David B. Weiner, Ph.D. - US grants

Traditionally, immune therapeutic vaccination against viruses has utilized either live attenuated preparations, whole killed virus or recombinant proteins. Of these approaches, only live attenuated virus preparations activate all arms if the immune system in a manner similar to infection. Genetic immunization is dependent upon injection of a nucleic acid, sequence directly into a host target tissue. In theory, direct genetic immunization would mimic aspects of attenuated vaccines in that synthesis of specific foreign proteins would be accomplished in the host and become the subject of immune surveillance via both the MHC class I and class II pathways. The use of this new technology to immunize animals with human immunodeficiency virus type 1 (HIV-1) envelope gp160 DNA constructs and achieve relevant immune responses has been reported by this group. Antisera from genetically immunized animals including, mice, rabbits and non-human primates, contains anti-HIV envelope glycoprotein immune responses. The antiserum neutralizes HIV-1 infection and inhibits cell to cell infection in vitro. This technology induced both T cell proliferation and isotype switching consistent with the production of relevant T helper immune responses. Lysis of relevant env expressing targets was induced by immune spleenocytes from mice and primates. Through the development of an in vivo murine model it was demonstrated that mice can reject lethal cell challenge through specific immune responses directed at HIV proteins. This in vivo anti HOV data suggest the utility of this technology for anti-HIV immmune therapeutic strategies. This grant proposal will focus on the safety of this strategy and its ability to effectively induce immune responses in humans and non-human primates previously infected with HIV or SIV. Project 1 will evaluate the safety, immunogenicity and efficacy of simian immunodeficiency virus (SIV) constructs encoding gag/pol or env given singly or in combination to rhesus macques infected with SIV. Project 2 will evaluate the safety, immunogenicity and efficacy of genetic immunization with human immunodeficiency virus (HIV) constructs encoding gag/pol or env given singly or in combination to chimpanzees infected with HIV. Project 3 will evaluate the safety, immunogenicity and responses to genetic immunization with HIV constructs encoding gag/pol or env given singly or in combination in HIV+humans. A molecular core will be responsible for development of clinical grade material for genetic immunization. An administrative core will coordinate the interactions and sample handling. These studies will generate critical data regarding athe safety, immunogenicity and utility of genetic inoculation with HIV-1 genes to induce immune responses as potential immune therapy for HIV-1.

Traditionally, vaccination against pathogenic human viruses has utilized either live attenuated preparations, whole killed virus or recombinant proteins. Of these approaches, only live attenuated virus preparations activate all arms of the immune system in a manner similar to native infection. Genetic immunization is dependent upon injection of a nucleic acid sequence directly into a host target tissue. In theory, direct genetic immunization should mimic aspects of attenuated vaccines in that synthesis of specific foreign proteins would be accomplished in the host and become the subject of immune surveillance via both the MHC class I and class II pathways. The use of this new technology to immunize animals with a human immunodeficiency virus type 1 (HIV-1) envelope gp 160 DNA construct and achieve relevant immune responses has been reported by this group. Antisera from genetically immunized animals including, mice, rabbits and non-human primates, demonstrate anti-HIV envelope glycoprotein immune responses. The antiserum neutralizes HIV-1 infection and inhibits cell to cell infection in vitro. This technology induced both T cell proliferation and isotype switching consistent with the production of relevant T helper immune responses. Furthermore it was demonstrated that CTL lysis of relevant env expressing targets was similarly induced. Through the development of an in vivo murine model we demonstrated that mice can reject lethal cell challenge through specific immune responses directed at HIV proteins. This in vivo anti HIV data suggest the utility of this specific immune responses elicited by intramuscular injection of genes. While all arms of the immune system are elicited by genetic immunization, the fine specificity of the responses induced have not been characterized in detail. More importantly, the ability of this strategy to induce protective responses in humans and non-human primates has yet to be established. This proposal will focus on the analysis of this approach to effectively induce such protective immunity from lentiviral challenge. The overall strategy will be to perform in depth analysis of specific immune responses against SIV constructs in mice, which will aid our studies in non-human primates. Constructs will be developed for expression of the SIV239 env and gag/pol genes and evaluated in vitro for expression in relevant cell lines. These will then be analyzed in preliminary inoculation studies in mice, their in vivo immunogenicity will be analyzed. These results will give insight into the experiments in non- human primates where similar analysis of the immune responses generated by construct inoculation will be performed. Finally the SIV constructs will be analyzed for their protective potential in vivo in viral challenge experiments. Together these studies will characterize the potential of this new technology for the development of facilitated genetic inoculation protocols as a potential prophylactic vaccine or against pathogenic primate lentiviruses. These studies have direct relevance to vaccine development against HIV-1.

