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Description
Flavivirus infections are emerging as significant threats to human health around the globe. Among them West Nile(WNV) and Dengue Virus (DV) are the most prevalent in causing human disease with WNV outbreaks occurring in all areas around the world and DV epidemics in more than 100 countries. WNV is a neurotropic virus capable of causing meningitis and encephalitis in humans. Currently, there are no therapeutic treatments or vaccines available. The expanding epidemic of WNV demands studies that develop efficacious therapeutics and vaccines and produce them rapidly and inexpensively. In response, our lab developed a plant-derived monoclonal antibody (mAb) (pHu-E16) against DIII (WNV antigen) that is able to neutralize and prevent mice from lethal infection. However, this drug has a short window of efficacy due to pHu-E16's inability to cross the Blood Brain Barrier (BBB) and enter the brain. Here, we constructed a bifunctional diabody, which couples the neutralizing activity of E16 and BBB penetrating activity of 8D3 mAb. We also produced a plant-derived E16 scFv-CH1-3 variant with equivalent specific binding as the full pHu-E16 mAb, but only requiring one gene construct for production. Furthermore, a WNV vaccine based on plant-derived DIII was developed showing proper folding and potentially protective immune response in mice. DV causes severe hemorrhaging diseases especially in people exposed to secondary DV infection from a heterotypic strain. It is hypothesized that sub-neutralizing cross-reactive antibodies from the first exposure aid the second infection in a process called antibody-dependent enhancement (ADE). ADE depends on the ability of mAb to bind Fc receptors (FcγRs), and has become a major roadblock for developing mAb-based therapeutics against DV. We aim to produce an anti-Dengue mAb (E60) in different glycoengineered plant lines that exhibit reduced/differential binding to FcγRs, therefore, reducing or eliminating ADE. We have successfully cloned the molecular constructs of E60, and expressed it in two plant lines with different glycosylation patterns. We demonstrated that both plant-derived E60 mAb glycoforms retained specific recognition and neutralization activity against DV. Overall, our study demonstrates great strives to develop efficacious therapeutics and potent vaccine candidates against Flaviviruses in plant expression systems.
ContributorsHurtado, Jonathan (Author) / Chen, Qiang (Thesis advisor) / Huffman, Holly A (Committee member) / Steele, Kelly P (Committee member) / Arizona State University (Publisher)
Created2014
Description
This dissertation consists of three substantive chapters. The first substantive chapter investigates the premature harvesting problem in fisheries. Traditionally, yield-per-recruit analysis has been used to both assess and address the premature harvesting of fish stocks. However, the fact that fish size often affects the unit price suggests that this approach may be inadequate. In this chapter, I first synthesize the conventional yield-per-recruit analysis, and then extend this conventional approach by incorporating a size-price function for a revenue-per-recruit analysis. An optimal control approach is then used to derive a general bioeconomic solution for the optimal harvesting of a short-lived single cohort. This approach prevents economically premature harvesting and provides an "optimal economic yield". By comparing the yield- and revenue-per-recruit management strategies with the bioeconomic management strategy, I am able to test the economic efficiency of the conventional yield-per-recruit approach. This is illustrated with a numerical study. It shows that a bioeconomic strategy can significantly improve economic welfare compared with the yield-per-recruit strategy, particularly in the face of high natural mortality. Nevertheless, I find that harvesting on a revenue-per-recruit basis improves management policy and can generate a rent that is close to that from bioeconomic analysis, in particular when the natural mortality is relatively low.
The second substantive chapter explores the conservation potential of a whale permit market under bounded economic uncertainty. Pro- and anti-whaling stakeholders are concerned about a recently proposed, "cap and trade" system for managing the global harvest of whales. Supporters argue that such an approach represents a novel solution to the current gridlock in international whale management. In addition to ethical objections, opponents worry that uncertainty about demand for whale-based products and the environmental benefits of conservation may make it difficult to predict the outcome of a whale share market. In this study, I use population and economic data for minke whales to examine the potential ecological consequences of the establishment of a whale permit market in Norway under bounded but significant economic uncertainty. A bioeconomic model is developed to evaluate the influence of economic uncertainties associated with pro- and anti- whaling demands on long-run steady state whale population size, harvest, and potential allocation. The results indicate that these economic uncertainties, in particular on the conservation demand side, play an important role in determining the steady state ecological outcome of a whale share market. A key finding is that while a whale share market has the potential to yield a wide range of allocations between conservation and whaling interests - outcomes in which conservationists effectively "buy out" the whaling industry seem most likely.
