- focused on understanding the structure and function of the molecule. His laboratory solved probably too numerous to count, crystal structures, T-cell molecules, ligand receptors and many antibody molecules. He has log been major contribute tore our understanding of HIV antibody structures and function but today actually going to talk about influenza antibodies and implications of that work for influenza vaccine. >> Thank you very much, John. Thank you Vern much for inviting me and having the privilege of talking at this symposium. I visit the VRC quite frequently, many times a year. It's been great seeing the development and progress and particularly inter-acting with Peter and with John over the years. Let me talk today about flu virus and implications that we offer universal vaccine. Happy of the things that we've heard about sieve have been influenza field with the discovery of neutralizing antibodies to flu. Really quite surprising that up until only about four years ago we really didn't have any neutralizing antibodies for flu so this is opened up tremendous new opportunities now for thinking abut how we might approach this universal vaccine. So, let me say couple things about influenza virus that to get us started, it's a negative stranded virus, eight strand of RNA, two surface proteins, and I'm going to focus today on influenza A because that's one that causes major pan them I can and epidemics. This is prevalent in humans, birds and pigs and these viruses, reservoir for most of the flu viruses is in aquatic birds, 16 types in birds at hemagglutinin and nine others. Only three have appeared in human population and remain resident for H1R and H3. It's going to be very important for discussion I'm going to have that these types can be divided in to two groups. Group one and two. Ten of the say types are in two. So, we know that this is the one of the reservoirs that we have is in birds, we know recently that there's other possibilities of what can happen when you encounter pigs. So, normally when we get a human virus, basically when we get new pandemic it's a mixing of these different genes through human virus and bird virus with pigs as potential reservoir. Outcomes these completely new viruses, mainly have different surface antigens almost always the hemagglutinin hat changed and one of the polymerase genes. Constant evolving code. Once you got that pandemic in the population then seems to have unlimited ability to mutate. We've seen H3 since 1968 now and still going strong. This is what has happened over the last hundred years, clearly we had pandemics before that. We had H1N1 introduced in 1918, transplanted by H2N in 1957. By H3 in 1968. Then we had reintroduction of epidemic in pan them I can of H1N1 in 1977, then we know big scare about bird flu H5N1 in the late '90s, early 2000s then twine flu rereintroduction very odd having circulating at the same time in addition to H3N2. So as been mentioned previously nice to not have new vaccine every year. If we can have one even last for one subtype let alone all of these different subtypes. This is the question that we are trying to focus on is can we get a broad neutralizing antibodies response generated against all subtypes or at least many. Prevent virus getting in receptor for influenza virus which is the heme flute anyone. Most of the antibodies are to the head region that's where the receptor is bound. That's what most of the antibodies do block this virus getting in. More recently started to see antibodies to the stem region that suggested that we can stop this fusion activity and so these antibodies act by prevent preventing, uncoil of the virus and ability of the virus to put genetic material inside the cell. But by circulating around receptor binding site by blocking it in that way. Fusion antibodies, once that block fusion are rare. Activation even error, for flu not many other sites that are documenteddocumented. This is the situation that we've been in with flu that most of the antibodies that we have are strain specific they're quite abundant which means we have to get a new vaccine every year. We haven't even had until very recently ones which are subtype specific, that is the -- let alone ones which can bind across subtype. We're starting to see some indication that the maybe some hope that we're getting at some antibodies now that can cross these two major groups that I showed. Good nice is that we had really none of these types of antibodies as quite surprising, we've been following HIV field as of late in this way. Flu has been a bit further behind. We haven't really had antibodies in these 2, 3, 4 categories since the last few years. But seems now that flood gates are opened and we're getting much of more of these type of antibodies being produced. So the first indication that we could get neutralizing antibodies came from these three sets of papers, one from our own institute with Richard in collaboration. One from collaborators. One from Harvard. All of these antibodies could -- it wasn't obvious that they were the same. But it turned out that they actually were. They were all one particular sub family VH169. All isolated using agents and recombinant HAs to select. These were the sequences that were found between these different antibodies. As you can see they're very similar to one another, in fact remarkably close to the germline. Very few, this is again very unusual compared to say HIV antibodies. They're very few mutations from germline. These are the Crucell down here, Marasco down here. To say that the very few differences. What's important about these -- this particular germline family are these high trophy I can residues. At this stage we got in touch with Crucell and started collaboration, this is their data from their paper. Showing basically these antibodies was very exciting time because these antibodies for the first time could be shown to actually neutralize a large number of members of the family of this group one didn't touch group two. What is also important is this neutralization was very strong against different subtypes. You can see that we have affinity against multiple different subtypes. Crucelllo showed protect mice and also importantly was difficult to generate the escape mute tants. We got the antibodies from -- the started working on he produced the hemagglutinin and got the crystals got these two crystal structures this was with H1N1, this is with H5N1 so these are the two different subtypes. You can see the antibody here in orange and yellow and for first time the binding in the stem regions all the way down here. Didn't bind up in the head region where most of the other antibodies strain specific ones had been. This was work couple much years ago to update you on that, we've solved a number of different other antibody structures, this was the original one, this is one from others, this is not our structure, this is one from Wayne's lab. Remarkably all of these antibodies behind in exactly the same location on the surface. So look at the -- overlap of that, you can see these three antibodies, the two Crucell bind in exactly the same orientation, about 20 degree variation. But important component is that this H appear, base of H3 are really same meanting things. There's a little walking around, flexibility that's allowed for these antibodies recognizing very much at the same epitope. Why VH169? Well, that is hydrophobic groups on the end of the loop here, in this case it's -- variable regions find this hydrophobic pocket on this stem around diffusion binding -- of the fusion pocket. So, what we can see is that this insertion to that really helps select at that particular family of antibodies. How did it work? Well, I'm not going to show you the primary data but a lot of data to show that first of all the structures that we determined were below Ph 5.5 which is usually Ph of activation going to the fusion active form. So normally for virus to get in this is the neutral Ph form, when you go to low pH you get series of confirmational changes that lead to fusion. Of the viral membrane and whole cell membrane. It 'fierce that -- did a crystal structure say Ph we got this structure this suggested that the antibodies were but trussing in preventing this. The hemagglutinin becomes very sensitive to trips Lynn. When you put the antibody on to that -- there was no trip is in tense sift. What the trypsin -- design mol calm to the same epitope. He took the epitope he base you cannily started putting site chains what you thought were energetic hot spots. He then tried to figure out let's go through try to figure out from all the 50,000 or so structures in the PDB how you might attach these. In doing so he generated some basically new proteins that could bind to this particular epitope. He did about, almost 100 of these. He started off with two designs that he tried to improve, one initial KD of 200 nanomolar. He ended up immunogenesis of these and three nanomolar for two of his designs. These as you can see are on different frame works, these site chains, hot spot site chains were the key components for these particular templates. This was work done by David's lab. In my lab, crystal structures of this antibody -- sorry of this small molecule, small protein molecule. Predicted how it would bind, predicted structure shown in blue here, crystal structure shown in red and you can see it's remarkable how well they designed this particular molecule. Even to the level that side chains even got pretty much almost perfect. This was pretty exciting. This protein molecule bound, had similar characteristics of group one and not two. Also prevented this Ph trypsin digest at low Ph compared to not having it. And so these design properties really for this small molecule were very similar to the original 6261 antibody have some potential for development. I didn't address the question yet, why did these only bind to group one? One of the main problems with combining to group two because epitope was largely conserved in group two. Is that there's glycosylation site in the way. A question that sort of then became an issue, particularly for Crucell was to find out is there another conserveed stem site for group two. The answer is, yes. Crucelell ice laid group 23 stem antibodies, these are two of the antibodies 8020 and 804 that neutralize multiple distinct strains. and these antibodies do something else which is very interesting, they block HA1 and HA as well as Ph dependent change. So again in collaboration with Crucell we actually, Damion did the structure and this is 020. And this is where 6261 you can see that pretty much they're nonoverlapping at this binding much further down, this would be the membrane down here, this is facing away from the membrane. Then color coded here are the epitopes. You can see that there's only really two in common between 6261 epitope and the 020. So really defining completely different sites. Coming in this direction where as this was come in over here. The antibodies as I said team to be able to block the pH fusion activation. Binding very close to, this is the fusion peptide coming out here this is only subset of the bottom part of the stem. And you can see that bind on to the end of this loop from the fusion peptide loop. Also bind to the edge of this sheet, one of the inter-actions is shown here with the CDRH3 inter-acting with the sequence independent way to the main chain here of this strand. Why might also block this cleavage activation from HA0 to HA1 and HA you can see the blocking again very less to where this residue here, last residue of HA1 used to be attached to HA2. These antibodies also defer from one type specific 1s which were largely heavy chain binders. These use all of the CDRs and you can see that they're scattered over the surface of the stem here. This epitope is highly conserved also if you go to the fusion active form these residues which are down here are very important thought to be infusion mechanism for capping the end of these here, this long helix, again it explains sort of the activity that we can see for those antibodies. Same question can be asked now, why don't those antibodies bind to group one. Again epitope isn't quite as conserved as we saw from the epitope in 6 61 for crossing these different groups. But there's -- it is largely conserved main problem again that there's carbohydrate in the way in group 1 type hemagglutinin compared to group two. This blocks the site. So this suggests then that you can get some sort of universal mob clonal antibody based cocktail. I got these slides from Chris. We know that we can antibodies now combine to these human viruses, the group one and group two so largely viruses that we're interested in can be bound by these two different types of antibodies. We're now down to two different antibodies that can actually neutralize all of the important viruses. Let me finish off then with the last antibodies class that I'm going to show you. This is new antibodies that we got recently from Richard Learner this does neutralize across groups one and two. Combine those that we've tested so far. So this is another very interesting virus, the question is, where does this bind? Is it going to bind in the same region or differently. Damion has done the structure of this, this is our collaborators the Scripps, so Damion has done the structure of this in two different forms alls HA1 and trimer. The antibodies bind up in the head region here. Only reason is receptor binding site. So we'll see from this movie here, here is the antibody coming in, it's binding right in there, it's actually put in this really long CDR43 right in to the