Traditionally, immune therapeutic vaccination against viruses has utilized either live attenuated preparations, whole killed virus or recombinant proteins. Of these approaches, only live attenuated virus preparations activate all arms of the immune system in a manner similar to infection. Genetic immunization is dependent upon injection of a nucleic acid sequence directly into a host target tissue. In theory, direct genetic immunization should mimic aspects of attenuated vaccines in that synthesis of specific foreign proteins would be accomplished in the host and become the subject of immune surveillance via both the MHC class I and class II pathways. The use of this new technology to immunize animals with human immunodeficiency virus type 1 (HIV-1) envelope gp 160 DNA constructs and achieve relevant immune responses has been reported by this group. Antisera from genetically immunized animals including, mice, rabbits and non-human primates, contains anti-HIV envelope glycoprotein immune responses. The antiserum neutralizes HIV 1 infection and inhibits cell to cell infection in vitro. This technology induced both T cell proliferation and isotype switching consistent with the production of relevant T helper immune responses. Heterologous lysis of relevant env expressing targets was induced by immune spleenocytes from mice and primates. This in vivo anti HIV-1 data support the utility of this technology for anti- HIV immune therapeutic strategies. The chimpanzee is the prototypic model for HIV infection studies. The goal of this project is to evaluate the safety and efficacy of genetic inoculation with human immunodeficiency virus (HIV) constructs encoding gap/pol or env given singly or in combination in chimpanzees infected with HIV-1. Additionally, the analysis of viral load and immune responses in infected animals following genetic inoculation will generate important data regarding the immunogenicity and utility of this approach as a therapeutic strategy for HIV-1 infection. Additionally, this project will focus on the detailed analysis of humoral and cellular immune responses of the patients participating in project 3 and for the SIV studies in project 1.

Traditionally, immune therapeutic vaccination against viruses has utilized either live attenuated preparations, whole killed virus or recombinant proteins. Of these approaches, only live attenuated virus preparations activate all arms if the immune system in a manner similar to infection. Genetic immunization is dependent upon injection of a nucleic acid, sequence directly into a host target tissue. In theory, direct genetic immunization would mimic aspects of attenuated vaccines in that synthesis of specific foreign proteins would be accomplished in the host and become the subject of immune surveillance via both the MHC class I and class II pathways. The use of this new technology to immunize animals with human immunodeficiency virus type 1 (HIV-1) envelope gp160 DNA constructs and achieve relevant immune responses has been reported by this group. Antisera from genetically immunized animals including, mice, rabbits and non-human primates, contains anti-HIV envelope glycoprotein immune responses. The antiserum neutralizes HIV-1 infection and inhibits cell to cell infection in vitro. This technology induced both T cell proliferation and isotype switching consistent with the production of relevant T helper immune responses. Lysis of relevant env expressing targets was induced by immune spleenocytes from mice and primates. Through the development of an in vivo murine model it was demonstrated that mice can reject lethal cell challenge through specific immune responses directed at HIV proteins. This in vivo anti HOV data suggest the utility of this technology for anti-HIV immmune therapeutic strategies. This grant proposal will focus on the safety of this strategy and its ability to effectively induce immune responses in humans and non-human primates previously infected with HIV or SIV. Project 1 will evaluate the safety, immunogenicity and efficacy of simian immunodeficiency virus (SIV) constructs encoding gag/pol or env given singly or in combination to rhesus macques infected with SIV. Project 2 will evaluate the safety, immunogenicity and efficacy of genetic immunization with human immunodeficiency virus (HIV) constructs encoding gag/pol or env given singly or in combination to chimpanzees infected with HIV. Project 3 will evaluate the safety, immunogenicity and responses to genetic immunization with HIV constructs encoding gag/pol or env given singly or in combination in HIV+humans. A molecular core will be responsible for development of clinical grade material for genetic immunization. An administrative core will coordinate the interactions and sample handling. These studies will generate critical data regarding athe safety, immunogenicity and utility of genetic inoculation with HIV-1 genes to induce immune responses as potential immune therapy for HIV-1.