The third substantive chapter examines the sea lice externality between farmed fisheries and wild fisheries. A central issue in the debate over the effect of fish farming on the wild fisheries is the nature of sea lice population dynamics and the wild juvenile mortality rate induced by sea lice infection. This study develops a bioeconomic model that integrates sea lice population dynamics, fish population dynamics, aquaculture and wild capture salmon fisheries in an optimal control framework. It provides a tool to investigate sea lice control policy from the standpoint both of private aquaculture producers and wild fishery managers by considering the sea lice infection externality between farmed and wild fisheries. Numerical results suggest that the state trajectory paths may be quite different under different management regimes, but approach the same steady state. Although the difference in economic benefits is not significant in the particular case considered due to the low value of the wild fishery, I investigate the possibility of levying a tax on aquaculture production for correcting the sea lice externality generated by fish farms.
The second substantive chapter explores the conservation potential of a whale permit market under bounded economic uncertainty. Pro- and anti-whaling stakeholders are concerned about a recently proposed, "cap and trade" system for managing the global harvest of whales. Supporters argue that such an approach represents a novel solution to the current gridlock in international whale management. In addition to ethical objections, opponents worry that uncertainty about demand for whale-based products and the environmental benefits of conservation may make it difficult to predict the outcome of a whale share market. In this study, I use population and economic data for minke whales to examine the potential ecological consequences of the establishment of a whale permit market in Norway under bounded but significant economic uncertainty. A bioeconomic model is developed to evaluate the influence of economic uncertainties associated with pro- and anti- whaling demands on long-run steady state whale population size, harvest, and potential allocation. The results indicate that these economic uncertainties, in particular on the conservation demand side, play an important role in determining the steady state ecological outcome of a whale share market. A key finding is that while a whale share market has the potential to yield a wide range of allocations between conservation and whaling interests - outcomes in which conservationists effectively "buy out" the whaling industry seem most likely.
The third substantive chapter examines the sea lice externality between farmed fisheries and wild fisheries. A central issue in the debate over the effect of fish farming on the wild fisheries is the nature of sea lice population dynamics and the wild juvenile mortality rate induced by sea lice infection. This study develops a bioeconomic model that integrates sea lice population dynamics, fish population dynamics, aquaculture and wild capture salmon fisheries in an optimal control framework. It provides a tool to investigate sea lice control policy from the standpoint both of private aquaculture producers and wild fishery managers by considering the sea lice infection externality between farmed and wild fisheries. Numerical results suggest that the state trajectory paths may be quite different under different management regimes, but approach the same steady state. Although the difference in economic benefits is not significant in the particular case considered due to the low value of the wild fishery, I investigate the possibility of levying a tax on aquaculture production for correcting the sea lice externality generated by fish farms.
ContributorsHuang, Biao (Author) / Abbott, Joshua K (Thesis advisor) / Perrings, Charles (Thesis advisor) / Gerber, Leah R. (Committee member) / Muneepeerakul, Rachata (Committee member) / Schoon, Michael (Committee member) / Arizona State University (Publisher)
Created2014
Description
Immunotherapy has been revitalized with the advent of immune checkpoint blockade
treatments, and neo-antigens are the targets of immune system in cancer patients who
respond to the treatments. The cancer vaccine field is focused on using neo-antigens from
unique point mutations of genomic sequence in the cancer patient for making
personalized cancer vaccines. However, we choose a different path to find frameshift
neo-antigens at the mRNA level and develop broadly effective cancer vaccines based on
frameshift antigens.
In this dissertation, I have summarized and characterized all the potential frameshift
antigens from microsatellite regions in human, dog and mouse. A list of frameshift
antigens was validated by PCR in tumor samples and the mutation rate was calculated for
one candidate – SEC62. I develop a method to screen the antibody response against
frameshift antigens in human and dog cancer patients by using frameshift peptide arrays.
Frameshift antigens selected by positive antibody response in cancer patients or by MHC
predictions show protection in different mouse tumor models. A dog version of the
cancer vaccine based on frameshift antigens was developed and tested in a small safety
trial. The results demonstrate that the vaccine is safe and it can induce strong B and T cell
immune responses. Further, I built the human exon junction frameshift database which
includes all possible frameshift antigens from mis-splicing events in exon junctions, and I
develop a method to find potential frameshift antigens from large cancer
immunosignature dataset with these databases. In addition, I test the idea of ‘early cancer
diagnosis, early treatment’ in a transgenic mouse cancer model. The results show that
ii
early treatment gives significantly better protection than late treatment and the correct
time point for treatment is crucial to give the best clinical benefit. A model for early
treatment is developed with these results.
