ASU Electronic Theses and Dissertations
This collection includes most of the ASU Theses and Dissertations from 2011 to present. ASU Theses and Dissertations are available in downloadable PDF format; however, a small percentage of items are under embargo. Information about the dissertations/theses includes degree information, committee members, an abstract, supporting data or media.
In addition to the electronic theses found in the ASU Digital Repository, ASU Theses and Dissertations can be found in the ASU Library Catalog.
Dissertations and Theses granted by Arizona State University are archived and made available through a joint effort of the ASU Graduate College and the ASU Libraries. For more information or questions about this collection contact or visit the Digital Repository ETD Library Guide or contact the ASU Graduate College at gradformat@asu.edu.
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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
Description
Functional traits research has improved our understanding of how plants respond to their environments, identifying key trade-offs among traits. These studies primarily rely on correlative methods to infer trade-offs and often overlook traits that are difficult to measure (e.g., root traits, tissue senescence rates), limiting their predictive ability under novel conditions. I aimed to address these limitations and develop a better understanding of the trait space occupied by trees by integrating data and process models, spanning leaves to whole-trees, via modern statistical and computational methods. My first research chapter (Chapter 2) simultaneously fits a photosynthesis model to measurements of fluorescence and photosynthetic response curves, improving estimates of mesophyll conductance (gm) and other photosynthetic traits. I assessed how gm varies across environmental gradients and relates to other photosynthetic traits for 4 woody species in Arizona. I found that gm was lower at high aridity sites, varied little within a site, and is an important trait for obtaining accurate estimates of photosynthesis and related traits under dry conditions. Chapter 3 evaluates the importance of functional traits for whole-tree performance by fitting an individual-based model of tree growth and mortality to millions of measurements of tree heights and diameters to assess the theoretical trait space (TTS) of “healthy” North American trees. The TTS contained complicated, multi-variate structure indicative of potential trade-offs leading to successful growth. In Chapter 4, I applied an environmental filter (light stress) to the TTS, leading to simulated stand-level mortality rates up to 50%. Tree-level mortality was explained by 6 of the 32 traits explored, with the most important being radiation-use efficiency. The multidimentional space comprising these 6 traits differed in volume and location between trees that survived and died, indicating that selective mortality alters the TTS.
ContributorsFell, Michael (Author) / Ogle, Kiona (Thesis advisor) / Barber, Jarrett (Committee member) / Hultine, Kevin (Committee member) / Franklin, Janet (Committee member) / Day, Thomas (Committee member) / Arizona State University (Publisher)
Created2017
Description
Throughout the Southwest, complex geology and physiography concomitant with climatic variability contribute to diverse stream hydrogeomorphologies. Many riparian plant species store their seeds in soil seed banks, and germinate in response to moisture pulses, but the climatic controls of this response are poorly understood. To better understand the ecological implications of a changing climate on riparian plant communities, I investigated seed bank responses to seasonal temperature patterns and to stream hydrogeomorphic type. I asked the following questions: Are there distinct suites of warm and cool temperature germinating species associated with Southwestern streams; how do they differ between riparian and terrestrial zones, and between ephemeral and perennial streams? How does alpha diversity of the soil seed bank differ between streams with ephemeral, intermittent, and perennial flow, and between montane and basin streams? Do streams with greater elevational change have higher riparian zone seed bank beta-diversity? Does nestedness or turnover contribute more to within stream beta-diversity?
I collected soil samples from the riparian and terrestrial zones of 21 sites, placing them in growth chambers at one of two temperature regimes, and monitoring emergence of seedlings for 12 weeks. Results showed an approximately equal number of warm and cool specialists in both riparian and terrestrials zones; generalists also were abundant, particularly in the riparian zone. The number of temperature specialists and generalists in the riparian zones did not differ significantly between perennial headwater and ephemeral stream types. In montane streams, alpha diversity of the soil seed bank was highest for ephemeral reaches; in basin streams the intermittent and perennial reaches had higher diversity. Spatial turnover was primarily responsible for within stream beta-diversity—reaches had different species assemblages. The large portion of temperature specialists found in riparian seed banks indicates that even with available moisture riparian zone plant community composition will likely be impacted by changing temperatures. However, the presence of so many temperature generalists in the riparian zones suggests that some component of the seed bank is adapted to variable conditions and might offer resilience in a changing climate. Study results confirm the importance of conserving multiple hydrogeomorphic reach types because they support unique species assemblages.
I collected soil samples from the riparian and terrestrial zones of 21 sites, placing them in growth chambers at one of two temperature regimes, and monitoring emergence of seedlings for 12 weeks. Results showed an approximately equal number of warm and cool specialists in both riparian and terrestrials zones; generalists also were abundant, particularly in the riparian zone. The number of temperature specialists and generalists in the riparian zones did not differ significantly between perennial headwater and ephemeral stream types. In montane streams, alpha diversity of the soil seed bank was highest for ephemeral reaches; in basin streams the intermittent and perennial reaches had higher diversity. Spatial turnover was primarily responsible for within stream beta-diversity—reaches had different species assemblages. The large portion of temperature specialists found in riparian seed banks indicates that even with available moisture riparian zone plant community composition will likely be impacted by changing temperatures. However, the presence of so many temperature generalists in the riparian zones suggests that some component of the seed bank is adapted to variable conditions and might offer resilience in a changing climate. Study results confirm the importance of conserving multiple hydrogeomorphic reach types because they support unique species assemblages.
ContributorsSetaro, Danika (Author) / Stromberg, Juliet (Thesis advisor) / Franklin, Janet (Committee member) / Makings, Elizabeth (Committee member) / Arizona State University (Publisher)
Created2016
Description
Adoptive transfer of T cells engineered to express synthetic antigen-specific T cell receptors (TCRs) has provocative therapeutic applications for treating cancer. However, expressing these synthetic TCRs in a CD4+ T cell line is a challenge. The CD4+ Jurkat T cell line expresses endogenous TCRs that compete for space, accessory proteins, and proliferative signaling, and there is the potential for mixed dimer formation between the α and β chains of the endogenous receptor and that of the synthetic cancer-specific TCRs. To prevent hybridization between the receptors and to ensure the binding affinity measured with flow cytometry analysis is between the tetramer and the TCR construct, a CRISPR-Cas9 gene editing pipeline was developed. The guide RNAs (gRNAs) within the complex were designed to target the constant region of the α and β chains, as they are conserved between TCR clonotypes. To minimize further interference and confer cytotoxic capabilities, gRNAs were designed to target the CD4 coreceptor, and the CD8 coreceptor was delivered in a mammalian expression vector. Further, Golden Gate cloning methods were validated in integrating the gRNAs into a CRISPR-compatible mammalian expression vector. These constructs were transfected via electroporation into CD4+ Jurkat T cells to create a CD8+ knockout TCR Jurkat cell line for broadly applicable uses in T cell immunotherapies.
ContributorsHirneise, Gabrielle Rachel (Author) / Anderson, Karen (Thesis advisor) / Mason, Hugh (Committee member) / Lake, Douglas (Committee member) / Arizona State University (Publisher)
Created2020