DNA vaccines engineered to elicit immune responses against HIV-1 can induce humoral and cellular immune responses in a variety of species. We have reported on the development of a multi-cassette system for the production of broad-based immune responses in vivo against the majority of HIV-1 open reading frame encoded antigens. Our group has shown that through a multiple immunization regime this approach can impact on viral load in primate models of infection. Specifically, we have shown that DNA vaccine constructs can protect chimpanzees from HIV challenge, can protect a subset of macaques from SHIV challenge as well as appear to function as therapeutic agents to lower viral load in infected chimpanzees. Preliminary studies demonstrate that HIV-1 DNA vaccines are apparently well tolerated and are immunogenic in humans (see Administrative core and overview, Project 3 and Immunology Core). However, it is unclear the degree to which the type of immunity induced will be capable of impacting on HIV infection in a natural setting using these first generation vaccines. Furthermore the multiple immunization scheme we used previously in the chimpanzee model might be cumbersome for testing of these vaccine candidates. In this regard, it may be important to specifically engineer immune responses in order to improve on first generation vaccine attempts. Especially for HIV, induction of cell-mediated immunity may be an important feature for any candidate vaccine as such responses could interrupt the establishment of infection. The humoral infection response may play a role in limiting viral entry and slowing viral spread during the infection process. Furthermore the induction of chemokines in such immunization procedures may be important for limiting viral entry and infection (see Project 2). This project will attempt to engineer enhancement of in vivo cellular and humoral immune responses to HIV-1 DNA vaccine cassettes through the use of immunomodulatory gene constructs. A panel of immunomodulatory genes will be studied in a system which we have designed and exploited Studies will be performed in mice, and both macaques and chimpanzees to characterize and optimize immunogenicity of these novel constructs. The effects of these vaccine candidates to impact viral load in challenge studies will be monitored. These studies will allow us to develop a new generation of HIV-DNA vaccine for testing in humans (see Project 3). Additionally, these studies are designed to enhance our understanding of the types of immune responses which are correlated with any observed protection by not just observing and cataloguing induced responses but on the front end driving responses in a particular immunological direction.

A DNA-based vaccine for HIV-1 will have to deal with the diversity of this virus, as well as issues of potency and effectiveness. This proposal is part of a large program undertaken by this research group aimed to exploit the utility of DNA to ultimately develop a prophylactic vaccine for HIV-1. We have designed a prototypic DNA based vaccine for HIV-1 which includes attenuated gag/pol genes, attenuated accessory genes, a prototypic env/rev immunization cassette and is likely to require additional envelope subtypes in an effort to deal with viral diversity and to limit viral escape. Furthermore one question central question regarding this novel technology is its potency. Accordingly, this proposal will focus on the generation of improved DNA immunogens through engineering gene expression cassettes which co-deliver cytokine as well as other immunomodulatory gene sequences as part of the vaccine. We will directly test this concept in mice and in primates and in clinical studies. There are three projects and two cores which form this proposal. Project 1 will develop and test cytokines and co-stimulatory molecules as molecular adjuvants for HIV-1 immunization cassettes and in particular monitor the cellular effects of such vaccine manipulation. Project 2 will test the serological consequences as well as effects on chemokine production induced by the use of cytokines and co-stimulatory molecules as molecular adjuvants to enhance HIV-1 immunization cassettes. Project 3 will test a combined vaccine cocktail of gag/pol and env/rev vaccine cassettes first and then go on to test a subset of the molecular adjuvant cassettes in clinical studies. These studies will define the initial safety profile as well as test their ability to impact on their specific immune responsiveness in humans. These projects are supported by an experienced Central Immunology core as well as an experienced Molecular Biology core laboratory. The projects and cores and subcontracts are coordinated by the Central Administrative core. This program project brings together a seasoned, collaborative and highly interactive group of investigators who have moved DNA vaccines from concept to the lab bench and to the clinic. In this proposal we will develop the second generation of tools for optimizing this new technology in an attempt to maximize the effectiveness of these vaccines and to probe the relationship of directed immune responses to impact on viral replication in vivo. This program will directly test the ability of such second generation vaccines to impact on HIV replication in animal models and to modulate HIV immune responsiveness in humans by not just characterizing but by driving immune response direction in vivo.