receptor binding site. This is the first antibody that we've seen, despite B12 that have been around -- B12 for awhile. Has been fairly common for HIV to be able to get in to the receptor binding site. Much more difficult for flu, much more shallow and so it's difficult to actually get enough surface to grab on to to these rather limited number of conserved residues in here. The way it's managed to do that by inserting this long CDRH3 in Lear then having extension on the end some inserted residue in the end of H1 to be able to reach down and touch the top of that helix. So this is more cartoon version of it. We've got five residue insertion in here, got 25 residue long CDRH3 that can reach down get in to this particular site. This is last I'll show you of this. Showing this long CDRH 3com and snaking around coming back out through receptor binding site. This is the view of the binding site for hemagglutinin, it binds carbohydrates, connected to glak toes then glucosamine and so you can see that this is a pretty small surface area this managed to sort of come in and out there, this loop comes down sits down on the top here. I'll finish off there just say that this is pretty exciting times for flu. Because we've now got neutralizing antibodies that can now span some types, any mention those. Number much other antibodies that can go and basically neutralize almost everything within particular subtype. I've shown you antibodies to group one, group two now showing you ones which have limited activity but across groups. These are combined to the stem, this winds up here to the receptor binding site. This is the final slide, a lot of the work that I've shown here was done by, as I said this very talented grad Walt student. Funding from NIH, from Crucell and NIAID and consortium with NIAID. So this is also collaboration, acknowledgement from Crucell for the work that I've shown. [APPLAUSE] >> That was fantastic, thank you very much. We have one more of our guest speakers before the break. Then we'll pick it up after that. My pleasure next to introduce boatrace Hahn professor of medicine and microbiology at the university of Alabama Birmingham. She spent time here at NIH in early part of her career well-known to many of us for her many contributions for related to mow creek could you already biology of HIV including first isolation of molecular clones of HIV including tracing the origin of HIV 1 and HIV in African chimpanzees and more recently understanding the pathogenesis of SIV in wild species of chimpanzees. She's recently turned her attention as Gary mentioned to molecular genetics. >> Thank you very much. It's great to be here. When I saw the program couple weeks ago, Gary, my talk sticks out like a sore thumb. It has absolutely nothing to do with vaccine development. He said, no, no, don't wore we, people want to know where the stuff is coming from. But he really dying to hear the malaria score rebecause he's thinking once HIV is done maybe we move ton to new field. I'm all with you. So, with vaccine known that AIDS is -- we've known it since basically sequence of these viruses were determined. Just couple years after HIV was first discovered the reason being that there were two AIDS viruses causing similar diseases in humans but being quite different genetically following within cluster of viruses that we now know are primate lent viruses. Lentiviruses which appears in macaques and led way to other viruses came from the original of HIV 2 became clear because they were in same area where HIV 2 was endemic. The origin of HIV 1 was more difficult to determine for two reasons. There was virus closely related to HIV 1 which was identified in handful of chimpanzees. But there were two problems, one when people looked in captive chimpanzees for other viruses they couldn't find them. Over 2,000 in U.S. primate centers and Europe there was only one that could be found. Second early viruses there were viruses were sort of virgin they could have been similar to any one of these HIVs the question was, whether it was another primate species that was responsible. What it really took was develop noninvasive detection characterization. You have people inspired us to do this, we went to the place for many -- one of the students presented work how she isolated viral sequences from the stool samples of babies. I remember thinking to myself, why in the world would you ever do that. Anyway, 20 years later that's what I'm doing. End up having fun doing it. [laughter] We collect fecal samples throughout Africa, in fact our collaborators, these are quite useful because you can use them diagnostically, strained follow viral specific antibodies. Also extract nucleic acids and do analysis of the host DNA or viral RNA in the case of SIV. We found that the fecal samples are useful to study other viral pathogens and parasites. Spebd this is summary of the last ten years of work. Molecular immunology looking at 7,000 fecal samples collected at 90 field sites throughout central Africa this is what we see. The sites in yellow mean that Siv is there and us you can see S.I.V. is endemic in central Africa. However, the virus absent from two other subspecies of the common chimp, here Nigerian chimp and here, the western chimp. That is explained why the search initially for S.I.V. was unsuccessful. The subspecies doesn't have the virus. Interestingly SIV CP Z is absent from the second species of chimpanzees. The distribution as you can see clearly there's distribution is uneven with areas very high prevalence in seethe earn Cameroon where prevalence can reach up to 30 in coninstance 50% but other areas of prevalence lower and some areas have no infection. Does this make sense? Yes, it does make sense when we consider the origin of SIV CP Z which is recombinant of viruses that in -- chimpanzees 'required by cross species transmission. It's important to remember that all SIV strains have same identical mosaic genome structure. Again, it explains why we see it in only two of the two subtypes. We think that the cross species transmission occurred relatively more recently. And natural ranges of the monkeys that contributed to SIVs are here in Cameroon and surrounding countries. We think cross species transmission occurred some time here then virus spread to eastern chimps before or after subspecies had separated. However it never spread to the other two subspecies. Now, the uneven distribution we may also have explanation for that. Recently presented at the meeting turns out that the chimpanzee CD4 molecule highly polymorphic unlike that of gorillas there is second site in the region that in -- it's very hard to infect in vitro we've never found naturally infected chimp with that genotype. In fact over 100 chimp, is that we now know are infected we have done genetics on their fecal samples only three have this glue Sol sakes site. Then they have additional changes that must compensatory. 3,fecal samples from gorillas later at 30 field sites we know distribution of this virus. SIV is endemic in western gorillas but absent from eastern gorillas these are two separate species of gorillas. We are also know from genetic analysis that sieve resulted from transition of chimpanzees, there was transmission here from chimps to western gorillas. The MR CA calculation suggests that the introduction occurred about 100 to 200 years ago this didn't happen yesterday, however sievegor much less -- SIVgor is much less common. About 1% a at these sites raging from zero to 5% although there have been identified hot spite here in southwestern Cameroon where prevalence have gone up to 25%. Hard to say whether SIVgor is on it's way in or on its way out. Now when all these sequences are put together in a tree that also includes the human and gorilla viruses, couple things become apparent. First of all, there are two very distinct lineages, those viruses here in black infecting the central chimp, to go diets. Those are here infecting these. When you see this of course you know there's not only one HIV strain but actually four, group M, the pandemic version, group N, O and P were discovered in succession. All of these sequences were in the radiation of the central chimps. Indicating that it was the central chimp, is that were the original reservoir for these infections including the door rim la viruses. Now, we know that for example can the M group has very, very close relatives in chimp viruses in southeastern Cameroon so can pinpoint the origin of group M to that geographic region same true for group M very close relatives from south central Cameroon in the forest area. We do not have such close relatives for groups O and P. And for gorilla viruses, even for group O we still don't know whether the origin was chimp or gorilla it depends when the chimp transmitted their virus to gorilla if it happened on this branch then the group O is a gorilla virus if it happened in this branch then group O is chimpanzee virus. We know that group P is of gorilla origin. Both chimps and gorillas have transmitted to humans. However the spread of the cross species transmission has been variable. You've heard group M has gone pandemic now almost 70 million people infected with this particular lineage of HIV 1. The other introductions have remained relatively rare. Group O has some epidemic spread primarily in Cameroon, Nigeria and neighboring countries. Other introduction group N less than 20 people known to be infected. With group N there are only two individuals now identified to have group P. Again SIVgor is relatively rare. Most surprisingly there is not a single human or eastern door rim la case of transmission of virus. It may be that these are actually two divergent to be able to adapt to these hosts. I'll get in to that a little later. Now there are a number of questions people frequently ask. One is, how did it happen. From the biology of the virus as we know transmission must have occurred thrummy cues us membrane to infected blood or body fluids that happens most frequently in the context of bush meat hunting. That is so far hypothesis. Where did it happen? We know where closest relative of the M group infect chimpanzees in southeast Cameroon. Where the closest relative in south central Cameroon. We don't know that wet for groups O and P. When did it happen? I think we have very good data now saying that HIV 1 group M pandemic emerged around 1920. There are viruses if you will from 1960s that have helped calibrating this clock similarly for group O we think emerged around the same time. For group M&P it's difficult because there are fewer viruses to look at. It appears that they emerged later. The last common ancestor for group M is calculated back in the 1960s but that remains with a question mark because it -- fewer viruses we have less reliable are these calculations. Where did group M start to really get going? I think it's clear now that early diversification of group M took place not in southeastern Cameroon where the chimpanzee reservoir is but in and around, at that time recalled Leapoldville where they have did discovered. Even more exported to other parts of the world. Why was AIDS not described until 1981? Because of the expo ten shall growth. Basically the virus was in people for 60 to 80 years without being recognized only around time, mid 1980s, 1990s were there any clinical cases to come to the forefront and remarkable that the virus, is that were identified by others in the 1960s we would estimate at that time there were only about 4,000 cases they were quite lucky to have found these viruses. Why did AIDS not emerge until the 20th century. That is where speculation begins . Many have implicated the rapid growth of cities at that time in that part of Africa. With it came destabilization of social structures there. Is a paper that shows that there was greatly increased prevalence of STDs at the time that may have helped transmission. French group that has investigated large scale injection campaign at that time. All of these things could have helped to jumpstart the AIDS epidemic, whether it did or not I don't think we'll ever know. I've shown you where the human viruss came from I want to point out there are 40 other primate species that are naturally infected with I.V. in those that have ha been studied primary three the African monkeys the virus teams nobody not pathogenic for the host. The great apes, both chimpanzee a and gorilla is where it's pathogenic in these species. There is just no way to study naturally infected gorillas in the wild. We were fortunate to be able to study natural plea infected wild living chimpanzees we started to do this in 2000 in the national park with the support of Jane Goodall, they are completely wild but have been studied since the 1960s and their life history, social structure, behavior is all well-known. We superimposed our noninvasive viral diagnostics when we put all this together we realized that HIV infected chimps have 10 to 16 fold risk of death that's quite high. HIV 1 depending on the cohort would be anywhere from 15-60 fold. It seems to be sort of in the middle there. Infected females ever less likely to give birth. Infected females have much higher infant mortality. This is also true of HIV 1 infected women. And another group we showed that infected chimpanzees actually can develop CD4T cell deplex and histopathological findings one SYS ant with end stage AIDS. We have two such cases. And even discovered virus in the brain of one of the chimps who died with such a syndrome. We also documented one community in Kalande community with a high prevalence of 40-50% suffered recent catastrophic population decline going from