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Biological and immunological characterization of plant-produced HIV-1 Gag/dgp41 virus-like particles
Description
Anti-retroviral drugs and AIDS prevention programs have helped to decrease the rate of new HIV-1 infections in some communities, however, a prophylactic vaccine is still needed to control the epidemic world-wide. Despite over two decades of research, a vaccine against HIV-1 remains elusive, although recent clinical trials have shown promising results. Recent successes have focused on highly conserved, mucosally-targeted antigens within HIV-1 such as the membrane proximal external region (MPER) of the envelope protein, gp41. MPER has been shown to play critical roles in the viral mucosal transmission, though this peptide is not immunogenic on its own. Gag is a structural protein configuring the enveloped virus particles, and has been suggested to constitute a target of the cellular immunity potentially controlling the viral load. It was hypothesized that HIV-1 enveloped virus-like particles (VLPs) consisting of Gag and a deconstructed form of gp41 comprising the MPER, transmembrane, and cytoplasmic domains (dgp41) could be expressed in plants. Plant-optimized HIV-1 genes were constructed and expressed in Nicotiana benthamiana by stable transformation, or transiently using a tobacco mosaic virus-based expression system or a combination of both. Results of biophysical, biochemical and electron microscopy characterization demonstrated that plant cells could support not only the formation of HIV-1 Gag VLPs, but also the accumulation of VLPs that incorporated dgp41. These particles were purified and utilized in mice immunization experiments. Prime-boost strategies combining systemic and mucosal priming with systemic boosting using two different vaccine candidates (VLPs and CTB-MPR - a fusion of MPER and the B-subunit of cholera toxin) were administered to BALB/c mice. Serum antibody responses against both the Gag and gp41 antigens could be elicited in mice systemically primed with VLPs and these responses could be recalled following systemic boosting with VLPs. In addition, mucosal priming with VLPs allowed for a robust boosting response against Gag and gp41 when boosted with either candidate. Functional assays of these antibodies are in progress to test the antibodies' effectiveness in neutralizing and preventing mucosal transmission of HIV-1. This immunogenicity of plant-based Gag/dgp41 VLPs represents an important milestone on the road towards a broadly-efficacious and inexpensive subunit vaccine against HIV-1.
ContributorsKessans, Sarah (Author) / Mor, Tsafrir S (Thesis advisor) / Matoba, Nobuyuki (Committee member) / Mason, Hugh (Committee member) / Hogue, Brenda (Committee member) / Fromme, Petra (Committee member) / Arizona State University (Publisher)
Created2011
ContributorsASU Library. Music Library (Publisher)
Created2018-04-09
ContributorsJin, Leon (Performer) / Duo, Hongzuo (Performer) / Bergstedt, David (Performer) / Ellis, Gage (Performer) / Novak, Gail (Performer) / ASU Library. Music Library (Publisher)
Created2021-02-24
ContributorsWaters, Jared (Performer) / Creviston, Hannah (Performer) / Liu, Miao (Performer) / Guo, Hongzuo (Performer) / DeLaCruz, Nathaniel (Performer) / LoGuidice, Rosa (Performer) / Chiko, Ty (Performer) / Gatchel, David (Performer) / ASU Library. Music Library (Publisher)
Created2021-01-28
ContributorsKosminov, Vladislav (Performer) / Solari, John (Performer) / Liu, Shiyu (Performer) / Huang, Anruo (Performer) / Holly, Sean (Performer) / Novak, Gail (Performer) / Yang, Elliot (Performer) / Wu, Selene (Performer) / Kinnard, Zachariah (Performer) / Kuebelbeck, Stephen (Performer) / Johnson, Kaitlyn (Performer) / Bosworth, Robert (Performer) / Matejek, Ryan (Performer) / ASU Library. Music Library (Publisher)
Created2021-01-27
ContributorsSuehiro, Dylan (Conductor) / Kelley, Karen (Performer) / Ladley, Teddy (Performer) / ASU Library. Music Library (Publisher)
Created2021-04-19
Description
In order to cope with the decreasing availability of symphony jobs and collegiate faculty positions, many musicians are starting to pursue less traditional career paths. Also, to combat declining audiences, musicians are exploring ways to cultivate new and enthusiastic listeners through relevant and engaging performances. Due to these challenges, many community-based chamber music ensembles have been formed throughout the United States. These groups not only focus on performing classical music, but serve the needs of their communities as well. The problem, however, is that many musicians have not learned the business skills necessary to create these career opportunities. In this document I discuss the steps ensembles must take to develop sustainable careers. I first analyze how groups build a strong foundation through getting to know their communities and creating core values. I then discuss branding and marketing so ensembles can develop a public image and learn how to publicize themselves. This is followed by an investigation of how ensembles make and organize their money. I then examine the ways groups ensure long-lasting relationships with their communities and within the ensemble. I end by presenting three case studies of professional ensembles to show how groups create and maintain successful careers. Ensembles must develop entrepreneurship skills in addition to cultivating their artistry. These business concepts are crucial to the longevity of chamber groups. Through interviews of successful ensemble members and my own personal experiences in the Tetra String Quartet, I provide a guide for musicians to use when creating a community-based ensemble.
ContributorsDalbey, Jenna (Author) / Landschoot, Thomas (Thesis advisor) / McLin, Katherine (Committee member) / Ryan, Russell (Committee member) / Solis, Theodore (Committee member) / Spring, Robert (Committee member) / Arizona State University (Publisher)
Created2013