We have moved a DNA vaccine concept to the point of clinical testing through the IPCP funding mechanism. Our data supports that DNA vaccines can be improved by co-delivery of specific plasmid encoded cytokine adjuvants in non-human primates. This program evaluated interesting DNA Vaccine adjuvants in macaques. Results of this preclinical testing identified interleukin-12 (IL-12) and interleukin-15 (IL-15) as the most promising for clinical development. We developed substantial evidence that IL-12 can act as a highly potent DNA vaccine adjuvant in SIV infected primates for driving CTLs that appear to impact viral replication. This therapeutic study in HIV infected individuals, that was included under project 2 of the original IPCP proposal could not be completed due to delays in the manufacturing of the DNA plasmids and the additional time needed to perform the toxicity and biodistribution studies necessary to support the clinical trial. The proposed study design will test the safety and Jmmunogenicity of an optimized multigene HIV-1 DNA immunogen (PENNVAX-B) when used alone and adjuvanted with plasmid IL-12 DNA or plasmid IL-15 DNA in the context of HIV infection. Another parallel trial, conducted by the HVTN will test these products in HIV uninfected individuals.

[unreadable] DESCRIPTION (provided by applicant): The current bioterrorism threat has refocused our nation on the issue of our population's susceptibility to a smallpox attack. As a precaution, deployment of dilution of the current stock of vaccine and deployment of a tissue culture version of the VACV vaccine have been requisitioned. This vaccine and likely the new stocks carried a risk of Significant Adverse Events of 1 per 10,000 vaccinated individuals. However, that vaccine was last used in a very different environment. The high-risk groups for AE's include the elderly, the very young, immunocompromised individuals and others. The percentage and absolute numbers of the US population that falls within these categories has risen dramatically in the past 30 years. The greater than 750,000 persons living in the US that are HIV-positive is one clear example. This suggests that the AE risk of the VACV may be unacceptable and can only be deployed as a last resort. However, the option of abandoning this approach and developing new approaches leaves us at risk for a possibly unknown period of time. The hypothesis to be tested in this application is that there is a third option, to develop a strategy that uses the current vaccine yet limits its pathogenesis while improving its potency. It is our hypothesis that priming with enhanced expressing plasmid vaccines that induce nonneutralizing cellular immune responses will prime for successful and even enhanced boosting with the current vaccine, yet limit its associated pathogenesis. This application will use quantitative T cell assays including Elispot and ICC and tetramer analysis, and novel human HLA+DR positive transgenic mice that we have developed to pursue the three specific aims. Novel cellular reagents including MHC class I tetramers will be developed that will have significant value in following VACV challenge in humans as they may be useful as cellular surrogates for the current site reaction take. Together these studies will establish if this simple and novel approach can bridge the current situation and produce a safer more effective smallpox vaccine. [unreadable] [unreadable]

Based on a multitude of data there are several features desired in an HIV Vaccine immunogen. Such an immunogen should induce strong and broad humoral and cellular immunity. Furthermore, as HIV is in general a sexually transmitted disease and the cells of the gut are preferentially targeted for viral destruction, an immunogen should be capable of inducing in particular mucosal as well as systemic immune responses. Currently there is no approach that can be simply administered that induces such a response. In this regard, Dr. Weiner's laboratory first reported that they could redirect immune cells in vivo using chemokines encoded as part of a DMAvaccine cocktail, and recent work further confirmed and elegantly extended these findings through modification of vaccine induced immune cell trafficking by utilizing chemokines (immune trafficking signals) to attract peripheral immune cell populations. It is now the goal of this application to extend this work and develop a mucosal vaccine strategy that will result in the redirection of cells of the mucosal compartment in response to a DNA vaccine administered in the systemic compartment. Our preliminary data support that this strategy generates features of mucosal immunity by systemic vaccination. This application will further investigate this novel approach in this exceptionally important area of vaccine development. We will study the ability of specific chemokines as DNA vaccine adjuvants to modulate immune cell trafficking and redirect effector T and B cell responses to mucosal sites. There are 4 Specific Aims outlined in Project 1 of this program that will address the following questions: First, will delivery of chemokine immunoadjuvants systemically by DNA vaccines induce antigen specific immune responses in mucosal sites, and secondly, can mucosal-derived chemokine-induced immunogenicity be explained by a mechanism in which chemokines induce "retrafficking" of organ specific homing routes? Alternatively, is it such that the "imprinting" dogma for peripheral/mucosal immune cells, in fact, is reversible, resulting from chemokine-induced activation and "re-education" of target cells displaying new homing potentials. Finally, we will test whether chemokine adjuvants can elicit physiologically relevant cellular and humoral immune responses that can protect mice from a lethal mucosal challenge. This project will also generate all constructs for the macaque studies for Project 2 and support the polyfunctional flow studies in Project 3. These studies have great significance for our basic understanding of lymphocyte homing to the gut, mucosal phenotype commitment and for the development of an HIV vaccine that delivers antigens to generate mucosal immunity.