around 45 individuals to less than 10. So clearly AIDS has afflicted chimpanzees long before it did humans. One word about HIV, I said it is -- HIV 2 originated in west Africa, importantly to note again the virus weren't only transmitted once but in this case on eight different occasions only one of these transmissions resulted in epidemically spreading pathogen which is referred to as HIV group A. The others were either -- had very limited secondary spread or no was viruses identified in individuals. So, we now have 12 SIV transfers from two of the 40 primate species. To humans. Only one of which really caused an AIDS pandemic. How can we explain that? Well I think innate host restriction factors are probably largely responsible for that and there are number of restrictions that have been described and others that are being discovered as we speak. But probably has greatest impact at least on the spread of HIV 1 and HIV. Protein that is in the Miami viruses have evolved to counteract tetherin. Lentiviruses use VPU. Here in the membrane or the inter-act on the cytoplasmic tail. They bind to tetherin and purple it off the surface cannot do its job. It is instructive to look what happens as viruses went from monkeys to AIDS to humans. Focus on the chimp. I told you chimpanzee viruses mosaic, and resulted from cross species transmission of viruses from monkeys viruses which brought VPU in and they brought -- when cross species the VPU in officially probably didn't work very well in ham pansies. Adaptive process was required and at the end NEF was the main counter active of tetherin in the chimpanzee. When virus was transmitted to Gore hails again initially probably -- Gore hail, because of the the piece -- either VPU or NEF were active but eventually adaptation selected as counteracting viral protein. Now, when the virus then went to humans it was different story because as it turns out human tetherin has deletion right here at the spot where the SIV inter-act with this protein. When the chimpanzee virus went in to humans neither VPU or -- because of being one that contracts chimpanzee, VPU lost the function come from the greater monkey virus. What needed to be done was these viruses had to reacquire tetherin activity and in the case of group M it was VPU that had to gain this function, regain this function. As it turns out this didn't work very well with group P and O and it only worked partially with the M group. Now when the Magabeys generated, the virus uses Nef it doesn't work because of the deletion for HIV group A came in. The protein that counteracted tetherin. No one has looked at the other transmission. Really when you look at all of this, the only one who really succeeded to counteract tetherin is HIV 1 group M. That is only one that has gone pandemic. So in conclusion, AIDS caused by two lentiviruses, HIV 1 has resulted from four and HIV 4 from cross transmission. Outcomes of these have been highly variable. Ranging from really sporadic infections with no secondary spread to fan Del I can you need inflammation there you need activation there, to overcome these host restriction factors to replicate even under the circumstances introducing mutation you need in order to adapt. What does this happen to do with HIV vac seep development. Perhaps I showed you what tetherin does and it is possible that lack of effective anti-tetherin activity may result in reduced virus squeaks. May explain why the sporadic groups of HIV 1 and were able to first infect humans to make the cross species jump but then were unable to establish an epidemically infection if this is true perhaps focus on the VTU tetherin inter-action somehow put this in to our vaccine design strategy. Teeth run seems to be a big hurled that the virus needs to overcome. Not just at the point of cross species transmission but perhaps also later. Now, going from the origin of AIDS to the origin of malaria, you may say, what in the world is she doing with malaria? The reason is simple. We did our study in Gombe found out that it is quite pathogenic causing AIDS. And because that have we always had an ear out for comorbid infections, we were watching these papers coming out describing new parasites in great apes in captivity and then also in the wild but at that point there was nothing we could do bought all the studies were done with blood samples. French group published a paper collaborating with Martin Peters and her husband showed that in fact you could diagnose Plasmodui America infection with fecal samples. We knew we were in business. Just to recap a little bit. Of course malaria is a parasite, it has a very complex life cycle, there's a mosquitoe that transmits it, a blood stage there's a liver stage and sexual reproduction of the parasite happens in the mosquitoe. There are five malaria parasites that infect humans, probably worst is Falciparum. Vivax is in Asia. And other parts of the world as I will show you. It is probably the most fascinating malaria parasite. In addition there is plasmodium is in southeast Asia in macaques several thousand humans become infected and macaque parasite and inflicts quite a bit of morbidity. Of course there's no vaccine and widespread drug resistance. So, origin of Falciparu, some of interest. Only one relative to human people that is strain. That was I'd tied in captive him chimp. Then these two strains were put in to a subgroup which was called Laverania there were quite distant from the rest of the human and other primate and rodent and bird parasites which are shown here. Here is Vivax and here is malaria and others. There are number of other primates and rodents and birds that are infected with these parasites. Because this tree somewhat looks reminiscent much the tree of the hosts themselves, many people have believed for long time that there was co-evolution of parasite in plasmodium parasites to be over a hundred million years old. Looking, probably the last common ancestors of human and chimps was infected with the last common ancestor of their parasite. So they thought there is too little diversity to explain six million years, they propose a much more recent origin, malaria was published and he thought it's more likely about 5,000 to 10,000 years ago because that coincides with the onset of agriculture when human gatherers and hunters were coming together and living in villages in a greater density, it's also time when mosquitoes species started to diversify. As additional I months mode yum were discovered people proposed various origins for P. falciparum. But there were problems with these studies. First of all, almost all of the Sam peps were analyzed from captive apes in all instances except for this study from blood samples. Very limited sample size. Also as it turns out apes like humans are infected with multiple species you generate artifacts in vitro. You generate recombinants. The solutions were to use neckal samples from wild apes and look across central Africa at multiple field sites. The wrecks looks approach that ensures that at the end of the day you amplify a single genome. So you avoid PCR artifacts that are coming from the fact that the polymerase jumps between templates you also avoid PCR substitutions. We used this approach initially just targeting a small -- in the plasmodium mitochondrial DNA which is about six KB long. Using single genome application we also targeted applicable Gene, another Oregonal that the has and LDH. Initially we screened, went to our freezers pulled out the same samples we jeweled for SIV diagnostics we found that ape plasmodium infections were widespread in all four subspecies, we found it in western gorillas and here are the prevalence rates indicated that we estimated based on fecal detection method. They were pretty high, most likely a underestimate anyway because very likely that the detection sensitivity in fecal samples is below that of blood. I would venture to guess that most of these populations are highly endemically infected at very high prevalence perhaps reaching 100%. Interestingly eastern Gore hails we did not find any infections. This is what we found. We found six lineages which basically represents species, three of them infect chimpanzees. Three of them exclusively infect gorillas, G1, G2, G3, human P. falciparum fell right in. All of the human strains in the database at the time did. This is just a blow up of the G1, here you see the human parasites falling within the radiation of the gorilla parasites forming single lineage which suggests single introduction from gorilla is human. Also supporting the direction of transmission from gorilla to human is diversity in the -- which is much greater in the gorilla parasites than it is in the human parasites which is what you would expect if the transmission direction was from Gore hail is human. We went on. The G1 clade with the gorilla parasite either looking at the coding region, these are protein sequences or nonoverlapping nucleotied sequences on the other side of the might condecreial. -- within the gorilla clade these are all the human haplotypes in the database at the time. There were 105 and here the same for nucleotype sequences indicating cross species transmissions from gorillas to humans. Interestingly one of the previous stud he's had implicated the origin because found viruses very closely related to human P. falciparum. You see where it's clustered right in the human clade. These parasites also had drug resistance mutations that were prevalent in the local human population, what happened here although they are not infected in the wild they must have acquired human falciparum in an Tift. Some argued that 105 full length mitochondrial are not sufficient to make the claim that the door rim la was the original reservoir. In particular since very few of these sequences came from Africa. We have since increased sample size where sequenced almost 600 additional samples we got them from our collaborator at the Sanger institute. 35354. Through Martin Peters, 220-some samples from Cameroon. They follow just the same as before, both in the radiation of the gorilla parasite there's no doubt that the direction of transmission went from gorilla to human not the other way around. This is summary now of all the different genomic regions. There are six claves or species which we have given some names to then there were also some non--- which I'll get to. Conclusion at this point that chimps and western gorilla each naturally infected with three Laveragian inspected. Almost comprised of mixed parasite infections. Humans acquired P. falciparum following an event. The time of cross pe sees transmission is unknown. There are very calculation but all of them have various assumption either assumption of virus host coevolution or assumption of when cross species transmission might have occurred we have no way to calibrate the clock. Like we do for HIV. So the cross species transmission could have occurred anywhere between 3,000 and 3030,000 years ago. Falciparum have been found in captive Bonobos and chimps of very recent human origin. As it turns out apes do not only have falciparum like parasites but parasites that are genetically closely related to human situations. May be more species out there in wild apes that have been found in humans. One of these is most interesting that is plasmodium Vivax. It's endemic in Asia and South America. Until recently the closest relative to plasmodium Vivax has been found in macaques in southeast Asia. These parasites right here putting those two things together, investigators have promossed that the origin of P. vivax is in Asian macaque. However -- this is the distribution of fall falciparum versus vivax when you compare the two, they're pretty well superimposable except for west and west central Africa, P. vivax is absent from that region. There is a rope for that, because -- that reason coincides with the distribution of red blood cell negativity in the human population. Turns out the chemo tine receptor is the tells that P. vivax infects there. Is mutation in the promoteer that prevents the expression ever the Duffy antigen, vivax cannot infect humans and that's the reason why in this region of Africa there is no P. vivax. People wondered why this genetic change was selected to completion. Because obvious Levi vac is absent, currently not present in central or west Africa. In addition the tropical reference laboratory in London noted about 1% of travelers returning from central Africa have P. vie racks yet there is -- vivax yet there is no P. vivax in human in that region, there was recent study that said 400 humans in the republic of Congo were reported to have antibodies to P. vivax. We were wondering where is the reservoir for all of these infections. So, we pulled our fecal samples out once again, and looked for the distribution of P. vivax in African apes. It is certainly there. We tested over 7,000 samples from the same field sites using diagnostic PCR. I should add that finding P. vivax in fecal samples the two orders of magnitude more difficult F. we look at 100 fecal cam sells -- samples find 20 to 30 to be positive for Lavariarina with the fact that vivax infects reparticular could you already sites there for is present in blood sample much lower parasites loads. Nonetheless, we did document vivax infection in all chimpanzee subspecies that we looked at. We also found it in western and eastern gorillas, however we still haven't found it in Bonobos. Overall, P. vivax is less common than Levarania, we found it in eastern gorillas and also found it in southeastern Tanzania where we had not found it before. Putting sequences that we found in to the