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
Two nearly homogenous 60 acre watersheds near Heber, Arizona, within the Apache-Sitgreaves National Forest, were burned at moderate and high severities during the 2002 Rodeo-Chediski wildfire. Each watershed had 30 permanent plots located on it from earlier studies. In 2011, nearly 10 years following the fire, the plots were re-measured to determine how fire severity affects the long term vegetative recovery of this ecosystem; specifically herbaceous production and tree regeneration and density. Canopy cover, litter depth, herbaceous weight, herbaceous cover and shrub cover are vital indicators of herbaceous production, and were found to be significantly different between the sites. Canopy cover and litter depth were found to be significantly higher on the moderate site while herbaceous weight, herbaceous cover and shrub cover were found to be significantly higher on the high site. Tree densities of the three present tree species, ponderosa pine, alligator juniper, and gambel oak, were measured and divided into five size classes to distinguish the diversity of the communities. The mean densities for each species and size class were analyzed to determine if there were any statistically significant differences between the sites. Ponderosa pine saplings (regeneration) were found to have no significant differences between the sites. Juniper and oak saplings were found to be significantly higher on the high site. The remaining four ponderosa pine size classes were found to be significantly higher on the moderate site while the remaining four size classes for juniper and oak were found to have no statistical differences between the sites. Further analysis of the tree proportions revealed that the ponderosa pine species was significantly higher on the moderate site while juniper and oak were significantly higher on the high site. Species specific proportion analysis showed that the ponderosa pine size classes were significantly different across the sites while the juniper and oak size classes showed no significant differences between the sites. Within the ponderosa pine size classes, saplings were found to be significantly higher on the high site while the remaining four classes were significantly higher on the moderate site.
ContributorsNeeley, Heidi L (Author) / Alford, Eddie (Thesis advisor) / Pyne, Stephen (Committee member) / Brady, Ward (Committee member) / Arizona State University (Publisher)
Created2012
Description
ABSTRACT The elephant tree, Bursera microphylla, is at the northern limit of its range in central Arizona. This species is sensitive to frost damage thus limiting its occurrence in more northern areas of the southwest. Marginal populations of B. microphylla are found in mountain ranges of Central Arizona and are known to occur in the rugged mountain range system of the South Mountain Municipal Park (SMMP). Little is known of the distribution of this species within the park and details relevant to the health of both individual plants and the population such as diameter and number of trunks, height, and presence of damage have not been examined. This study was designed, in part, to test the hypothesis that favorable microhabitats at SMMP are created by particular combinations of abiotic features including aspect, slope, elevation and solar radiation. Data on abiotic factors, as well as specific individual plant locations and characteristics were obtained for 100 individuals. Temperature data was collected in vertical transects at different altitudinal levels. Some of these data were used in spatial analyses to generate a habitat suitability model using GIS software. Furthermore, collected data was analyzed using Matlab© software to identify potential trends in the variation of morphological traits. In addition, for comparative purposes similar information at one hundred computer-generated randomly chosen points throughout SMMP was obtained. The GIS spatial analyses indicated that aspect, slope, elevation, and relative solar radiance are strongly associated as major climatic components of the microhabitat of B. microphylla. Temperature data demonstrated that there are significant differences in ambient temperature among different altitudinal gradients with middle elevations being more favorable. Furthermore, analyses performed using Matlab© to explore trends of elevation as a factor indicated that multiple trunk plants are more commonly found at higher elevations than single trunk plants, there is a positive correlation of trunk diameter with elevation, and that canopy volume has a negative correlation with respect to elevation. It was concluded that microhabitats where B. microphylla occurs at the northern limit of its range require a particular combination of abiotic features that can be easily altered by climatic changes.
ContributorsCordova, Cesar, M.S (Author) / Steele, Kelly P. (Thesis advisor) / Tridane, Abdessaman (Committee member) / Miller, William (Committee member) / Brady, Ward (Committee member) / Arizona State University (Publisher)
Created2011
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