[unreadable] DESCRIPTION (provided by applicant): In response to the stated NIAID growing need for expertise in the area of vaccine research and development, the medical research community at the University of Pennsylvania proposes to establish a training program in Vaccines and Immune Therapeutics. The University Campus has an important history of contribution to vaccine discovery, development and testing of many currently licensed vaccines and has developed an important program to share this expertise with trainees. Trainers are drawn from the Wistar Institute, University of Pennsylvania, as well as the Children's Hospital of Philadelphia, which are institutions located contiguously on the University of Pennsylvania campus. The training program would initially support 3 students among a pool of Ph.D or MD/Ph.D or V.M.D./Ph.D candidates, as well as 1 post doctoral fellow. This application would allow the trainees to train in one of 21 laboratories directed by highly motivated Principal Investigators who have established records in training in important areas of Vaccine and Immune Therapy research. The trainers associated with this T32 application were selected based on their well established record of accomplishment in Vaccine/Immune Therapy-related research as evidenced by their reputations, strong publication records and grant funding, as well as a strong group history of minority training and recruitment and strong reviews for participation in student training. This training program will be part of the newly defined graduate training program in Gene Therapy and Vaccines (GTV), which is one of the 6 programs in the Cell and Molecular Biology Graduate Program (CAMB). The affiliation with CAMB, the largest biomedical graduate training program at the University of Pennsylvania, will facilitate student training and stability on the campus. Furthermore a unique externship relationship with industry allows students and postdoctoral trainees to experience training at some of the nations elite company vaccine settings in a highly formalized manner. The GTV training program has formal mechanisms to monitor trainees' progress both during and after their training grant support. Specialized courses have been developed and are in place to ensure significant grounding in Vaccines and Immune Therapy. An exceptional group of faculty has been assembled to assist in this process. Importantly, the training program has formal mechanisms to monitor trainers and to ensure a high level of accomplishment for trainees and faculty alike. [unreadable] [unreadable] [unreadable]

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DESCRIPTION (provided by applicant): There is a great need to develop new vaccination approaches for influenza. The current strategy based on waiting to identify each seasons new strains and then building a matching vaccine based mostly on egg production is not ideal. Because of this requirement to match strains and the limits on egg based production, the US is forced to under vaccinate our population due to limitations in supply and the impossible task of vaccinating the entire US population each year. This application proposes a new vaccine approach that if successful can result in a transformative shift in the development of seasonal influenza vaccines. We propose to develop a synthetic collection of HAs encoding a focused set of consensus H1, H3 & HB immunogens as a cocktail that together produce broad HI and uNeutralization against seasonal and pandemic H1, H3 and HB viruses. We use an improved DNA technology which eliminates virus culture entirely. We have assembled an outstanding team of investigators who together have helped to produce the technology and the advances that underpin this important application. There are four specific aims that make up this application. Aim 1: To test the hypothesis that we can create a synthetic H1HA immunogen which generates broad HI antibody responses against all four H1 pandemic strains as well against the last 20 years seasonal H1 viruses in mice and ferret models. Aim 2: To test the hypothesis that we can create a synthetic H3HA as well as HBHA which will each generate broad HI responses against seasonal variants of H3 and HB viruses in mice and ferret models. Aim 3: To test the hypothesis that we can develop a combination immunization strategy for H1, H3 and HB immunogens resulting in broad HI activity at protective levels (greater than 1/40 HI titers in an indicator animal species - Ferrets). Studies in ferrets will include challenge with pandemic as well as seasonal strains. We will confirm the immune potency of this combination vaccine in the Rhesus Macaque model which will provide further evidence of the immune potency of the optimized enhanced delivery DNA platform. Aim 4: To test the hypothesis that we can safely induce, using a well tolerated platform, protective HI levels in humans in a pilot safety and immunogenicity study using the combination H1, H3, HB universal DNA vaccine when delivered to healthy volunteers. Together these aims represent transformational shifts for the development of vaccines to influenza as well as for the DNA vaccine field as a whole.