tree gives us this picture. This is very preliminary. Here in black are all human P. vivax haplotypes that are available in the database, almost positive 0. Almost 300. In green are the gorilla parasites. There is gorilla and chimpanzee parasites out flank human parasites, very similar to what I showed you before. But preliminary because this is based only on 3BK mitochondrial there isn't the specific difference as we saw for Lereraniga they are in per spaced. Very closely related to the human parasites. The question rises whether this is all one species and question also arises, what is the direction of transmission, we cannot be sure yet. These two parasites were reported by another group. As turns out this one was identified in wild chimpanzee in the national forest, the group with our ape P. vivax. This one was identified in sanctuary in the DRC it grouped with our human parasite. Also interestingly, as we were doing this we found a new clade which possibly also in African apes and two chimp which may represent a new species. In conclusion vivax-like parasites affect -- vivax is difficult to detect in fecal samples most Lukely because of lower systemic parasites. Commonly in mixed species very represent the same see. Origin of human P. vivax still remains to be determined. We're working on it but I sort of doubt that we will conclude it as Asian macaque. Apes also harbor P. malaria and as well as additional uncharacterized plasmodium species. What again does all this have to do with vaccine development. Well, as I showed you wild apes harbor numerous new plasmodium species one we know is the immediate precursor. What I didn't have time to tell you that 100 years ago a number of investigators actually asked the same question, can these apes parasites infect humans. They did experiments which are unthinkable today because they did blood transfusions from apes to human and from humans to apes. In the process they showed that non-laverania species which included the ape version of P. malaria and also vivax could be transmitted to humans and could -- same was true, same parasites could be transmitted from humans to apes. Laverania plasmodium falciparum did not work in the stud does. But at least for the non-laugh rainian species it was not just blood transfusion bit letting mosquitoes feed and transmit the parasites. Given magnitude of the reservoir that I just showed you the question arises, whether additional transmissions have occurred or are occurring. For vaccine development if they did he occurrence important to know what the genetic diversity is of these parasites because genetic diversities also primary challenge facing malaria vaccine development. Because ape strain would give you the outer limit of the diversity that you have to deal with. I just want to leave with you one last slide. This is a Google Earth picture, it shows a road in southeastern Cameroon, right here, and village in black that's village of Mombili the closest relative of HIV 1 group M infected chimpanzees. What you see in the red triangle are fecal samples from chimpanzees that have been collected in these locations. You can see plasmodium infected apes certainly roam within mosquitoe flying distance of human. So I think as we are going in to malaria eradication programs and vaccine development I think it is imperative to know whether these apes can be reservoir of current human infections. Lots and lots ever people that these studies, Brandon Teale in my lab ran the chimpanzee project. And others have been responsible for the malaria work. Collaborators that help us with veterinary pathologists and of course the people that have been running Gombmbe the past 40 years. Thank you. [Applause] >> Thank I nor a great talk. We'll try to reconvene in 15 minutes. If we could shoot for 3:45 that would keep us on time. Thank you. ì T&C0@T&C0@T&C TEST TEST TEST TEST TEST TEST TEST TEST TEST TEST TEST TEST TEST TEST TEST TEST TEST TEST TEST HAASE >> ALL RIGHT. I THINK WE WOULD LIKE TO GET GOING SO I'M GOING TO GIVE EVERYONE ABOUT ONE MINUTE TO GET INTO THEIR SEATS. AND THEN WE'LL GET GOING. >> ALL RIGHT. IF EVERYONE COULD TAKE THEIR SEATS MY NAME IS RICHARD CALP, I WILL BE INTRODUCING OUR NEXT SPEAKER, IT'S A GREAT PLEASURE TO INTRODUCE ASHLEY HAASE, ASHLEY IS THE HEAD OF THE DEPARTMENT OF MICROBIOLOGY AT THE UNIVERSITY OF MINNESOTA, HE HAS SPENT ABOUT THE LAST 25 YEARS OF HIS CAREER STUDYING LENTIVIRUS BOTH IN HUMAN TISSUE SAMPLES ANDSIVE RHESUS MONKEY SAMPLE AND HE'S ABLE TO DISSECT THE MECHANISMS OF HIV TRANSMISSION AND THROUGH HIS WORK WE HAVE LEARNED INCREDIBLE NEW DATA ON HOW HIV AND SIV ENTER THE HUMAN BODY AND THE MONKEY THROUGH MUCOSAL SURFACES. THIS IS GREATLY INFLUENCED OUR UNDERSTANDING OF HOW WE NEED TO DEVELOP VACCINES TO BLOCK THESE PATHWAYS. SO WITHOUT FURTHER ADIEU, ASHLEY. >> THANK YOU VERY MUCH. I'M SO GLAD TO HAVE THIS OPPORTUNITY TO CELEBRATE WITH ALL OF YOU, THE EXTRAORDINARY ACCOMPLISHMENTS AND ACHIEVEMENTS OF THIS GREAT VRC. GARY AND HIS EXCEPTIONAL CREW, I THINK THIS IS A WONDERFUL OCCASION. ACTUALLY, IT GOES BACK FARTHER THAN 25 YEARS. I'M NOW AN OLD SLOW VIROLOGIST, IT GOES BACK 35 YEARS, SOMETHING LIKE THAT. AND GARY MADE HIS FIRST OF MISTAKES LIKE HAROLD, I'M NOT GOING TO TALK ABOUT THE GUT, I'M GOING TO TALK ABOUT THE EARLIEST EVENTS THAT TRANSSPIRE AT MUCOSAL SURFACES. OF WORK IS ESSENTIALLY FOCUSED OVER THE PAST DECADE OR MORE ON THE URGENT ISSUE OF PREVENTING HIV-1 TRANSMISSION TO WOMEN THAT IS SUCH AN IMPORTANT AND CONTINUING PROBLEM IN THE PANDEMIC EPICENTER IN SUB IS A HAIR RNA AFRICA. AND WE HAVE GONE AS PEOPLE IN PATHOGENESIS DO, IN THE EARLY DAYS OF FRANK FENNER, TRYING TO USE PATHOGENESIS TO UNDERSTAND IN THIS CASE THROUGH THE FIRST ENCOUNTERS AT MUCOSAL FRONT LINES, TO PREVENT HIV-1 TRANSMISSION TO WOMEN. 1948 WITH MOUSE POX, MOST OF US THINK ABOUT PATHOGENESIS IN TERMS OF THE ANATOMIC SITES T CRITICAL EVENTS, AND THE TIMING AT WHICH THEY TAKE PLACE. SO WHEN YOU LOOK AT THOSE EVENTS, AND THESE LENTIVIRUS INFECTIONS THAT WERE NAMED O COURSE AFTER SLOW INFECTIONS IS THOSE CRITICAL EVANS OCCUR WITH INCREDIBLE RAPIDITY, WITHIN HOURS AND DAYS YOU HAVE CROSSING OF THE BARRIER, LOCAL PROPAGATION AND AS WE'LL SEE TODAY LOCAL EXPANSION THAT PLAY CRITICAL ROLES IN THE ESTABLISHMENT OF A ROBUST SYSTEMIC INFECTION. BECAUSE EVENTS TRANSPIRE WITH SUCH RAPIDITY WE MAID USE OF THE SIV RHESUS MACAQUE MODEL. IN ORDER TO UNDERSTAND THE EARLY EVENTS IN THE BOX LABELED ECLIPSE THERE. WHEN WE LOOK IN THAT BLACK BOX WHAT WE ACTUALLY SEE IS VULNERABLE OF THE VIRUS AND GREAT OPPORTUNITIES FOR PREVENTION. THOSE ARE TWOFOLD. FIRST THERE IS A SMALL INFECTED POPULATION THAT YOU HEARD A BIT ABOUT ALREADY THIS AFTERNOON. THAT EXPOSES THE VIRUS TO A VULNERABILITY AS NOT BEING ABLE TO CONTINUE THE PROPAGATION OF THAT SMALL POPULATION. I'M GOING TO PARTICULARLY FOCUS THIS AFTERNOON ON THE WHAT I CALL THE SECOND BEST CHANCE THAT'S THE NECESSITY OF THE VIRUS CONTINUING TO EXPAND LOCALLY TO ESTABLISH A SELF-PROPAGATING SYSTEMIC INFECTION. SO THIS HAS TO BE A PERIOD OF EXPANSION BEFORE THERE'S SUFFICIENT VIRUS AND INFECTED CELLS CAST ABROAD IN ORDER TO ESTABLISH A SYSTEMIC INFECTION IN THE LYMPHATIC TISSUE RESERVOIR, THAT'S OF COURSE THE FIRST TIME WE WOULD SEE AN HIV INFECTED INDIVIDUAL, REALLY TOWARD THE END OF THE SECOND WEEK AS INFECTION AT THE EARLIEST. SO WE'RE USING PATHOGENESIS HERE TO TRY TO UNDERSTAND THROUGH THE UNDERSTANDING OF PATHOGENESIS HOW WE MIGHT INTERVENE IN PREVENTING INFECTION. I'M GOING TO BE TALKING ABOUT OUTSIDE IN SIGNALING AND THE ABILITY OF A SMALL INCOMPOUND GML TO INTERFERE WITH THAT SIGNALING AND PARTICULARLY THE RECRUITMENT OF NEW TARGET CELLS THAT ARE ESSENTIAL FOR THE LOCAL EXPANSION OF INFECTION. THEN I'LL TURN MY ATTENTION TO MORE RECENT WORK THAT MAY SHARE SOME MECHANISMS WITH GML THAT'S WORK DONE WITH THE DELTA NET SIV LIVE ATTENUATED VACCINE AND PARTICULARLY THE UNUSUAL EXTRAORDINARY ACTIVITY OF TISSUE ANTIBODIES THAT WE'RE SEEING, THEIR AFFECTS AT THIS POINT OF LOCAL EXPANSION. AS MANY ARE AWARE, THIS IS A HIGH DOSE MODEL OF HIV-1 TRANSMISSION IN FEMALE RHESUS MACAQUES. WE INOCULATE AND EXPOSE THEM TO OVER 200,000 INFECTIOUS DOSES OF VIRUS NOT BECAUSE WE NEED A CONSISTENT INFECTION BUT BECAUSE WE WANT A RIGOROUS SET OF CHALLENGES AND FROM MY PERSPECTIVE LOOKING AT TISSUES AND TISSUE ANALYSIS I NEED AN INFECTION ON A NEED TO HAVE ENOUGH POSITIVE EVENTS WHEN WE SAMPLE A TISSUE TO SEE KEY EVENTS IN THE VIRUS HOST INTERACTIONS THAT WILL FULFILL THE CRITERION OF DIRECTLY OBSERVED PATHOGENESIS. DESPITE THE FACT WE EXPOSE THE ENVIRONMENT TO A HUGE DOSE OF VIRUS, THE CLOSE TO 10 BILLION PARTICLES THIS IS AT THE MICROSCOPIC LEVELS AND WE FIND MOST VIRUS IN THE CASE IN THE MUCUS, TRAPPED IN THAT MUCOUS. WHETHER IT'S A RESULT OF THAT TRAPPING MECHANISM OR DELUSION THE OUTCOME IS TARGET CELLS IN THE SUB MUCOSAL ARE EXPOSED TO A RELATIVELY SMALL AMOUNT OF VIRUS. THE OUTCOME OF THAT IS THAT WE ONLY HAVE INFECTED SMALL FOUNDER FOP -- POPULATIONS. IF YOU'RE OPERATIONALLY DEFINED THE PRESENCE OF INFECTION AT THE SITE OF ENTRY BY RNA IN SITU HYBRIDIZATION, WE SEE OUR SIGNAL AT 3 TO 4 DAYS. IN THE REGION OF THE ENDOCERVIX. SO IF YOU LOOK HERE AT THE CERVIX, WE HAVE THE ENDOCERVIX, THE TRANSFORMATION ZONE AND THE ECTO CERVIX AND WHEN WE SCAN 25 TO 50 SECTIONS OR SO, WE USUALLY ACTUALLY INVARIABLY FIND A SMALL FOCUS AS SHOWN HERE ENCIRCLED IN RED. THIS IS ONE OUT OF 40 SECTIONS, AND IS A MONTAGE OF ANOTHER 40, 10X IMAGES CREATE HEERD TO GIVE YOU AN IDEA OF THE SIZE OF THAT ORIGINAL FOUNDER POPULATION, FOUR DAYS AFTER EXPOSURE. SO THAT IS THE FIRST BEST CHANCE, YOU HAVE A SMALL INFECTED FOUNDER POPULATION, YOU HAVE HEARD ABOUT THE GENETIC EFFECTS THAT YOU ALSO GET AT THAT POINT. THAT CERTAINLY IS AN OPPORTUNITY FOR SOME OF THE EARLIEST INTERVENTIONS, TO PREVENT THE ESTABLISHMENT ALL TOGETHER OR TO CONTAIN IT AT THAT STAGE. BUT THERE IS A SECOND CHANCE. THAT HAS TO DO WITH THE FACT THAT THE VIRUS NEEDS TO EXPAND THAT LOCAL INFECTION IN ORDER TO ESTABLISH A ROBUST SYSTEMIC INFECTION. WITHOUT THAT ANTECEDENT EXPANSION WE DON'T FIND EVIDENCE OF A ROBUST SYSTEMIC INFECTION. WE MIGHT MAY FIND EVIDENCE OF OCCULT INFECTIONS BUT TO GET THE SYSTEMIC INFECTION THROUGHOUT THE LYMPHATIC TISSUE WE NEED LOCAL EXPANSION. JUST LOOKING AT WHAT THAT LOOKS LIKE, MAPPING THE POSITIONS AND NUMBERS OF THE INFECTED CELLS ON A TWO DIMENSIONAL GRID HERE AT FOUR DAYS IS MAP OF THAT ON A TWO DIMENSIONAL GRID. THREE DAYS LATER WHAT YOU SEE IS NOT A DIFFUSE EXPANSION OF MULTIPLE SITES OR A DIFFUSE EXPANSION LIKE RIPPLES FROM A SMALL ROCK THROWN INTO A POND, YOU SEE THE SAME CLUSTERING BUT NOW ON A MUCH LARGER DIMENSION THAT'S CONTINUED TO GO BY THE CO-LESS SENSE AND ACTUALLY CLOSE ENCOUNTERS OR CELL TO CELL SPREAD TRANSMISSION OF THE VIRUS FROM ONE CELL TO THE NEXT. MAINLY AT THESE SITES OF INITIAL FOUNDER POPULATIONS. IF WE NOW LOOK A MERE THREE DAYS LATER THIS IS A TEN DAYS POST INNOCULATION AND I'M ILLUSTRATING TWO THEMES. ONE CLUSTER OF INFECTED CRELINGS, THE OTHER UNDERLYING AREAS OF INFLAMMATION. THIS IS THE TRANSFORMATION ZONE BETWEEN THE ENDOAND ECTO CERVIX, THE COALESCE SENT GREENISH MASS CONSIST OF THOUSANDS OF INFECTED CELLS. YOU CAN SEE THE ACCRETION AROUND THE INITIAL FOCI AS WELL. THESE DARK SPANNING NUCLEI REPRESENTING IMMENSESIVE INFLAMMATORY INFILTRATE WE CAN DETECT AT THAT TIME. NOT ONLY DO WE SEE ACCRETION AROUND THE OTHER CLUSTER BUT THE SERPENTINE ARROW PREVENT IT IS SPREAD OF INFECTION ALONG AN INFLAMMATORY TRACT OR TRACT OF INFLAMMATORY CELLS THAT HAVE COME IN SO THE VIRUS IS GOING ALONG AS IT FINDS SUB TRAITS TO CONTINUE TO REPLICATE AND SPREAD. JUST TO SUMMARIZE THAT POINT, WE GO AT FOUR DAYS FROM THAT SMALL CLUSTER. MAPPED HERE THREE DAYS LATER IT HAS THESE DIMENSIONS AN THREE DAYS LATER HAS THESE EXPANSIONS, BOTH FOR THE INITIAL INFECTED FOUNDER POPULATION AS WELL AS UNDERLYING INFLAMMATION. YOU MAY ASK AND WE CERTAINLY DID, IF YOU CONSIDER THE INITIAL BOUNDARY CONDITIONS THE VIRUS CONFRONTS WHEN IT COMES ACROSS A MUCOSAL SURFACE, PARTICULARLY IN THE CERVIX, IT ENCOUNTERS SCATTERED LYMPHOCYTES WITH CD4 T-CELLS THE MAJOR HOST IN THIS INITIAL PROPAGATION AND EXPANSION. THESE CELLS TURNED OUT TO BE QUITE DISPERSED AND LOW DENSITY. SO IF THE VIRUS IS SUCCESSFUL IN ESTABLISHING THE FOUNDER COPULATION IT CON CONFRONT IT IS PROBLEM HOW TO EXPAND. THE INFECTION GOING A FEW DAYS PORT OF ENTRY FROM FEW TO MANY CELLS IS A CONSEQUENCE OF A SUPPLY OF NEW TARGET CELLS. SO IF YOU LOOK AT THE DENSITY OF THE POPULATIONS OF THE SUBSTRATES WHICH THE VIRUS IS GOING TO REPLICATE SAY IN BROWN HERE AND OTHER INFECTED ANIMAL YOU SEE A FEW EPITHELIAL LEUKOCYTES IN THE MUCOSEA AND THERE'S NO INCREASE BUT THERE'S A REMARKABLE INTERCHANGE BY THE FOUR DAYS POST INNOCULATION, YOU CAN SEE ALL OF THESE BROWN STAIN CELLS JUST UNDERNEATH THE EPITHELIAL BARRIER AND AT SIX DAYS IT'S MORE DRAMATIC. SO WHAT'S HAPPENED HERE IS THE VIRUS SOMEHOW IN INITIAL INTERACTIONS WITH ITS HOST AN MUCOSAL SURFACES HAS CHANGED THE LANDSCAPE. SO YOU HAVE A LANDSCAPE HERE CD4 POSITIVE T-CELLS IN BROWN SHOWN AT HIGHER MAGNIFICATION UNDER THE EPITHELIAL SURFACES YOU CAN SEE THE COLLECTIONS, LARGE COLLECTIONS OF BROWN STAINING CD4 POSITIVE T-CELLS. SO JUST AT THE TIME THE VIRUS IS BASICALLY BURNED THROUGH ITS INITIAL CONCENTRATION OF SUBSTRATES, IT NOW HAS NEW TARGET CELLS TO INFECT. AND YOU CAN SATISFY AN IMPORTANT PREDICTION THAT THOSE ARE TARGETS ALSO THE ONES THAT ARE INFECTED BY COMBINING IN SITU HYBRIDIZATION TO DETECT VIRUS RNA WITH IMMUNOHISTOCHEMISTRY TO DETECT T-CELLS IN THIS CASE. ESSENTIALLY 90 TO 95% OF CELLS INFECTED AFTER DAY FOUR ARE THE SAME CD4 T-CELLS WITH WHICH INFECTION ACTUALLY BEGAN. WELL, IF THE ANSWER TO HOW THE VIRUS GOES FROM THE SMALL POPULATION AT THE PORT OF ENTRY TO SYSTEMIC INFECTION OVER A FEW DAYS IS THE SUPPLY OF NEW TARGET CELLS THE NEXT LOGICAL QUESTION IS, WHAT INDUCES THAT INFLUX OF NEW TARGET CELLS? THE ANSWER IS TURNED OUT TO BE THE INNATE IMMUNE RESPONSE TO INFECTION OR EXPOSURE. INITIATED AT THE MUCOSAL EPITHELIAL THE TRUE FRONT LINE OF THE IMMUNE SYSTEM. AND IT TURNS OUT TO INVOLVE CIRCUITRY WE CALL OUTSIDE IN SIGNALING. EXPOSURE TO THE VIRUS IS SOMETHING IN THE INOCULUM ELICIT IT IS PRODUCTION INCREASED PRODUCTION OF MITT 3 ALPHA OR CCL-20 IN THE EPITHELIUM. THAT IN TURN ATTRACTS CCI OF 6 POSITIVE DENDRITIC CELLS THAT ELABORATE 1 BETA AND OTHER CHEMOKINES AN THOSE CHEMOKINES BRING IN THE NEW TARGET CELL THAT ALLOWS THE GROWTH AND SPREAD FROM THAT INITIAL CLUSTER THROUGH ACCRETION OF NEW TARGET CELLS. WE GOT ON BY A VARIETY OF DIFFERENT APPROACH, ONE APPROACH IS TO SIMPLY COMPARE UNINFECTED ANIMAL WITH A TRANSCRIPTSAL PROFILE WHAT HAPPENS IN LOCAL INFECTION WITHIN THE FIRST FEW DAYS OF INFECTION. AND WHEN WE LOOK THROUGH THE GENE WHAT WE SAW AT THE TOP OF THE LIST AT DAY 1 AND 3 POST INNOCULATION IS A 4 AND 5 FOLD INCREASE IN EXPRESSION OF MITT 3 ALPHA. AND WHEN WE LOOK FOR EXPRESSION OF PROTEIN WE INDEED FOUND THE PROTEIN BEING EXPRESSED AND THE ENDOCERVICAL EPITHELIUM. THE NEXT PART OF THE CIRCUITRY, INVOLVES THE PLASMA OR THE PDCs, THAT WAS A SEPARATE AND REMARKABLE OBSERVATION MADE BY JAKE ESTES A POST-DOC IN MY LAB. HE WAS TRYING TO UNDERSTAND A PREMATURE T REGULATORY RESPONSE AT THE PORTAL OF ENTRY. AND STAINING FOR PDCs SHOWN HERE IN BROWN WHAT HE FOUND WAS REMARKABLE LINING OF PDCs JUST UNDERNEATH THE MUCOSAL EPITHELIUM. SO THESE CELLS OF COURSE ARE THE CHAMP PRODUCERS OF INTERFERON, IT IS SHOWN HERE IN FACT PRODUCING LARGE AMOUNTS OF INTERFERON ALPHA. SO THEY'RE DOING THE KINDS OF THINGIOUS EXPECT INNATE SYSTEM TO BE DOING TO PREVENT INFECTION AND ITS EXPANSION. BUT WHETHER IT INVOLVES SOME MECHANISM FOR THE RESISTANCE OF CD4 T-CELLS INTERFERON, WHILE INTERFERON MAY PROTECT V CELLS IT DOESN'T PROTECT THE CD4 T-CELLS IN THAT ENVIRONMENT. THE BALANCE HERE, OUR BALANCE IS WE HAVE THIS FAVORABLE ENVIRONMENT OF NEW TARGET CELLS WHICH THE VIRUS IS ABLE TO EXPAND. SO PARADOXICALLY AND COUNTER INTUITIVE THE HOST INNATE IMMUNE RESPONSE IS PLAYING A CRITICAL ROLE IN FACILITATING TRANSMISSION. THAT WAS THE RATIONALE BEHIND THE COUNTER INTUITIVE STRATEGY WE CAME UP WITH A FEW YEARS AGO. IF WE COULD INHIBIT OR MODERATE THAT INNATE AND INFLAMMATORY RESPONSE AT THE PORTAL OF ENTRY, WE MIGHT THERE BY BE ABLE TO PREVENT INFECTION. SO WE'RE LOOKING AGAIN, BETWEEN THE LINKAGES BETWEEN INNATE IMMUNITY AND INFLAMMATION AN TARGET CELL AVAILABILITY AS PROOF OF PRINCIPLE, COULD WE TARGET THE TARGET CELL AVAILABILITY, THE MUCOSAL SIGNALING AND THE INNATE IMMUNE RESPONSE AND INFLAMMATION AND THERE BY ACTUALLY PREVENT ACUTE SYSTEMIC INFECTION. WE CHOSE TO DO THAT WITH A COMPOUND GML GLYCEROL MONOLIGHT SHOWN HERE IN THE CARTOON P IS TIPPING THE BALANCE IN THE FAVOR OF THE HOST BUT NOT ONLY RESTRICTING INFECTION BUT RESTRICTING ACCESS TO TARGET CELLS FOR THAT EXPANSION. WHY GML? GML IS A FATTY ACID MYLOESTER IN HUMAN BLOOD AND MILK. IT'S GENERALLY RECOGNIZED A SAFE AGENT BY THE FDA. THE REASON WE GOT IT IS ONE FACULTY MEMBER IN MY DEPARTMENT HAD BEEN STUDYING IT FOR NEARLY TWO DECADES NOW AND HAD SHOWN BACK IN 1992 THAT IT WOULD HAVE A T-CELL ACTIVATION AN TOXIC SHOCK SYNDROME TOXIN ONE. SO PAT IN MY DEPARTMENT AND I HEAR HIM TALK AT EVERY RETREAT. IF WE COULD INHIBIT T-CELL ACTIVATION THAT WE MIGHT BE ABLE TO PREVENT SIV REPLICATION. WE SHOW IN VITRO THAT WE CAN IN FACT INHIBIT BY LIMITING THE ACTIVATED PBMCs WE COULD INHIBIT REPLICATION OF HIV-1, MORE PERTINENT WE COULD SHOW WITH EPITHELIAL CELL CULTURES IF WE ADDED HIV WE WOULD GET AN EXTENSIVE BUMP IN MITT 3 ALPHA EXPRESSION AN INHIBIT WITH GML. THEN WE DISCOVERED MORE PERTINENTLY IN THE MONKEY SYSTEM WE FOUND THAT WITH A WASHING PERIOD OF ABOUT FOUR OR FIVE DAYS, WE COULD GREATLY REDUCE THE AMOUNT OF MITT 3 ALPHA PRESENT IN CERVICAL VAGINAL FLUIDS. SO WE DID EXTENSIVE SAFETY STUDIES OVER A 3 TO 6 MONTH PERIOD FOR DAILY EXPOSURE. WE WERE ABLE TO SHOW A COUPLE OF YEARS AGO IN THE INITIAL PILOT EXPERIMENTS THAT WE COULD PROTECT FIVE OUT OF FIVE MONKEYS AGAINST SYSTEMIC INFECTION COMPARED AGAINST ALL THE CONTROLS. AND WE'VE GONE ON AT THIS DEMONSTRATION THAT GML HAD POTENTIAL NOT ONLY BECAUSE IT WAS SAFE AND EFFICACIOUS, IT'S ALSO QUITE CHEAP, LESS THAN A PENNY PER APPLICATION AND WAS EFFICACIOUS. THE MORE RECENT WELL POWERED EFFICACY TRIALS WE HAVE BEEN DOING IN 15 GML TREATED ANIMALS, 15 HAVE BEEN FAIRLY PROTECTED AGAINST ACUTE REPLICATION OF SIV ROBUST SYSTEMIC INFECTION VERSUS 415 NOT OF THE THE CONTROLS WITH A VERY SIGNIFICANT P VALUE. SO I WANT TO TURN NOW FROM WHAT WE'VE -- I HAVE BEEN SHOWING AND TELLING YOU AB OUTSIDE IN SIGNALING THE IMPORTANCE OF TARGET CELLS AND EXPANSION FOR ROBUST SYSTEMIC INFECTION AND THE ABILITY OF GML THROUGH BLOCKING OR MODERATING THAT SIGNALING TO ACTUALLY PREVENT EXPANSION NECESSARY FOR ROBUST SYSTEMIC INFECTION. AND TURN TO MORE RECENT WORK WE HAVE BEEN DOING ON DELTA NEFF LIVE ATTENUATED VACCINE. THIS HAS A VERY LONG HISTORY AS YOU KNOW, ALMOST AGAIN TWO DECADES OLD FROM THE ORIGINAL DEMONSTRATION OF THE PROTECTIVE EFFECTS OF PRIOR INFECTION. IV INFECTION WITH LIVE ATTENUATED SIV VACCINE FROM ROD ROIS'S GROUP IN 1992. AS YOU KNOW THERE WERE SAFETY ISSUES THAT PRECLUDED ADVANCING THIS TO HUMAN TRIALS BUT IT CERTAINLY SEEMS AND CONTINUES I THINK TO BE IMPORTANT TO IDENTIFY CORRELATES OF THAT PROTECTION THAT MIGHT ENABLE THE DESIGN OF AN EFFECTIVE HIV-1 VACCINE. WE HAVE BEEN STUDYING THE VACCINE IN CHALLENGE PHASES IN COLLABORATION WITH PAUL JOHNSON. ANOTHER REMARKABLE FACT ABOUT THIS PROTECTION IS THE MATURATION OF PROTECTION. SO IF YOU INFECT WITH DELTA NEFF IV YOU LOOK AT WEEK 5, THE ABSOLUTE PEAK OF THE CELL IMMEDIATE WRAITED -- MEDIATED IMMUNE RESPONSE THERE'S NO INFECTION. IF YOU WAIT UNTIL WEEK 20 OR 40, QUITE STRIKING STERILELIZING OR ATTENUATION. SO WE HAVE BEEN SEEKING CORRELATES OF THAT MATURATION OF PROTECTION TO ENABLE THE DESIGN AND TESTING OF VACCINE CANDIDATES. THIS SHOWS SOME OF THE FIGURES ON THE LEVEL OF PROTECTION HERE, HERE ARE THE WILE TYPE ANIMALS. SOME PEAK REPLICATION IN THE CASE OF A CHALLENGE AT FIVE WEEKS BUT YOU GET HIGHLY SIGNIFICANT DECREASES BOTH IN PEAK REPLICATION AND IN SET POINT REPLICATION IN STUDIES THAT PAUL IS DOING WITH A HIVRAD GRANT. WHEN WE LOOK FOR CORRELATES IN THIS MODEL, WE HAVE BEEN LOOKING AT ALL THE SITES, THE LOCAL, THE PORTAL OF ENTRY AS WELL AS THE SYSTEMIC SITES, THE DRAINING LYMPH NODES AND A VARIETY OF SYSTEMIC SITES, I SHOWED YOU EARLIER IN THIS TALK THERE'S A REMARKABLE SMALL FOUNDER POPULATION OR REMARKABLE EXPANSION OF THAT POPULATION OVER A VERY BRIEF PERIOD FROM DAYS 4 TO ABOUT 10. IF YOU LOOK AT THE CONCENTRATION OF VIRUS PRESENT, IN THESE TISSUES BY DAY FOUR IT'S DETECTABLE, IT'S CLOSE TO A MILLION COPIES PER MICROGRAM OF IT SHALL SHOE RNA TO WELL OVER THAT IN THIS BRIEF TIME FRAME. A REVIEW OF THAT AGAIN, I SHOWED YOU THE SMALL FOCUS THAT'S DETECTABLE AT DAY FOUR. SHOWED YOU THE EXPANSION OF THE TARGET CELL POPULATION AT DAY 4 TO 6 AND THEN THIS TREMENDOUS AMPLIFICATION AND SYSTEMIC INFECTION. THE MOST STRIKING THING WE SAW IN THESE ANIMALS STRAIGHT AWAY WERE THEY WERE STILL INFECT SOD HERE IS THAT SAME CERVICAL CLUSTER THE GREEN CELLS, A FOCUS YOU HAVE TO LOOK HARD FOR BECAUSE THE SAME SMALL FOUNDER POPULATIONS, BUT IN CONTRAST TO THE EXPANSION OF THAT FOUNDER POPULATION, LOOKING DOWN AT DAY 11 ESSENTIALLY THERE'S BEEN NO EXPANSION. THAT'S ASSOCIATED WITH THE FACT THERE'S NO RECRUITMENT OF TARGET CELLS SO THE BOTTOM OF THE RARE BROWN CELLS YOU CAN SEE HERE ARE CD4 T-CELLS, THIS IS ACTUALLY PRESENT I THINK AT 11 DAYS POST EXPOSURE. SO IN CONTRAST TO THAT HUGE INFLUX OF TARGET CELLS IN THIS CASE WE DON'T HAVE ANY INFLUX AT ALL. SO THE FIRST CORRELATE IS MO A SMALL FOUNDER POPULATION BUT NOLOCAL EXPANSION AND THERE'S A HUGE INCREASE IN SYSTEMIC VIRAL LOADS. SIX ORDERS OF MAGNITUDE IN THE CASE T CERVICAL TISSUES AND COMPARABLE 3 TO 4 ORDER OF MAGNITUDE OUT IN THE SECOND WEEK OF INFECTION DURING A HUGE SYSTEMIC INFECTION IN THE CASE OF AS I SAY THE LYMPHATIC TISSUES. SO WE ESTABLISH INFECTION, NO INCREASE IN TARGET CELL, NO EXPANSION, THAT'S ASSOCIATED WITH A QUITE STRIKING DECREASE IN THE LOCAL EXPANSION OF SYSTEMIC INFECTION. I'LL COME BACK TO THIS AT THE VERY END. MY BIAS IS WHAT TURNED OUT TO BE WHAT I CALLED ENOUGH AND SOON ENOUGH IN TERMS OF A T-CELL RESPONSE AT THE PORTAL OF ENTRY, RESIDENT POPULATION OR A RAPID RESPONDER POPULATION THAT WOULD CONTAIN INITIAL FOUNDER POPULATION. THERE IS NO DIFFERENCE IN 5 AND 20 WEEKS OF VIRUS CD8 SPECIFIC T-CELLS. THERE'S NO EXPANSION OF THOSE CELLS. AND THE ONLY CORRELATE WE HAVE FOUND SO FAR WITH THIS DRAMATIC EFFECT ON THE LOCAL EXPANSION ARE TISSUE ANTIBODIES TO ENVELOPE THAT ARE ACTUALLY BEING PRODUCED BY PLASMA CELLS AT MUCOSAL SURFACES SO THAT YOU HAVE A CONCENTRATED ANTIBODY RESPONSE ENTIRELY FOCUSED AT THAT PORTAL OF ENTRY. SO THE OBSERVATION NOW IN FULL ANIMALS AT 5 AND 20 WEEKS AT THE COMPARISON OF IGG AND IGA LEVELS YOU CAN SEE THE ANTIBODIES MAKE THE ANIMALS MAKE ANTIBODIES AGAINST THE GATE PROTEIN. SOME INCREASES AT 20 WEEKS BUT THE STRIKING CHANGE ARE THESE TWO ENVELOPE BANDS PARTICULARLY TO GP-160, IGG GREATER THAN IGA. THE DIFFERENCES RANGED AS MUCH AS 12 FOLD TO 170 FOLD. SO QUITE DRAMATIC INCREASES IN THE AMOUNT OF ANTIBODY. WHEN (INDISCERNIBLE) WAS A GRADUATE STUDENT IN MY LABORATORY LOOKING FOR IGG POSITIVE CELLS HE FOUND CELLS SHOWN HERE, DIFFICULT TO SEE I THINK, AT FIVE WEEKS BUT QUITE DRAMATIC INCREASES IN CERVIX AT 20 WEEKS. LET ME SHOW YOU THAT A LITTLE HIGHER MAGNIFICATION, IT'S QUITE STRIKING. A CIRCLE OF RED PLASMA CELLS, IGG POSITIVE CELL, WHAT YOU SEE HERE, THIS IS THE LUMIN, THIS IS THE ENDOCERVICAL EPITHELIUM AND LINED UP UNDERNEATH ABSOLUTELY IN A CONTINUOUS LINE ARE PLASMA CELLS. THE PLASMA CELLS NOT ONLY BECAUSE THEY LOOK LIKE PLASMA CELLS, IN FACT THEY HAVE THE MARKERS FOR PLASMA CELLS, CD 27 AND CD 138, AND THE MERGED IMAGES HERE YOU CAN SEE THE CELLS LINING UP UNDERNEATH THE EPITHELIUM BOTH FOR CD 27 FOR CD 138. SO WHAT I HAVE TOLD YOU ABOUT TODAY ARE TWO INTERVENTIONS BASED ON PATHOGENESIS UNDERSTANDING WHAT HAPPENS THROUGH FIRST ENCOUNTER AS T AT MUCOSAL SURFACES OF ONE IS SIGNALING OF TARGET CELLS AND EXPANSION THAT GML INTERFERES WITH AND HAS EFFECTS ON THE SUBSEQUENT CORES AN SYSTEMIC INFECTION. THERE ARE PARALLELS AS WELL IN TERMS OF MATURATION OF THE PROTECTIVE RESPONSE TO THAT LIVE ATTENUATED VACCINE, THAT THESE TISSUE ANTIBODIES MAY INTERFERE WITH THE EXPANSION RECRUITMENT AND EXPANSION. PROBABLY MOST IMPORTANTLY LEAVE US WITH TWO VERY CONTRASTING IMAGES OF WHAT GOES ON IN I.E. VERSUS THE VACCINATED ANIMALS. I'M SHOWING YOU THE IMAGE FROM JAKE'S WORK, THE PDC IS LINING UP IN INNATE RESPONSE THAT FA UNTIL SATE SYSTEMIC INFECTION AND I SHOW HERE THE LINEUP OF PLASMA CELLS THAT TAKES PLACE TO CONCENTRATE THE PRODUCTION OF THE ANTIBODY AT THE PORTAL OF ENTRY THAT WE THINK HAS PROFOUND EFFECTS ON LOCAL EXPANSION. AND THE SUBSEQUENT COURSE OF INFECTION. I BORROWED THIS SLIDE FROM PAUL JOHNSON WHO IS LOOKING FOR CAUSE PROTECTION. PAUL REPOSITIONED ANATOMICALLY WITH A PARTICULAR PERSPECTIVE ON THE SOON ENOUGH PERSPECTIVE ON CD POSITIVE T-CELLS. SO I'VE ALTERED THAT PERSPECTIVE SLIGHTLY. NOT THAT I HAVE ABANDONED THE PERSPECTIVE BUT MOVED TO THE FRONT OF THE ANIMAL WHERE THE ANTIBODIES MIGHT BE QUITE IMPORTANT IN THAT PO PROTECTION. LET ME END WITH THE CREDITS AND ACKNOWLEDGE MS FOR MY LABORATORY. AT THE UNIVERSITY OF MINNESOTA. PARTICULARLY THE WORK OF CHIN SING LEE, MING XONG JASMINE DUWON. PAT SLEVERT, MANDY, A GRADUATE STUDENT IN HIS LABORATORY, THE PRIMATE RESEARCH CENTER CREDITS FOR THE WORK ON GML. AND FOR THE WORK ON THE DELTA NEFF ATTENUATED VACCINE, THAT'S DONE WITH THE SUPPORT OF PAUL JOHNSON AND KEITH RES, WE HAVE HAD CONTINUING HELP FROM JAKE ESSI AND JEFF LIFSON AT THE NATIONAL CANCER INSTITUTE. AND I THANK YOU. [APPLAUSE] >> I'M IMARNY GRAHAM FROM THE VRC, IT'S MY PLEASURE TO INTRODUCE DR. GLENDA GRAY. SHE'S TRAVELED THE FARTHEST FROM JOE HANSBURG, SOUTH AFRICA. SHE'S PROFESSOR OF PEDIATRICS AT THE UNIVERSITY OF (INDISCERNIBLE) IN JOHANNASBURG. ON THE TOP FLOOR LOOKING OUT OVER THE SUETTO TOWNSHIP. FROM THAT POSITION SHE'S ABLE TO KEEP HER FINGER ON THE PULSE OF THIS HIV EPIDEMIC AND I DON'T THINK THERE'S ANYONE IN THE WORLD IS BETTER ABLE TO DESCRIBE TO US THE REASON OUR WORK IS NOT FINISHED UNTIL WE HAVE AN HIV VACCINE. SHE HAS BEEN AWARDED SEVERAL HONORS, SHE GOT THE WOMAN OF THE '90s AWARD FROM VAMINA FOR HER WORK ON PERINATAL HIV RESEARCH. AND A VERY DISTINGUISHED AWARD, NELSON MANDELA HEALTH AND HUMAN RIGHTS AWOR FOR WORK ON MOTHER -- AWARD FOR MOTHER TO CHILD TRANSMISSION. IN 2000 SHE'S BEEN INVOLVED IN VACCINE DEVELOPMENT, SHE'S DIRECTOR OF THE AFRICAN COMPONENT OF THE VACCINE TRIALS NETWORK AND SHE'S BEEN A GREAT SPOKESPERSON FOR VACCINES SINCE SHE CAME IN TO THIS FEEL. SO GLENDA. [APPLAUSE] >> THANKS, BONNIE FOR A WONDERFUL INTRODUCTION AND THANK YOU, GARY FOR INVITING ME TO DO THIS TALK. AND CONGRATULATIONS OF TEN YEARS OF MAGNIFICENT WORK AND YOU SHOULD BE VERY PROUD OF ALL THE THINGS THAT YOU HAVE ACHIEVED AT THE VRC. I'M GOING TO TALK, AS BONNIE SAID I'M GOING TO CONTEXTUALIZE THE REASON HIV VACCINE IS IMPORTANT AND SHOW YOU SOME OF THE BURDEN OF HIV AN WHAT HAPPENS AT A POPULATION LEVEL WHEN HIV TAKES ROOT. SO I'LL START MY TALK SAYING UNFORTUNATELY HIV IS NOT ANYTHING SPECIAL IN SOUTH AFRICA AND INTERVENTIONS WILL HAVE TO BE BOLL AND MULTI-FACETTED. SO WE HAD A POPULATION OF 49 MILLION PEOPLE, OUR LIFE EXPECTANCY IS ONLY 49 YEARS. WE HAVE A PREVALENCE OF 10.9% WHICH MEANS ONE IN TEN SOUTH