DESCRIPTION (provided by applicant): In response to the stated NIAID growing need for expertise in the area of vaccine research and development, the medical research community at the University of Pennsylvania proposes to establish a training program in Vaccines and Immune Therapeutics. The University Campus has an important history of contribution to vaccine discovery, development and testing of many currently licensed vaccines and has developed an important program to share this expertise with trainees. Trainers are drawn from the Wistar Institute, University of Pennsylvania, as well as the Children's Hospital of Philadelphia, which are institutions located contiguously on the University of Pennsylvania campus. The training program would initially support 3 students among a pool of Ph.D or MD/Ph.D or V.M.D./Ph.D candidates, as well as 1 post doctoral fellow. This application would allow the trainees to train in one of 21 laboratories directed by highly motivated Principal Investigators who have established records in training in important areas of Vaccine and Immune Therapy research. The trainers associated with this T32 application were selected based on their well established record of accomplishment in Vaccine/Immune Therapy-related research as evidenced by their reputations, strong publication records and grant funding, as well as a strong group history of minority training and recruitment and strong reviews for participation in student training. This training program will be part of the newly defined graduate training program in Gene Therapy and Vaccines (GTV), which is one of the 6 programs in the Cell and Molecular Biology Graduate Program (CAMB). The affiliation with CAMB, the largest biomedical graduate training program at the University of Pennsylvania, will facilitate student training and stability on the campus. Furthermore a unique externship relationship with industry allows students and postdoctoral trainees to experience training at some of the nations elite company vaccine settings in a highly formalized manner. The GTV training program has formal mechanisms to monitor trainees' progress both during and after their training grant support. Specialized courses have been developed and are in place to ensure significant grounding in Vaccines and Immune Therapy. An exceptional group of faculty has been assembled to assist in this process. Importantly, the training program has formal mechanisms to monitor trainers and to ensure a high level of accomplishment for trainees and faculty alike.

Project 1 Abstract An effective HIV vaccine remains an elusive goal. The possible limited success of the RV144 vaccine trial supports that it is possible to provide protection from HIV acquisition through immunization. However, the vaccine-induced responses elicited by the RV144 ALVAC vCP1521-AIDSVAX B/E strategy were modest and of limited durability. The immune responses induced in a preferred vaccine should be superior to those observed in RV-144 (low T cells, limted antibodies, partially effective) as well as STEP and HVTN 505 (limited antibody responses, non effective T cell responses). A vaccine platform that avoids induction of anti-vector immunity is of growing importance in light of possible enhancement issues. Currently, there is no such HIV vaccine available, and few groups have the combination of technologies proven in the clinic to produce such a platform. This innovative program makes major advances in new DNA adaptive EP + gene adjvuant vaccine technology which in the clinic generates T cell immunity equivalent or superior to live viral vector vaccines (30, 2). We will build on this recent clinical success by novel genetic adjuvants focused on improved antibody induction as well as next generation adaptive EP focusing on skin delivery. We concentrate on increasing the breadth of coverage induced by these designed DNA vaccine by exploring the potential of a multivalent DNA prime followed by a multivalent protein boost. Furthermore, we plan to develop this collection of technologies in a simplified vaccine scheme that has distinct clinical advantages for global testing. There are three aims that comprise this program.

Abstract Administrative Core - IPCAVD -? E-DNA + Env protein Vaccination for HIV The goal of this program to produce an enhanced E/P delivered gene adjuvanted DNA vaccine (E-DNA) and combine this with a highly novel polyvalent A, B, C, A/E recombinant protein boost (PPB) as a vaccine platform. The EP focus of this application is to utilize novel surface delivery. By this combination we hypothesize that we will generate an improved spectrum of T and B anti HIV immune responses compared to current HIV vaccine modalities. This project builds on a major innovations and accomplishments by the members of this team that have led to this proposal. The team has worked together productively for multiple years. Furthermore they have an outstanding track record of productivity working with the HVTN. The inclusion of CHAVI investigators as part of the preclinical development immune analysis is a major strength of the program. There are 3 highly interrelated, translational focused, exceptionally novel projects which build on a strong track record of accomplishment together that comprise this program. They are supported by a highly respected, important protein core as well as seasoned administrative core. The Administrative core (AC) is responsible for overall program performance, sample distribution, programmatic oversight, HVTN and NIH Programmatic interactions and for recommending SAB members for program review. The AC will arrange the annual SAB meeting and put in place a work plan to act on the SAB?s suggestions with guidance of NIH program. The external SAB will be put in place upon notification of award of this program. The administrative core will arrange team calls for open data discussion, and time line review, arrange monthly calls with NIH program, arrange monthly calls to review progress with the PI and teams, arrange biannual data review meetings and provide yearly written reports on progress for program feedback as well as facilitate publication of findings and scientific meeting attendance for result reporting. The AC will also update the HVTN on a quarterly basis relative to programmatic goals. We have an established HVTN development team to facilitate this process and this will be expanded based on this application being funded (HVTN letter).