AFTER CANS ARE HIV INFECTED. WE HAVE AN UNENVIABLE POSITION OF BEING THE COUNTRY WITH THE LARGEST NUMBER OF PEOPLE INFECTED WITH HIV, AND THE LARGEST INTEREST OF OUR PROGRAM IN THE WORLD, REPRESENTS ABOUT 30% OF PEOPLE WHO REQUIRE (INAUDIBLE) TREATMENT. TO UNDERSTAND THE EPIDEMIC IN SOUTH AFRICA WE NEED TO STEP BACK AND SEE WHAT HAPPENED AND WHY DID THE EPIDEMIC TAKE ROOT IN SOUTH AFRICA. AT THE SAME TIME AS WE WITH MANDELA WITH THE FIRST ELECTION IN THE WAY ONE CONVERTS THEY WOULD -- (INAUDIBLE) YOU CAN SEE AT THE BOTTOM SLIDE HERE WITH NOT GOOD AT POINTERS BUT I'LL EXPLAIN IT. YOU CAN SEE THAT WITH THAT TRANSMISSION TO DEMOCRACY, AT THE SAME TIME SOMETHING TERRIBLE WAS TAKING ROOT. SO SOMETHING TERRIBLE WAS TAKING ROOT AT THE SAME TIME AS THE DEMOCRACY WAS OCCURING IN SOUTH AFRICA. AND UNDER MANDELA WORK HAD TO BE DONE. WE HAD TO RELOOK AT EDUCATION, HEALTH AND GOVERNMENT AND SO THERE'S A VERY LITTLE TIME TO PAY ATTENTION TO A EPIDEMIC THAT WAS TAKING ROOT. AND AFTER THE SECOND ADMINISTRATION OF THE AMC (INDISCERNIBLE) OFFICE AND HIS OFFICE WAS -- BY ASSOCIATIONS WITH (INDISCERNIBLE) AND THE EARLY RESPONSE IN THE FIRST TEN YEARS POST DEMOCRACY WAS SLOW AND ONE OF THE REASONS WERE WE SAW SUCH AN INCREASE IN HIV IN SOUTH AFRICA. THE FIRST TIME SOUTH AFRICA THOUGHT ABOUT THE CHANGES IN THE DEATH RATES WAS AT THE TIME OF THE PRESIDENTIAL -- CONVENED THE PRESIDENTIAL AIDS COUNCIL IN 2000 TO LOOK AT WHETHER HIV CAUSED AIDS. AND THE COUNCIL CONVENE AD STUDY TO LOOK AT THE RATE OF DEATH IN THE COUNTRY. I SAW SOMETHING ALARMING THAT THE RATE AND THE PROFILE HAVE (INAUDIBLE) IN SOUTH AFRICA. SO WHAT YOU SAW WAS INCREASING DEATH BY ABOUT 90% IN A 8 YEAR PERIOD. PARTICULARLY IN THE AGE 25 TO 49. INCREASE OF 173%. AN THIS WAS A POPULATION BASED APPROACH TO SHOW THAT SOMETHING WAS HAPPENING IN SOUTH AFRICA. AND THE ONLY THING THAT COULD ACCOUNT FOR IT WAS INCREASE IN HIV IN SOUTH AFRICA. THERE'S MORE (INDISCERNIBLE) WAS INCREASE IN THE NUMBER OF FEMALE DEATHS IN SOUTH AFRICA. AND THE WOMAN IN THE EARLY REPRODUCTIVE AGE GROUP OUTSTRIPPED ALL THE WOMEN IN THE AMOUNT OF DEATH. IF YOU DON'T HAVE A WOMAN IN THE HOUSEHOLD, THIS IMPACTS CHILD SURVIVAL AND INCREASES HOUSEHOLD VULNERABILITY SO THE INCREASE IN FEMALE DEATHS IN THE PRIME OF THEIR LIFE WAS INCREDIBLY DEVASTATING TO THE FABRIC OF SOCIETY IN SOUTH AFRICA. >> WHAT IS HIV RIS INCOME SOUTH AFRICA? WE CAN MEASURE IT. WE CAN LOOK AT HIV PREVALENCE AND INCIDENCE AN DESCRIBE THE PROFILE OF HIV WE HAVE SEEN IN SOUTH AFRICA. WE SEE THREE RISKS, ONE IN THE MOTHER TO CHILD PEDIATRIC HIV FROM MOTHER TO CHILD TRANSMISSION FROM HIV WOMAN, HIV INFECTED WOMEN WHEN PREGNANT. WE SEE A HUGE RISK OF HIV ACQUISITION BY SEXUAL TRANSMISSION IN EARLY ADULTHOOD AND THEN WE START TO SEE IN OLDER WOMAN AN EMERGING EPIDEMIC WHICH IS DESCRIBED AND HASN'T REALLY BEEN INVESTIGATED MUCH. IF YOU HAVE A LOOK AT THE YOU ARE VAI AT THE KNOWLEDGE AND BEHAVIOR OF SOUTH AFRICANS OVER TIME IT'S IMPORTANT TO UNDERSTAND WHAT POPULATION IN SOUTH AFRICA THINK AND BELIEVE ABOUT HIV AND WHAT ARE THE ACTIONS TO TRY AND LOOK AT INTERVENTIONS. AND THIS SHOWS A COMMUNICATION SURVEY, WITH KNOWLEDGE AND BEHAVIOR. YOU SEE ONLY 85% OF PEOPLE IN 2009 BELIEVE CONDOM USE COULD PREVENT HIV. ONLY 40% USED CONDEMNS TO PREVENT AIDS AND ONLY IN A SIMILAR AMOUNT, 30% AT THE LAST (INAUDIBLE). THE GOOD NEWS PEOPLE ARE GETTING HIV TESTED IN SOUTH AFRICA DEMAND A SURVEY, 61% OF PARTICIPANTS SURVEYED ACTUALLY HAD AN HIV TEST AND HALF OF THOSE HAD ANYTIME THE LAST 12 MONTHS. STILL A LOT OF MULTIPLE PARTNERS. ONE IN FIVE MEN IN SOUTH AFRICA REPORT HAVING MULTIPLE PARTNERS. SO I HAVE MENTIONED, I HAVE TRIED TO QUANTIFY WHAT HIV RISK IS. AND IT'S HARD TO SIGH HOW TO MEASURE HIV RISK BUT WE DON'T UNDERSTAND WHAT IS HIV RISK. IN A HIGH PREVALENT SOCIETY WITH HIGH RATES OF HIV INFECTION IN HOUSEHOLDS, PEOPLE ARE GRAPPLING WITH LONG TERM INFECTIONS, AND FEAR OF TRANSMISSION. AND THERE'S A GREAT DEAL OF INSECURITY IN LONG TERM RELATIONSHIP, MOST IN SOUTH AFRICA ARE NOT MARRIED AND THEY MAY SPEND THREE OR FOUR TIME AS WEEK WITH THEIR PARTNERS BUT THESE AREN'T LONG TERM IN HOUSE RELATIONSHIPS. SO THERE'S A GREAT DEAL OF INSECURITY AND OFTEN INDIVIDUAL VERSUS TO BALANCE LOVING AND HAVING A RELATIONSHIP WITH SOMEONE WHILE TRYING TO PREVENT THEMSELVES FROM GETTING HIV AND TRYING TO MAINTAIN THEIR RELATIONSHIP. HONESTLY THERE IS INTERESTING RESEARCH ON COUPLES AND FAMILIES THAT LIVE WITH LONG TERM ILLNESS AND TRY TO UNDERSTAND THE STRUGGLE BETWEEN BALANCES DISEASE RISK AND TRYING TO MAINTAIN A HEALTHY RELATIONSHIP. WE SEE A LOT OF GENDER PATTERNS IN NEGOTIATE RISK IN SOUTH AFRICA. A LOT OF TRUST, A LOT OF TRUST, PEOPLE BELIEVE IF YOU TRUST YOUR PARTNER YOU WON'T GET HIV AND TRUST IS USUALLY THE WAY TO NEGOTIATE A HEALTHY RELATIONSHIP. WE ALSO SEE A DIFFERENT PATTERN IN NEGOTIATING RISK IN MEN AND WOMEN IN SOUTH AFRICA AND WE SEE THAT WOMEN EXPECT AND ACCEPT THAT MEN WILL HAVE MULTIPLE PARTNERS AND BOTH MEN AND WOMEN EXPECT WOMEN TO INITIATE DISCUSSIONS ON HIV AND RISK IN THEIR RELATIONSHIPS AND IF YOU HAVE AN EMPOWERED RELATIONSHIP THAT'S HARD FOR A WOMAN TO NEGOTIATE AND DISCUSS, INITIATE DISCUSSIONS ON HIV AND RISKING THEIR RELATIONSHIPS. SO THOSE MEN AND WOMEN ARE PEM eWOMEN ARE EXPECTED TO INITIATE DISCUSSIONS ON HIV. THERE'S AN APPROACH FROM THE (INDISCERNIBLE) STUDY WE'RE DOING PART OF THIS COMMUNITY RANDOMIZED STUDY WE'RE DOING AN ETHNOGRAPHIC COHORT, TRYING TO LOOK AT THIS COURSE IN FAMILIES OVER TIME TO SEE HOW THIS INTERVENTION NOT ONLY IMPACTS ON HIV INCIDENTS BUT HOW IT CHANGES COMMUNITY NORMS AND STANDARDS. THIS IS A QUOTE FROM SOMEONE IN (INDISCERNIBLE) THAT SAY IT'S NOT EASY FOR A MALE PERSON TO TEST BECAUSE COMMON MEN DO -- THE PROBLEM IS FEMALES SO THIS IS AN ISSUE THAT WOMEN ARE EXPECTED TO TEST AND THERE'S LITTLE PROSI TESTENING SOUTH AFRICA. IN TERMS OF TRYING TO DRIVE DOWN THE BURDEN OFTEN HIV IN SOUTH AFRICA, WE HAVE A NATIONAL STRATEGIC PLAN COMING TO -- THE CURRENT PLAN TO BE REVISED, THEY HAD A BOLD APPROACH IN THE ADMINISTRATION TO TRY TO REDUCE TRANSMISSION, TO REDUCE INFECTIONS TO MOTHER TO CHILD TRANSMISSION RATES, HIV INFECTION THROUGH BLD AND BLOOD PRODUCTS, TO INCREASE COVERAGE OF EZT AND ADHERENCE AND TO SUPPORT THE DEVELOPMENT OF MICROBICIDE AND AIDS VACCINE. IN TERMS OF MOTHER TO CHILD TRANSMISSION, YOU CAN SEE WE ARE HIGH PREVALENCE OF HIV AMONG WOMEN IN SOUTH AFRICA. THIS IS BY 30% OF ALL PREGNANT WOMEN IN SOUTH AFRICA HAVE HIV AND INTERVENTIONS TO PREVENT MOTHER TO CHILD TRANSMISSION HAVE TO ADDRESS THE ISSUE OF INEFFECTIVE WOMAN AND AS WELL AS PREVENTION. IN TERMS OF (INAUDIBLE) PROGRAM OVER THE LAST EIGHT YEARS WE MANAGED TO ALMOST A QUART OF A MILLION WOMAN WITH HIV TESTING AND HAVE SEEN OVER TIME ONCE INTRODUCED MORE COMPLICATED REGIMEN TRANSMISSION RATES ARE GOING DOWN. THIS IS ONE OF -- I HAVE THREE BEAUTIFUL SLIDES, THIS IS ONE THAT I'M GOING TO SHOW YOU. IN THE POPULATION WE'RE STARTING TO SEE REDUCTIONS IN TRANSMISSION RATES. YOU CAN SEE FROM OUR PROGRAMMING THAT INTRODUCING MORE COMPLICATED INDIVIDUALIZED INTERVENTIONS WE CAN GET TO RATES OF UNDER 2.8% BUT AS YOU CAN SEE THAT MEANS WE STILL HAVE WORK TO DO AN POSSIBLY THE ROLE OF NEUTRALIZING ANTIBODIES HELP TO ELIMINATE OR ERADICATE PEDIATRIC HIV. SO IT'S -- THIS IS MY SECOND MOST AMAZING SLIDE. THIS IS TO HAVE A LOOK AT A POPULATION TO SEE WHAT'S HAPPENING TO INFANT MORTALITY RATE IN SOUTH AFRICA. YOU CAN SEE THAT FROM 1976 TO 1990, THERE WAS A REDUCTION IN INFANT MORTALITY RATE IN SOUTH AFRICA. AROUND '92 TO '39 WE SAW REVERSAL IN TRENDS AND SAW INFANT MORTALITY RATES EQUIVALENT TO 1984 AND AROUND 2000. AND SLOWLY ROLLING OUT OF MOTHER TO CHILD TRANSMISSION WE'RE STARTING TO SEE A REDUCTION IN INFANT MORTALITY RATES. THIS IS THE SECOND POPULATION BASED DATA THAT SHOWS THAT INCREASING COVERAGE ADDRESSING HIV TESTING IN PREGNANCY DELIVERING INTERVENTIONS CAN ACTUALLY HAVE A IMPACT TO THE POPULATION LEVEL ON CHILD SURVIVAL. WE STILL HAVE A LOT OF WORK TO DO IN TERMS OF HIV AND MORBIDITY AND MORTALITY IN SOUTH AFRICA. THIS IS DATA FROM OUR STUDY SHOWING THAT A LOT OF WOMEN DO CHOOSE TO FORMULA FEED IN HIV INFECTED INFANTS FORMULA FED IS DETRIMENTAL TO YOUR -- YOU HAVE ADVERSE EVENTS, YOU INCREASE YOUR MORE BITTY AND WE NEED INTERVENTIONS THAT COULD PREVENT BABIES BEING FORMULA FED. BASED ON THIS DATA WE STARTED A STUDY LOOKING AT RELACTATION. SO IF WOMEN DON'T BREAST FEED BABIES AT FOUR WEEKS OF AGE WHEN WE DIAGNOSE HIV INFECTION IN THE CHILDREN WE INITIATE RELACTATION AND SEE IF WE CAN GET THE BABIES TO START, AND THE DATA IS -- WE PRESENTED THE DATA IS LOOKING VERY GOOD AND VERY PROMISING. IN TERMS OF MORTALITY IN THE 1st 100 DAYS OF LIFE, BEING HIV INFECTED INCREASES THE ODDS OF DYING BY FOUR FOAL IF YOU'RE CHILD AND MATERNAL LYREMIA. IN TERMS OF FORMULA FEEDING DID NOT IMPACT ON THE MORTALITY IN THIS STUDY. WE WERE FORTUNATE TO GET FUNDING. WE WERE (INDISCERNIBLE) ONE OF OUR STUDIES THAT WE DESIGNED AS PART OF THE PROGRAM WAS A STUDY TO LOOK AT OPTIMIZING (INDISCERNIBLE) CHILDREN, IN THE FIRST PART OF THE STUDY CHILDREN WERE RANDOMIZED TO RECEIVE EARLY TREATMENT AT BETWEEN 4 AND 12 WEEKS OF AGE OR TO RECEIVE THE SENATE GUIDELINES AVAILABLE AT THE TIME. YOU CAN SEE THAT EARLY TREATMENT IRRESPECTIVE OF CD4 COUNT AND VIRAL LOAD REDUCE MORTALITY BY TWO-THIRDS AND HAS CHANGED NATIONAL AND INTERNATIONAL GUIDELINE AND MOST CHILDREN NOW SOUGHT EARLY TREATMENT BASED ON THE STUDY. SO I E SHOW YOU GOOD DATA IT DOES WORK AND EARLY INTERVENTIONS CAN IMPROVE SURVIVAL. I'M NOT GOING TO TALK ABOUT THE ROLE OF ANTIRETROVIRAL THERAPY -- I WILL NOW TALK ABOUT ANTIRETROVIRAL THERAPY AND IMPACT ON DEATH IN SOUTH AFRICA. I TOLD YOU BEFORE WIFE A HUGE PROGRAM AND IT IS LARGEST TEACHING -- WE HAVE A HUGE PROGRAM BUT WE STILL LAG BEHIND, AS MENTIONED BEFORE ONLY A THIRD OF PEOPLE WHO NEED ANTIRETROVIRAL THERAPY ON ARE IT. WHEN YOU LOOK AT INTERVENTIONS LIKE LOOKING AT SOME OF THE ISSUES AROUND RUNNING A TREATMENT PROGRAM IN SOUTH AFRICA, WE KNOW THAT TREATMENT STARTED TO BE SCALED UP IN ABOUT 2000. AND THIS WAS A POLITICAL DECISION, WE WERE GOING TO A NEW ELECTION AND THERE WAS A LOBBY THAT FORCED THE GOVERNMENT TO START DELIVERING ANTIRETROVIRAL THERAPY AND THAT STARTED SLOWLY IN ABOUT 2000 AND ONLY RECENTLY HAVE WE MANAGED TO SCALE UP TO 1.2 MILLION. THERE HAVE BEEN A LOT OF BENEFITS OF NOT PROVIDING THE THERAPY IN A RAPID FASHION. PEOPLE HAVE ESTIMATED THE LOST BENEFITS. IT'S ESTIMATED THAT MORE THAN 330 LIVES OR APPROXIMATELY 2.2 MILLION PERSONAL LIVES WERE LOST. 35,000 BABIES BORN WITH HIV WHICH RESULTS IN 1.6 PERSON YEARS LOST BY NOT IMPLEMENTING A REGIMEN AND THE TOTAL BENEFITS ARE AT LEAST 3.8 MILLION PERSON YEARS FOR THE PERIOD OF 2000 AND 2005. S THAT GREAT SADNESS FOR A COUNTRY. AND IMPORTANT TO ADDRESS ISSUES OF ALV ROLL-OUT. LOOKING AT REASONS FOR NOT TAKING NOW THAT WE DO HAVE ANTIRETROVIRAL THERAPY AVAILABLE, WE LOOK AT DATA FROM MY -- WE OFFER ABOUT 45 PEOPLE RECEIVE VCT AT OUR SITE AND WE DO CD4 COUNTS, WE LOOK AT PEOPLE ANTIRETROVIRAL TREATMENT TO PEOPLE REQUIRE IT. AND LOOKING AT OUR DATA WE FOUND ONE IN FIVE LEGIBLE TREATMENT, ACTUALLY REFUSE TREATMENT. THE REASON THEY REFUSE IS THEY STILL FEEL HEALTHY. THEY AREN'T ABLE TO DISCLOSE, THEY WORRY ABOUT THE SIDE EFFECTS, ABOUT BEING ABLE TO HIT THE REGIMEN, THEY FOUGHT CULTURAL BLIEF BELIEFS AND STIGMA SO EVERY TRIAL AN ROLL-OUT TREATMENT IN SOUTH AFRICA, SOME OF THE BEHAVIORAL ISSUES WE HAVEN'T ADDRESSED, REASONS PEOPLE TAKE DRUGS, NEED TO BE PAY MORE ATTENTION TO THIS. BUT THE GOOD THING ABOUT ANTIRETROVIRAL THERAPY IN SOUTH AFRICA THOUGH IT WAS DELAYED A DECLINE IN DEATHS IN SOUTH AFRICA, THE FIRST REAL EVIDENCE THAT'S IN FACT THE PROGRAM IS STARTING TO SLOWLY IMPACT ON UNREPORTED DEATHS IN SOUTH AFRICA. SO ABOUT TREATING IN THIS CONTEXT? TEST AND TREAT HAS BEEN -- SCIENTISTS BELIEVE TEST AND TREAT COULD IMPACT ON PREVENTION AND TESTING WHOLE POPULATIONS FOR HIV AND TREATING THOSE INFECTED MAY SLOW THE SPREAD OF HIV. THIS IS AN IMPORTANT APPROACH, TO ADDRESS AND THE CHALLENGES OF PROVIDING A TEST AND TREAT PROGRAM IN A HIGH PREVALENCE COUNTRY NEEDS TO BE TESTED. SO THERE ARE STUDY, FEASIBILITY STUDIES, ACCEPTABILITY STUDYING BEING PLANNED TO SEE WHAT ONE DOES IN A HIGH PREVALENT AREA IN A POOR COUNTRY WHAT RESOURCES DO YOU NEED. AND THERE ARE SOME CHALLENGES THAT NEED TO BE TESTED. ONE TO SEE WHAT THE FEASIBILITY AND ACCESSIBILITY IS. AND WHAT THE COST IS, CAN GOVERNMENTS AFFORD THIS, WE KNOW MOST AFRICAN GOVERNMENTS HAVE -- CAN ONLY AFFORD SICK YEARS DOLLARS PER PERSON PER YEAR FOR HEALTH. WE NEED THE ADDRESS ISSUES OF EFFICACY, SUSTAINABILITY AND ISSUE OF TOXICITY. THIS IS ANOTHER REASON NOT TO DO IT BUT THESE ARE THINGS WE TEST WHEN LOOKING AT THE FEASIBILITY STUDIES. SO WHAT ABOUT REDUCING THE INFECTION AND WHAT ABOUT SUPPORTING THE DEVELOPMENT OF MICROBICIDES AND AIDS VACCINES IN SOUTH AFRICA? THE GERMAN (INAUDIBLE) AND EVERY TIME THERE'S A NEW PREVENTION TRIAL THAT COMES IN, WE EDIT AND LOOK AT IT. WE HAVE FOUR INTERVENTIONS THAT HAVE SHOWN TO ROBUST EFFICACY. THE HIV VACCINE IN THAILAND SHOW 31% EFFICACY, WE HAVE TWO ANTIRETROVIRAL REGIMEN, OTHER SYSTEMIC OR TOPICAL ANTIRETROVIRALS THAT ARE SHOWN TREATMENT EFFECTS AND WE ALSO HAVE CIRCUMCISION SHOWN TO REDUCE TRANSMISSION IN MEN SUBSTANTIALLY. AND ANY INTERVENTIONS IN SOUTH AFRICA NEED TO LOOK AT THIS AND NEED TO APPLY THEM AND SEE HOW THEY WORK. AS FAR AS CIRCUMCISION IS CONCERNED, THE GOVERNMENT HAS STARTED TO ROLL OUT MALE CIRCUMCISION. WE WITH FUNNING FROM PEPFAR HAVE A CIRCUMCISION CLINIC IN A HOSPITAL WHICH IS OUR MINISTER OF HEALTH WHO CAME TO OPEN UP ON THE SAME DAY HE DID TEN CIRCUM CIRCUMCISIONS HIMSELF, SINCE STARTING WE GOT TO 220 PER WEEK. ON FRIDAY WE HAD ALREADY DONE HALF THE MONTH AND WE HAVE ALREADY DONE SO MUCH. THE BIG CHALLENGE AROUND CIRCUMCISION IS HOW TO ROLL IT OUT IN A -- WE KNOW ONE HIV INFECTION CAN BE AVERTED FOR EVERY (INDISCERNIBLE) THAT YOU DO EVERY DAY. WE HAVE 10 MILLION MEN IN SOUTH AFRICA THAT NEED TO BE CIRCUMCISED AND WE HAVE A COHORT OF BOYS OF HALF A MILLION. SO THE CHALLENGE OF TRYING TO ROLL OUT MALE CIRCUMCISION IS MORE OVERWHELMING THAN ROLLING OUT ANN RETROVIRAL THERAPY BUT ARE VERY IMPORTANT INTERVENTION THAT HAS TO HAPPEN IF WE WANT TO MAKE SUSTAINED ROADS INTO PREVENTING INFECTIONS IN SOUTH AFRICA. AS FAR AS THE HIV VACCINE MICROBICIDE RESEARCH IS CONCERNED THE ONLY WAY THE TANOFOVIR GEL MICROBICIDE STUDY DATA THAT WAS PRESENTED LAST YEAR, THIS IS A CULTURAL DEPENDENT MOI CROAB SIDE THAT'S ADDED BEFORE AND AFTER SEX AND NOT MORE THAN WITHIN TWO DAYS IN 24 HOURS. HERE YOU SEE THE EFFICACY OF 39% AT 15 MONTHS FOLLOW-UP AND YOU SEE GOOD EFFECTS OF 50%. YOU SEE A DECLINE IN THE DURABILITY OF THE INTERVENTION. AND WE HAVE THE ADDRESS ISSUES OF ADHERENCE AND ISSUES OF WHY WE ARE UNABLE TO SUSTAIN CONSISTENT FUTURE STUDIES HAVE TO ADDRESS WAYS OF INTERVENING AND I THINK THIS DATA ALSO SHOWS THE IMPORTANCE OF HAVING A VACCINE WHO GIVE IT A FEW TIMES, WHO DIDN'T HAVE TO WORRY ABOUT ADHERENCE IN THE LONG TERM. AS MENTIONED BRIEFLY ADHERENCE ISSUE, YOU CAN SEE THAT THE EFFICACY DOES DECLINE IN OVER PEOPLE BE ADHERENT TO NOT, VERY IMPORTANT WHEN WE DESIGN VACCINE TRIALS TO TRY AND GAUGE THE EFFECTS OF HEROES AND VACCINE EFFICACY. IN TERMS OF TA NOVEMBER VEER GEL STUDIES, WHEN WE HAVE THE STUDY THAT WE HOPE WILL STOP ENROLLMENT, COMPLETE ENROLLMENT MAY THIS YEAR, WE EXPECT RESULTS TO BE AVAILABLE END OF NEXT YEAR OR 2013. THERE'S AN EFFECT STUDY PLANNED IN JULY THIS YEAR IN SOUTH AFRICA IN SOUTH AFRICA AND THIS STUDY HOPE TO BE FULLY ENROLLED BY FEBRUARY OF NEXT YEAR AND HAVE ENOUGH IN TERMS OF FUTURE DEVELOPMENT, THE NEXT ROUND OF MICROBICIDE GELS WILL BE ARRANGED AND WILL HAVE AN ANTIBODY AND A RING THAT ONE CAN TEST IN MICROBICIDES. IN TERMS OF HIV VACCINE RESEARCH I'M GOING TO MENTION THREE TRIALS, SOUTH AFRICA HEAD WAS INVOLVED IN SIX PHASE 1, PHASE 2 STUDY AND ONE PHASE 2B STUDY. I'M GOING TO MENTION DATA ON THE VRCH BETWEEN 204 STUDY, THIS WAS CONDUCTED IN SOUTH AFRICA AS WELL AS OTHER SITES. THE PHASE 2B STUDY AND VACCINE STUDY THAT JUST COMPLETED ITS FIRST PHASE OF VACCINE REGIMEN. SO THE STUDY YOU CAN SEE THAT JUST HALF OF PARTICIPANTS CAME FROM SOUTH AFRICA, A LOT OF THESE PARTICIPANTS WERE FEMALE, AND YOU CAN SEE IN TERMS OF PRE-EXISTING IMMUNITY PARTICULARLY IN SOUTH AFRICA, MOST PARTICIPANTS HAD PRE-EXISTING AND THIS SHOWS YOU THE REST OF THE DEMOGRAPHICS AT THE END OF THE STUDY AROUND SEVERAL PEOPLE AROUND HIV. THIS IS A SLIDE, I WANT TO GIVE YOU THE IMPRESSION OF THE IMMUNE RESPONSES THAT WE HAVE SEEN SO THE TOP SHOWS THE CD8 AND CD4 RESPONSES FOR ALL PARTICIPANTS AND THE DATA ON YOUR RIGHT LOOKS AT THE RESPONSE ACCORDING TO (INDISCERNIBLE) STATUS. YOU CAN SEE THEY ARE GOOD RESPONSES TO THIS VACCINE AND MOST (INDISCERNIBLE) THERE'S A SLIGHT RESPONSE IN PEOPLE WITH PREEXISTING IMMUNITY TO ADINO5. GOOD POLYFUNCTIONALTY. I SEE THE CYTOKINE RESPONSES 1 AND 2 CYTOKINES, SURE YOU HAVE SEEN THE DATA BEFORE, SO I WOULDN'T DWELL TOO MUCH ON IT. IN TERMS OF ANTIBODIES TO TR-1 VIRUSES WE SEE RESPONSES, TO MN AND SF-162. NO RESPONSES TO VIRUSES. IN TERMS OF THE STUDY WE ADDED THE FIRST PHASE 2B TEST OF CONCEPT TRIAL. THIS WAS A SUBTYPE B IN A SUBTYPE C AREA, WE WERE LERSED IN LOOKING AT AREAS OF PROTECTION. THE THOUGHT IN THE STUDY WAS A HUGE THING, IT WAS BIG NEWS, WE HAD TO STOP WHEN THE RESULTS OF THE STUDY CAME OUT AND SPEND TIME UNBLINDING AND FOLLOWING THEM UP. IN TERMS OF DATA FROM THE PAMBILI STUDY YOU CAN SEE THE VACCINE DID NOT PREVENT HIV INFECTION. WE HAVE HIGH INCIDENCES, 7% IN THE VACCINE ARM VERSUS THE PLACEBO ARM AND MEN 3% IN VACCINE ARM VERSUS 2% IN PLACEBO ARM. SO LOOKING AT VIRAL LOAD SET POINTS OVERALL THERE WAS NO DIFFERENCE IN VIRAL LOAD SET POINT BETWEEN RECIPIENTS. WE NEED MORE DATES ON GENDER BUT THERE WAS A NON-STATISTICALLY DIFFERENT LOWER VIRAL LOAD SET POINT IN WOMEN WHO -- WOMEN WHO BECAME HIV INFECTED AND RECEIVED THE VACCINE. INTERESTING, INTERESTING DATA IN CD4 DECLINE, WHEN YOU LOOK AT A WOMAN WHO BECAME HIV INFECTED AND WHO RECEIVED THE VACCINE EARLY ON IN THE FIRST THREE MONTHS POST INFECTION, THERE SEEMS TO BE AN EFFECT LESS WOMEN IN THE VACCINE ARM AS COMPARED TO PLACEBO, FOR THE SPECIFICITY OF LESS THAN 350. TRYING TO UNDERSTAND JN DER EFFECTS AS FAR AS VACCINE EFFECTS IN PHASE 2B STUDIES. THIS IS VACCINES, THIS IS SPONSORED BY THE NIH AND THE AFRICAN GOVERNMENT WE HAVE THE BACKBONE IN OUR DNA AND WE JUST COMPLETE THAT IN THE PHASE 1 STUDY SHOWS GOOD RESPONSE, OUR RESPONSE IS ON TO SHEBE. YOU CAN SEE THE DATA AFTER THE FIRST NCA BOOST, MOST RESPONSES IS OFF AS FAR AS CD4 IS CONCERNED. CD8, AGAIN, NOT TOO BAD RESPONSES AND MOST OF THE RESPONSES ARE ONC OR GAG. FAIRLY GOOD POLYFUNCTION, JUST IN TERMS OF LOOKING AT THE RESPONSES JUST IN SUMMARY, YOU CAN SEE THAT THE RESPONSES ARE NOT BAD THE NORTH AMERICAN AND SOUTH AFRICAN PARTICIPANTS SEEM TO BE SOME SUBDUED RESPONSES WE NEED TO INVESTIGATE FURTHER. IN TERMS OF VACCINE DESIGN, I HAVE SHOWED YOU A LITTLE BIT, ALL OF YOU ARE AWARE OF THE FIELD, WE NEED TO BE MORE AGGRESSIVE IN VACCINE TRIALS. WE NEED TO LOOK AT MULTIPLE VACCINES AND RAPIDLY SEE WHAT WORKS AND DROP THE ONES THAT DON'T WORK. WE NEED TO CONCURRENTLY MAKE SURE THAT WE ARE DOING THE -- NOT WAIT FOR THE CORRELATES WORK FOR AFTER THE STUDIES, THEY HAVE TO DO IT IN A CONCURRENT FASHION. WE HAVE MADE SOME HVTN THAT ONE NEEDS TO ONE NEEDS TO RUN COMPARATIVE ARM TO INCREASE CORRELATE AND PROVIDE RANK AND ORDER FOR VACCINES THAT NEED TO BE PRIORITIZED FOR LICENSURE AND FOR THE DEVELOP. AND HOPEFULLY THIS APPROACH GOING FORWARD WILL REVOLUTIONIZE THE VACCINE FIELD AND HELP US GET VACCINES THAT ARE EFFICACIOUS. SO IN TERMS OF OUR POX PROTEIN DEVELOPMENT PLAN, WE HAVE A RESEARCH PLAN AND LICENSE PLAN AND THIS TIME LINES TO DEVELOP BOTH THE LICENSURE AND THE DEVELOPMENT STUDY IN SOUTH AFRICA AND HOPEFULLY THIS WILL START TOWARD THE END OF 2012 AND 2013. IN SUMMARY, I HAVE TWO MORE SLIDES LEFT. WE KNOW FROM THE DATA AND FROM THE DISCUSSIONS WE HAD AT THE VRC THIS MORNING, THAT THE LANDSCAPE FOR FUTURE OF HIV PREVENTION TRIALS IS CHANGING, AND CHANGING RAPIDLY AND SOON WE KNOW WE WILL HAVE LOTS OF PREVENTION INTERVENTIONS TO EMPLOYING THE STUDIES WHICH WILL LOWER THE HIV INCIDENCE. WE HAVE TO DESIGN STUDIES THAT INCORPORATE THE NEW INTERVENTIONS. AND WE WILL HAVE TO LOOK HOW WE COMPARE WHAT THE BACKBONE, THE EQUIVALENT STUDIES THE SUPERIORITY, WHAT WILL BE THE IMPACT ON HIV PATHOGENESIS AND PARTICULARLY OUR VACCINE TRIAL INPUT AT THE SET POINT AND NATURAL HISTORY. AND ALSO MOST IMPORTANTLY, AND PARTICULARLY FOR THE NIH, THESE INTERVENTIONS THOUGHT TO BE PREVENTABLE WILL IMPACT ON THE COST OF TRIALS AN COMPLEXITY OF TRIALS THAT NEED TO BE ROLLED OUT IN THESE SETTINGS. IN CONCLUSION WE BEAR THE BURDEN OF HIV AND CAN CONTRIBUTE TO ITS SOLUTIONS. WE HAVE ALREADY SEEN THAT PMTC IS A SUCCESS IN SOUTH AFRICA. THAT ALTHOUGH (INDISCERNIBLE) EXTRAORDINARY SOURCES OR WE'RE BEGINNING TO SEE THE DECLINE OF DEATHS IN SOUTH AFRICA. WE NEED TO RAPIDLY ROLL OUT MALE CIRCUMCISION AND LOOK AT NEONATAL CIRCUMCISION AS AN OPTION TO PREVENT HIV PREVENTION IN A 20 YEAR PERIOD. WE DO EXPECT THE GEL TO BE LASTING AROUND 2014 TO 2017 AND WE DO HOPE FOR VACCINE EFFICACY TRIALS TO START IN 2012, LATE 2012, EARLY 2013. I WOULD LIKE TO ACKNOWLEDGE THE HVTN, LARRY AND JIM AND JULIE AND MARY ELLEN FOR THEIR SUPPORT, THE AIDS VACCINE INITIATIVE, WILLIAMSON AND CAROL LYNN WILLIAMSON WHO DEVELOPED A VACCINE, (INDISCERNIBLE) FROM MY UNIT, NEAL MARTINSON, (INDISCERNIBLE) WHO DID THE SHARE STUDY AND THE HVTN STUDY TEAM AND THE CO-CHAIRS, GAVIN CHURCHYARD AND MIKE KEEFER. THANK VERY MUCH. [APPLAUSE] >> IT'S MY GREAT PLEA THUR STHOWR INTRODUCE THE LAST SPEAKER TODAY, DAVID BALTIMORE. IN THE INTEREST OF TIME I WON'T GO THROUGH ALL HIS CAREER EXCEPT OBVIOUSLY MENTION THAT HE WAS CO-RECIPIENT OF THE NOBEL PRIZE FOR THE DISCOVERY OF REVERSE TRANSCRIPTASE. HE IS HEADED THE WHITEHEAD INSTITUTE. THE ROCKEFELLER UNIVERSITY IN CAL TECH. HE'S CURRENTLY A PROFESSOR OF BIOLOGY AT CAL TECH WHERE HE NOW RUNS THE LAB, THAT IS INVESTIGATING WAYS TO GENETICALLY AUGMENT IMMUNE RESPONSES TO HUMAN PATHOGENERALS. -- PATHOGENS. HE'S PUBLISHED OVER 600 PAPERS, HE HAS INFLUENCED SCIENTIFIC THOUGHT AND SCIENTIFIC POLICY BOTH WITHIN THE UNITED STATES AND AROUND THE WORLD. AND I THINK THAT WITHIN THIS LARGE PRODUCTIVE CAREER, IF YOU WERE TO ASK HIM WHAT WAS THE MOST REWARDING PART OF HIS CAREER I'M SURE HE WOULD SAY IT WAS THE YEARS SPENT TRAINING GARY NABEL IN HIS LAB. [LAUGHTER] >> SO WE ARE VERY PLEASED THAT HE HAS FOR THE LAST TEN YEARS BEEN PART OF HEADING UP OUR SCIENTIFIC ADVISORY WORKING GROUP SO THAT WE CAN LEARN FROM HIS VAST EXPERIENCE. WE'RE ALSO VERY GRATEFUL THAT HE'S AGREED TO COME HERE AND ANCHOR OUR TEN YEAR SYMPOSIUM. DAVID. [APPLAUSE] >> IT IS CERTAINLY TRUE THAT THE MOST SATISFYING PART OF MY CAREER IS TRAINING NOT ONLY GARY NABEL, A NUMBER OF OTHER PEOPLE, SOME OF WHOM ARE HERE. I TAKE SERIOUSLY THE FACT THAT I HAD TO ANCHOR THIS SYMPOSIUM AND SO I PRODUCED SOMEWHAT PRETENTIOUS TITLE, SET ON A BEAUTIFUL PHOTOGRAPH OF THE SAN ANDREIAS FAULT OVERLOOKING LOS ANGELES. THERE'S NO SIGNIFICANCE TO THE SAN ANDREIAS FAULT IN THIS PARTICULAR SETTING. BUT I THOUGHT I WOULD SUM UP A LITTLE BIT PARTLY AS A BACKGROUND TO WHAT I WANT TO TALK ABOUT. THE SEARCH FOR AIDS VACCINE GOES BACK TO THE MID 1980s. IT'S NOW MORE THAN 30 YEARS, IT'S EASY IN EARLY DAYS TO STATE THE PROBLEM. HOW DO YOU ELICIT FROM THE HUMAN ANTIBODY GENE REPERTOIRE POPULATION OF ANTIBODIES THAT CAN NEUTRALIZE HIV IN ALL OF ITS VARIETY? THAT WAS A QUESTION BASICALLY MODELED ON HOW VACCINES TO OTHER VIRUSES WERE THOUGHT TO WORK AND HOW THEY WERE DEVELOPED. IT SLOWLY THEN BECAME EVIDENT THAT HIV WAS WAY AHEAD OF US. IT WAS VERY HARD TO NEUTRALIZE THIS VIRUS AND THAT IT WAS EXTREMELY VARIABLE. ONE PHASE 3 CLINICAL TRIAL WAS RUN TO LOOK AT THE ABILITY OF ANTIBODY TO PROTECT AND IT FAILED. SO TIME TO CHANGE THE QUESTION. SO WE ASKED WHAT WAS REALLY AN UNPRECEDENTED QUESTION SINCE NO ONE EVER REALLY SERIOUSLY ENTERTAINED IT COULD WE ELICIT PROTECTIVE T-CELLS USING THE GENES IN THE T-CELL REPERTOIRE, SO I WOULD CLASS THIS AS A HAIL MARY AND IT WILL ALSO SEEM TO FAIL IN THE ONE PHASE 3 CLINICAL TRIAL. THEN WE COMBINE THESE FAILED CONCEPTS IN ANOTHER TRIAL AND WE GOT A SIGNAL INDICATING PROTECTION IN THE TRIAL. PROBABLY TRANSIENT BUT ENOUGH TO GIVE A FRESONG OF EXCITEMENT TO THE FIELD, A QUIVERING SINCE THAT HAS ENERGIZEED INVESTIGATIONS WORLDWIDE. I THINK IT HAS TO BE SEEN THIS EXPERIENCE IS HUMBLING. I PERMLY CONSIDERED THE OUTSTANDING FAILURE OF MODERN BIOLOGY TO DELIVER TO SOCIETY A VACCINE AGAINST HIV WHEN THE NEEDS OF SOCIETY AS WE JUST HEARD IN GREAT DETAIL IS SO GREAT. BUT WE HAVE TO REMEMBER THAT IT WAS NOT A FAILURE OF WILL NOR SKILL, IT WAS A FAILURE CAUSED BY THE INVENTIVENESS OF NATURE BEING AHEAD OF OUR TECHNOLOGY. THE WONDERFUL THING IS AS PARTICULARLY EMBODIED IN THE VRC, THAT OUR COMMUNITY HAS NOT GIVEN UP. BUT RATHER AT EACH TIME OF FAILURE OR UNCERTAINTY HAS REDOUBLED ITS EFFORTS LETTING A THOUSAND FLOWERS BLOOM, LETTING NEW IDEAS COME TO THE FLOOR, NEW DIRECTIONS APPEAR. AND OFTEN RESUSCITATING OLD IDEAS WITH NEW TECHNOLOGY, PARTICULARLY GLAD TO HAVE SEEN ASHLEY HAASE TALKING ABOUT RON DEROSIER'S QUITE DRAMATIC RESULTS OF MANY YEARS AGO THAT WE HAVE NOT UP TO THIS TIME BEEN ABLE TO UNDERSTAND. I THINK THE MOST IMPORTANT RECENT ATTITUDENAL CHANGE HAS BEEN A REFOCUS ON ANTIBODIES. BASICALLY LEARNING THE LESSON FROM THE PAST WHICH I THINK IS THE CORRECT LESSON THAT ANTIBODY IS ABLE TO GIVE US PROTECTION AGAINST VIRUS THE WAY NOTHING ELSE CAN. THE FACT REMAINS IS THERE IS NO AFFECTIVE STRATEGY FOR ELICITING ANTIBODIES WITH BROAD NEUTRALIZING ACTIVITY AGAINST HIV FROM THE REPERTOIRE OF GENES THAT WE INHERIT. HOWEVER, THERE ARE BROADLY NEUTRALIZING ANTIBODIES, GARY GAVE US SOME VERY GOOD EXAMPLES OF THOSE AND THERE ARE MANY MORE THAT HAVE IT'SLATED AND BEING ISOLATED, EACH ONE BROADER, MORE EFFICACIOUS THAN THE NEXT AND THEY'RE BEING STUDIED IN REMARKABLE DETAIL. SO THEY ARE REALLY FABULOUS POTENTIAL SOURCES OF PROTECTION AGAINST HIV INFECTION. , SO THE ISSUE BECOMES HOW DO WE BUILD ON THIS DEMONSTRATION THIS THE HUMAN REPERTOIRE CAN MAKE POTENTIALLY PROPHYLACTIC ANTIBODY. MAYBE WE CAN DESIGN AN IMMUNOGEN THAT WILL ELICIT THESE WONDERFUL ANTIBODIES. AN UNPRECEDENTED APPROACH, NO ONE HAD EVER DONE THIS DESIGNED AN IMMUNOGEN TO GIVE RISE TO A SPECIFIC CLASS OF ANTIBODY. AND AGAIN, GARY AND OTHERS HAVE GIVEN US A PICTURE HOW WE MIGHT TRY TO DO THAT. AT LEAST THE BROADLY NEUTRALIZING ANTIBODIES ARE ALL HIGHLY MUTATED. I DON'T KNOW THE DATA ON MORE RECENT ONES THAT HAVE BEEN ISOLATED. THUS YOU HAVE TO IMAGINE WHAT IS A DESIGNED IMMUNOGEN HAS TO DO IS NOT TO REPRODUCE A PROCESS BUT REPRODUCE A PATH BECAUSE THE EVOLUTION OF HIGHLY MUTATED ANTIBODY IS A SERIAL SELECTION PROCESS. AND TO GET A PATH TO A VERY RARE PRODUCT INVOLVES CARE CARRYING THANK THROUGH A PO PROCESS OF MUTATION AND SELECTION. SO IT IS A TRULY DIFFICULT PROCESS. SO I THINK WE HAVE TO SAY, WE HAVE TERRIFIC ANTIBODIES, AND MAYBE THERE ARE OTHER WAYS OF USING THEM. WE HAVE SEEN THE MOST OBVIOUS ANSWER TO THAT QUESTION WHICH IS WE'LL INJECT THEM. AND TO DO THAT WE HAVE TO MANUFACTURE THEM, BUT WE CAN DO THAT AND THERE'S A WHOLE INDUSTRY OF PEOPLE WHO KNOW HOW TO MANUFACTURE MONOCLONAL ANTIBODIES. IT WILL PROBABLY BE EXPENSIVE TO GET ENOUGH ANTIBODY, THERE ARE COMPLIANCE ISSUES WITH ANYTHING THAT HAS TO BE DONE OVER AND OVER, TAKEN OVER AND OVER AGAIN. SO IT MAY NOT BE A PERFECT SOLUTION BUT IT CERTAINLY IS A SOLUTION THAT OUGHT TO BE AND IS BEING FOLLOWED. BUT IF WE CAN'T FIGURE OUT HOW TO ELICIT THESE, IF WE SAY INJECTION IS TOO DIFFICULT A ROUTE TO GO ON, ARE THERE OTHER WAYS THEY CAN BE USED. AND ONE WAY WOULD BE TO DIRECT THE BODY TO MAKE THESE MONOCLONAL ANTIBODIES. AFTERALL, WE HAVE THE GENES FOR THEM, WE KNOW HOW TO PU GENES IN TO CELLS TO GET GENES TO MAKE PROTEINS, SO IT'S A STRAIGHT FORWARD SUGGESTION. IT COULD BE A FORM OF FREE EXPOSURE PROPHYLAXIS BUT USING NOT DRUGS BUT GENES THAT ENCODE ANTIBODIES AS A DELIVERABLE. WE HAVE THINKING ABOUT THIS ISSUE FOR TEN YEARS. LONG BEFORE THESE ANN BODIES EXISTED SO WE WERE BEING OPTIMISTIC THAT THE TIME WOULD COME WHEN THERE WOULD BE ANTIBODIES AROUND AND IT COULD BE USED. WESK WORKING ON VARIOUS APPROACHES TO DO THIS. WE CALL THE OVERALL PROGRAM OF GENE MODIFICATION THAT WE'RE DOING FOR THE IMMUNE SYSTEM, ENGINEERING IMMUNITY AND IS RELEVANT TO CANCER AS WELL AS TO AIDS BUT I'LL JUST TALK ABOUT THIS GENE THERAPY FORM OF TRYING TO PROVIDE INFECTION AGAINST HIV. SO WE NEED A METHOD TO GENERATE THE CONTINUE WALL PRODUCTION OF DEFINED MONOCLONAL ANN BODIES IN THE BODY OVER PERIODS OF YEARS FOLLOWING A SINGLE GENE INJECTION. PREFERABLY FOR A LIFETIME. OUR ORIGINAL APPROACH WAS TO DO THIS BY PROGRAMMING BONE MARROW STILL CELLS WITH ANTIBODY GENES USING LENTIVIRUSES AS GENE CARRIERS. I STILL THINK THIS WAS THE BEST WAY TO GO BECAUSE IT WOULD INSERT THE GENE INTO THE NATURAL PROCESS OF DEVELOPMENT OF B CELLS AN ALLOW B CELLS TO MAKE ANTIBODIES, BUT NOW WE'RE TELLING THE B CELL WHAT TO MAKE RATHER THAN TRYING TO COAX IT TO MAKE THE THING WE WOULD LIKE TO HAVE IT MAKE. WE HAVE FOUND IN TRYING TO DO THIS THERE ARE LOGISTICAL PROBLEMS AND PROBLEMS I WON'T GO INTO WHAT THE PROBLEMS ARE. BUT I MUST SAY THE NO SOLUTION TO THIS PROBLEM HAS APPEARED IN MY LABORATORY AND I DON'T KNOW OF ANYBODY ELSE, I DON'T KNOW MANY THAT EVEN TRIED. SO WE HAVE TAKEN A MUCH MORE SLEDGE HAMMER APPROACH WHICH IS TO USE ADINOASSOCIATED VIRUS. AND I CALL THE SLEDGE HAMMER APPROACH BECAUSE WE GOT TO PUT A GENE INTO THE VIRUS AND THEN JUST SIMPLY PUT THE VIRUS INTO MUSCLE CELLS AND LET MUSCLE CELLS MAKE ANTIBODIES. SO ALL THIS WORK HAS BEEN EITHER DONE OR DIRECTED BY ALEX BALLAS, A VERY INVENTIVE POST-DOCTORAL FELLOW WORKING WITH ME. AS YOU CHOSE ADINO-ASSOCIATED VIRUS, FOR A NUMBER OF REASON, MOST OBVIOUS TO THE GENERAL FIELD AND THE REASON THAT ADINOASSOCIATED VIRUSES BEING USED BROADLY IN GENE THERAPY AND IN VARIOUS SETTINGS. IT'S NON-PATHOGENIC TO HUMAN, NON-INTEGRATING BUT IT SETS UP A PLASMID IN A CELL THAT THEN GIVES VERY LONG-LIVED EXPRESSION AFTER A SINGLE ADMINISTRATION OF VIRUS. IT GIVES EXCELLENT EXPRESSION OF PROTEIN, MESSENGER RNA AND PROTEIN, AND REMARKABLY MUSCLE CELLS ARE HAPPY TO SECRETE ANTIBODIES IF THEY'RE MADE INSIDE MUSCLE CELLS. I FIND THAT REMARKABLE. THEY HAVE ONE CLEAR NEGATIVE CHARACTERISTIC, THEY'RE VERY SMALL. THEY HAVE A CARRYING CAPACITY OF 5 KB. ENOUGH TO PUT AN ANTIBODY GENE IN BUT NOT MUCH MORE. SO WE IN THINKING ABOUT THIS DECIDED TO GO TO THE EXPERT, WHO IS JIM WILSON, AT UNIVERSITY OF PENNSYLVANIA AND WE WENT TO HIS LAB AND LEARNED FROM HIM HOW HE MANIPULATES AAV AND THAT INVOLVES A PACKAGING SYSTEM THAT ALLOWS THE VIRUS TO BE PACKAGED USING BOTH ADINOVIRUS HELPER GENES WHICH ARE NECESSARY FOR THE REPLICATION OF THE VIRUS AND THE TWO VIRAL GENES THAT ARE CARRIED IN AAV REP AND CAP. IF YOU PUT HELPER PLASMIDS FOR ADINOAND REP AND CAP AND WHAT YOU WANT TO PACKAGE, THE GENOME ENGINEERED TO EXPRESS AN ANTIBODY, YOU CAN GET HIGH TITER VIRUS MADE FROM 293 CELLS. SO WE MADE A COUPLE OF EXPRESSION PLASMIDS, ONE OF WHICH ENCODES LUCIFERASE. WE USE THAT TO TRACE WHAT GOES ON AFTER INJECTION OF THE VIRUS AND ONE OF WHICH EXPRESSES ANTIBODIES AND I SHOW HERE THE CONSTRUCT FOR B-12 WHICH TURNS OUT TO BE THE ANTIBODY WE HAVE SPENT THE MOST TIME ON AND THAT WORKS VERY WELL. AND WE ORIGINALLY PUT THESE INTO WILSON'S CONSTRUCTS I WOULD POINT OUT OTHERS HAVE USED A SELF-COMPLIMENTARY AAV. IT GIVES HIGHER TITERS AND SEEMS A BETTER WAY TO GO BUT HALF THE CARRYING CAPACITY OF THE INTACT VIRUS. THAT NOW IS TOO SMALL TO PUT AN ANTIBODY INTO. SO WE'VE GONE WITH THE STANDARD VIRUS RATHER THAN THE WHAT ARE CALLED SOUTH COMPLIMENTARY AAV. WE PRODUCE THEM BY TRANSFECTION, WE SEE THEM IN PART AND PURIFY THEM. THAT GENERATES OVER 10 TO THE 13 GENOME COPIES PER ML IN VIRAL STOCK. SO YOU CAN GET EXTREMELY HIGH TITER STOCKS RELATIVELY EASILY. IF WE DO THIS WITH WITH LUCIFERASE, IF YOU DO AN INTRAVENOUS INJECTION AND IMAGE ANIMALS WHERE LUCIFERASE IS EXPRESSED, IT'S THE LONGNESS OF THE LIVER, THERE'S A LITTLE BIT IN OTHER PLACES, AND THE AMOUNT OF EXPRESSION IS VERY LOW. FEW DO THE SAME THING, TAKE THE SAME PREPARATION AND INJECT IT INTO MUSCLE THIS IS A COLOR LOG SCALE, YOU GET HIGHER EXPRESSION THAT STAYS LOCALIZED TO THE THE LIMB WHERE IT WAS INJECTED. YOU CAN FOLLOW ANIMALS LIKE THAT OUT IN THIS CASE 64 WEEKS, OVER A YEAR AND THE LEVEL OF EXPRESSION DOESN'T VARY A LOT. AND THIS IS THE KEY TO AAV. ONCE IT SETS UP A PRODUCTION OF LUCIFERASE IT MAINTAINS IT OVER A PERIOD OF TIME, IT HAS TO GET INTO A CELL WHICH ITSELF DOESN'T RENEW F. THE CELL IS SELF-RENEWING IT WILL DILUTE OUT THE AAV PLASMID. BUT PARTICULARLY IN MUSCLE, MUSCLE DOESN'T HAVE MUCH CELL TURN OVER, IT'S INTERSTITIAL. SO MUSCLE IS A GREAT PLACE TO MAKE PRODUCT. AND WE'RE NOT THE ONES TO DISCOVER THAT, OTHERS HAVE DISCOVERED THAT. SO WE REPLACED THE LUCIFERASE GENE WITH AN ANTIBODY GENE AND PUT THAT IN AN FOLLOWED THE PRODUCTION OF IGG, HUMAN IGG. IN MICE AND YOU CAN SEE IN THE LOWER CURVE HERE THAT PRK PRODUCTION OF ANTIBODY IS STABLE OVER TIME. THE ORIGINAL CONSTRUCTS WE HAD PUTTING IN 10 TO THE 10 OR 10 TO THE 11th VIRUS THAT'S WHAT THE TWO CURVE THERE IS ARE, THE PRODUCTION OF ANTIBODY MEASURED BY ELISA IN THE BLOOD OF THE ANIMALS WAS -- IS MEASURED IN NANOGRAMS -- SORRY, YES. IN NANOGRAMS PER ML. SO BETWEEN ONE IN TEN MICROGRAMS PER ML. WE DECIDED TO TRY TO IMPROVE THE VECTOR AND WE HAVE DONE A LOT OF THINGS I WON'T GO INTO EVERYTHING, OPTIMIZING THE INSERTS, WE MADE OUR OWN PROMOTER, CALLED A CASSI PROMOTER WHICH IS MORE EFFECTIVE THAN THE CMV PROMOTER THAT IS MORE TRADITIONAL. WE PUT IN THE WPRE, WHICH IS A SORT OF MYTHICAL POST TRANSCRIPTIONAL REGULATORY IMPROVER THAT COMES FROM A WOODCHUCK VIRUS AND GIVES YOU BIG EFFECT USED IN LENTIVIRUSES ALL THE TIME. AND WE AS I SAID, WE IMPROVED THE INSERTS AND CHANGED THE POLYVENTILATION SIGNAL. NOW WE GET HUNDREDS OF MICROGRAMS PER ML OF B-12 STABLY IN THE BLOOD OF ANIMALS OVER A LONG PERIOD OF TIME. THIS EXPERIMENT GOES OUT 7 WEEKS, THE LONGEST WE HAVE GONE IS HALF A YEAR. HERE WE HAVE ACTUALLY FOUR DIFFERENT ANTIBODIES BEING EXPRESSED HERE, B-12, 4E-10 AND 2F-5. 2G 12 AND 4F 5 ARE THE LOWER CURVESCH WE CAN PROBABLY DO BETTER THAN THIS. WE'RE LEARNING MORE ABOUT HOW TO OPTIMIZE EXPRESSION EVERY TIME WE MAKE A NEW CONSTRUCT AND I'M CONVINCED WE CAN GET ANY CONSTRUCT UP INTO THE HUNDREDS OF MICROGRAMS. AND WE'RE NOW GETTING B-12 AT A LEVEL OF A MILLIGRAM PER ML IN THE BLOOD OF MICE. ALL THIS YOU MIGHT SAY WAS DONE WITH IMMUNODEFICIENT ANIMALS BECAUSE WE WANTED TO BE ABLE TO PUT THE HUMAN IMMUNE CELLS INTO THE IMMUNODEFICIENT ANIMALS TO TEST THE EFFECT ON HIV. BUT WE CHECK TO MAKE SURE YOU CAN DO THIS WITH IMMUNOCOMPETENT ANIMALS AND YOU CAN PARTICULARLY IN THE BOTTOM TWO SETS, THE BOB C AND FEB THAT IT WORKS TERRIFICALLY WELL. BHAPS IS THE FIRST WEEK WHEN YOU INJECT THE VIRUS, IT DISTRIBUTES FAIRLY BROADLY ALTHOUGH OBVIOUSLY AT THE SITE OF INJECTION. BUT THEN THE OTHER SITE SLOWLY GO AWAY IN AN IMMUNOCOMPETENT ANIMAL, PROBABLY DUE TO AN IMMUNE REACTION. EXCEPT FOR THE SITE OF THE INJECTION IN THE MUSCLE WHICH STAYS FOR 12 WEEKS AND MORE. I SHOULD POINT OUT THE B-6 ANIMAL WAS BLACK AND THEREFORE YOU CAN'T USE THE XENOGEN WELL ENOUGH TO SEE IT SO FORGET THE MIDDLE CURVE. SO YOU CAN COMPARE THE IMMUNOINCOMPETENT ANIMALS TO THE COMPETENT ANIMALS AND SEE THE DIFFERENCE. YOU CAN FOLLOW LUCIFERASE OUT IN THIS CASE 24 WEEKS AND THE IMMUNOCORPMIZED ANIMAL VERSUS HIGHER -- COMPROMISED ANIMALS HAVE HIGHER OVEREXPRESSION BUT THE COMPETENT ANIMALS MAINTAIN GOOD EXPRESSION. HOWEVER, WHEN WE LOOK AT ANTIBODY RATHER THAN LUCIFERASE, WHAT WE SEE IS VERY LITTLE DIFFERENCE BETWEEN IMMUNOCOMPETENT AND IMMUNOINCOMPETENT ANIMALS. SO WE'RE GETTING REALLY REMARKABLE EXPRESSION OF ANTIBODY, WHETHER THE ANIMALS ARE IMMUNOCOMPETENT OR NOT. SO THE QUESTION THEN IS, DOES THIS ANTIBODY PROTECT AGAINST AN HIV INFECTION? IT CERTAINLY OUGHT TO AT THESE LEVELS. AND IT DOES. WE HAVE DESIGNED NOW A RELATIVELY SIMPLE SYSTEM FOR TESTING THIS. WE USE THE MORE COMPLICATED HUMANIZED MOUSE MODELS FOR SOME TIME. AND FOUND THEM CUMBERSOME, UNREPRODUCIBLE, EXPENSIVE, EVERYTHING WAS WRONG. AND THOUGH THERE ARE SOME ADVANTAGES YOU GET BETTER RECONSTITUTION OF MUCOSAL IMMUNITY, FOR INSTANCE. AND SO FOR CERTAIN KINDS OF THINGS WE STILL USE THE FANCIER HUMANIZED MICE. BUT WHAT WE HAVE BEEN DOING RECENTLY IS A SIMPLE PROTOCOL WHICH WE TAKE AN NSG MOUSE YOU CAN BUY OFF THE SHELF, NOD, SKID, GAMMA, C NEGATIVE. AND INJECT INTO A HUMAN PBMCs. A COUPLE OF WEEKS FOR THE CELLS TO GROW UP, WE INJECT A SMALL NUMBER AND THEY GROW NICELY IN THE ANIMAL AND WE CHALLENGE WITH HIV AND SAMPLE WEEKLY LOOKING IN PARTICULAR AT CD4 CD8 RATIOS IN THE PERIPHERY. BECAUSE THAT'S A VERY QUICK AND GOOD MEASURE. WHAT YOU SEE IN AN UNINFECTED ANIMAL, THAT THE CD4 CD8 RATIO GOES UP WITH THE PERCENTAGE OF CD4 CELLS IN THIS CASE GOES UP WITH TIME BECAUSE THERE'S A CONSTANT NATURAL SELECTION FOR CD4 OVER CD8. I DON'T UNDERSTAND IT BUT PROBABLY NOT INTERESTING. IF YOU INFECT AND WE INFECT IN THIS CASE WITH NL-43 VIRUS THAT'S WHAT THE INFECTIONS I SHOW YOU ARE, THEN YOU SEE AFTER THE FIRST WEEK A CRASH OF CD4 CELLS. THAT CAN GO DOWN STO ZERO DEPENDING ON HOW MUCH VIRUS YOU PUT IN. I MIGHT SAY THIS IS A CXCR-4 VIRUS, TROPIC VIRUS, WE DID THAT PURPOSEFULLY BECAUSE IF YOU LOOK IN THE PERIPHERY AND YOU USE AN R-5 VIRUS YOU DON'T GET COMPLETE ELIMINATION OF THE CELLS VERY QUICKLY. YOU SLOWLY DO BUT NOT QUICKLY. AND SO IT'S HARD TO GET A GOOD WIN DOE IN WHICH TO LOOK AT THE EFFECT OF THE ANTIBODY. HERE THE EFFECT OF THE ANTIBODY, IF THE -- SEE IF I CAN -- YEAH. SO THE BLACK LINE HERE IS THE CONTROL, THE LUCIFERASE VIRUS OR WITH SOME IRRELEVANT ANTIBODY, AND THE TOP LINE IS B-12. AND THE OTHER LINES ARE OTHER MONOCLONAL ANTIBODIES. I THINK THAT'S 2G-12, THAT'S 4E 10, THAT'S 2F 5. THERE AT SOMEWHAT LOWER LEVELS, SO IT'S NOT SURPRISING THEY ARE LESS EFFECTIVE. ALL THE ANTIBODIES, HOWEVER, ARE CLEARLY EFFECTIVE. TBU B-12 IS REMARKABLE BECAUSE THAT'S THE SAME CURVE YOU GET IF YOU HAVE AN UNINFECTED ANIMAL. IT'S PROVIDING COMPLETE PROTECTION AGAINST -- I DIDN'T SAY WHAT THE INFECTION IS HERE, BUT IT'S AN INFECTION WITH TENS OF THOUSANDS OF VIRUS, A HIGH-LEVEL INFECTION. WE DID IT PURPOSEFULLY FOR THIS AND THERE SOMEBODY HERE EARLIER SAID, WHICH IS TO GET DATA. BUT I UNDERSTANDING FULL WELL A VAGINAL INFECTION WOULD BE A LOT MORE APPROPRIATE AT LOW CONCENTRATIONS. BUT YOU DON'T NEED IT BECAUSE WHEN YOU SEE THIS DATA YOU KNOW WHERE YOU'RE AT. WE HAVE SEARCHED THROUGH THE B-12 PROTECTED ANIMALS USING HISTOLOGY TO LOOK FOR P-24 POSITIVE CELLS. WE CAN'T FIND ANY. NON-DETECTABLE. ALL THE OTHER ANTIBODIES WE CAN SEE THE VIRUS GROWING IN THESE ANNALS -- INMALLS -- ANIMALS AND OF COURSE IN THE CONTROL. SO WE CAN'T FIND ANY EVIDENCE WE CAN FIND EVIDENCE OF P-24 IN THESE ANIMALS AN WHETHER IT'S ABSOLUTELY STERILIZING IMMUNITY OR VIRTUALLY STERILIZING IMMUNITY, I DON'T KNOW BUT IT'S IMPRESSIVE. SO WE HAVE ESCALATING THE HIV CHAL LENG FROM USING NOW NANOGRAMS OF P-24 AS MEASURE OF HOW MUCH VIRUS WE HAVE FROM 125 NANOGRAMS DOWN TO VERY LOW LEVELS. .1 I THINK. AND LOOKED AT PROTECTION. WHAT YOU CAN SEE THE THAT THE PROTECTION IS BASICALLY INDEPENDENT OF THE AMOUNT OF VIRUS THAT'S PUT IN. THERE REALLY IS NO DIFFERENCE BETWEEN ANY OF THOSE BLUE CURVES. SO WE ARE NOW CONTINUING THESE EXPERIMENTS, THEY TAKE A LONG TIME I MUST SAY, TO DETERMINE THE SERUM CONCENTRATION OF ANTIBODY THAT WILL DO THIS. WE'RE DOING THAT WITH PG-9, PG-16, VRCO-1, WITH A 2G 12, AN OPTIMIZED WITH OTHER CONSTRUCTS THAT ARE BEING WORKED ON IN PAMELA BJORKLAND'S LAB AND LOOK AT EFFECTS OF IGG VERSUS IGA AND AFTER ASHLEY HAASE'S PRESENTATION, I WANT TO DO THAT EVEN MORE. YOU CAN GET NICE DIFFERENT LEVELS OF ANTIBODY BY PUTTING IN DIFFERENT AMOUNTS OF VIRUS. NOT AN EXACT PROPORTIONALITY BUT IT GIVES YOU DIFFERENT AMOUNTS OF ANTIBODY TO LOOK AT OF PROTECTION ABILITY. THESE B-12 PRODUCING ANIMALS ARE BEING CHALLENGED RIGHT NOW. AND THE PRELIMINARY DATA IS LOW IN THE RANGE OF A FEW MICROGRAMS SEEMS TO BE GIVING PROTECTION. WE HAVE DONE THE SAME WITH VRC-1, AGAIN, IT CLEARLY IS PROTECTIVE BUT HOW IT COMPARES TO B-12 BEDON'T KNOW QUANTITATIVELY AND WON'T KNOW FOR A FEW WEEKS. WE CAN MAKE PG-9 AND 16. THEY NEED OPTIMIZATION. WE KNOW HOW TO DO THAT NOW. OUR FEELING IS, WE KNOW ENOUGH TODAY TO BE CONFIDENT THAT IF HUMANS ACT LIKE MICE, WE CAN PROTECT HUMANS AGAINST HIV INFECTION. WE WANT TO KNOW A LOT OF DETAILS BUT THEY ARE DETAILS. AND WE NEED TO ASK IF HUMANS DO ACT LIKE MICE, DO THEY MAKE PROPHYLACTIC LEVELS OF ANTIBODY EXCUSING AAV AS A VEHICLE FOR PRODUCTION. WE NEED TO INVESTIGATE SAFETY ISSUES BUT I WANT TO POINT OUT THERAPEUTIC ANTIBODIES HAVE PROVED TO BE QUITE SAFE IN MANY DIFFERENT SETTINGS NOW BUT IT WILL REQUIRE CLINICAL TRIALS TO KNOW WHETHER THIS FORM OF PROTECTION CAN BE TAKEN TO HUPS OR NOT. THANK YOU VERY MUCH. [APPLAUSE] I'LL LEAVE THOSE LAST TWO. >> I WANT TO THANK EVERYBODY FOR JOINING US TODAY AND I AM HOPEFUL THAT BY THE TIME OF THE 15th YEAR REUNION WE'LL BE TALKING ABOUT THE VACCINE. I LOOK FORWARD TO JOINING SOME OF YOU OVER IN THE STONE HOUSE AFTERWARDS FOR THE RECEPTION. THANK YOU. THANK YOU.