Frameshift neo-antigens from microsatellite regions and mis-splicing events are
abundant at mRNA level and they are better antigens than neo-antigens from point
mutations in the genomic sequences of cancer patients in terms of high immunogenicity,
low probability to cause autoimmune diseases and low cost to develop a broadly effective
vaccine. This dissertation demonstrates the feasibility of using frameshift antigens for
cancer vaccine development.
treatments, and neo-antigens are the targets of immune system in cancer patients who
respond to the treatments. The cancer vaccine field is focused on using neo-antigens from
unique point mutations of genomic sequence in the cancer patient for making
personalized cancer vaccines. However, we choose a different path to find frameshift
neo-antigens at the mRNA level and develop broadly effective cancer vaccines based on
frameshift antigens.
In this dissertation, I have summarized and characterized all the potential frameshift
antigens from microsatellite regions in human, dog and mouse. A list of frameshift
antigens was validated by PCR in tumor samples and the mutation rate was calculated for
one candidate – SEC62. I develop a method to screen the antibody response against
frameshift antigens in human and dog cancer patients by using frameshift peptide arrays.
Frameshift antigens selected by positive antibody response in cancer patients or by MHC
predictions show protection in different mouse tumor models. A dog version of the
cancer vaccine based on frameshift antigens was developed and tested in a small safety
trial. The results demonstrate that the vaccine is safe and it can induce strong B and T cell
immune responses. Further, I built the human exon junction frameshift database which
includes all possible frameshift antigens from mis-splicing events in exon junctions, and I
develop a method to find potential frameshift antigens from large cancer
immunosignature dataset with these databases. In addition, I test the idea of ‘early cancer
diagnosis, early treatment’ in a transgenic mouse cancer model. The results show that
ii
early treatment gives significantly better protection than late treatment and the correct
time point for treatment is crucial to give the best clinical benefit. A model for early
treatment is developed with these results.
Frameshift neo-antigens from microsatellite regions and mis-splicing events are
abundant at mRNA level and they are better antigens than neo-antigens from point
mutations in the genomic sequences of cancer patients in terms of high immunogenicity,
low probability to cause autoimmune diseases and low cost to develop a broadly effective
vaccine. This dissertation demonstrates the feasibility of using frameshift antigens for
cancer vaccine development.
ContributorsZhang, Jian (Author) / Johnston, Stephen Albert (Thesis advisor) / Chang, Yung (Committee member) / Stafford, Phillip (Committee member) / Chen, Qiang (Committee member) / Arizona State University (Publisher)
Created2018
Description
Prior to the COVID-19 global pandemic, ecotourism represented the tourism industry’s fastest growing segment with projections estimating that ecotourism would become the world’s largest tourism type by 2030. While the tourism industry will need several years to rebound, if historic trends tell us anything, it is that ecotourism will continue to represent a large portion of the overall industry and will continue to grow at a rate that outpaces all other tourism types. In theory, ecotourism promotes sustainable socioeconomic development while also minimizing negative environmental impacts. Unfortunately, research suggests that this is not always true, and many examples exist of ecotourism causing more harm than good. In order to combat these potential negative impacts, the ecotourism industry has become increasingly reliant on ecotourism certification programs to act as an assessment tool that identifies ecotourism’s best practitioners while minimizing false advertising present within the industry. Despite these beliefs in the efficacy of certification, there is a lack of empirical research to actually support certification as an effective assessment tool. Furthermore, little research has been conducted that assesses the impacts that certification itself has on ecotourism businesses (both certified and uncertified) and the local communities dependent on ecotourism.
My dissertation employs a mixed methods design and combines qualitative and quantitative research methods spanning multiple geographic scales to develop an understanding of certification programs as they exist today and to discern the impacts that certification itself may cause for all those either directly or indirectly involved in ecotourism. My findings ultimately suggest that certification reform is needed if certification programs are expected to be the assessment tool ecotourism experts claim them to be. Specifically, as certification exists presently, there is: no universal guideline or standard for existing certification programs to follow, a disconnect between the advertised benefits certification offers and the actual benefits received, and a lack of market penetration both amongst ecotourists and ecotourism businesses. Each of these must be addressed before certification can live up to its full potential. Furthermore, I found that certification may impact community socioeconomic dynamics, particularly by creating or exacerbating community wealth distribution.
My dissertation employs a mixed methods design and combines qualitative and quantitative research methods spanning multiple geographic scales to develop an understanding of certification programs as they exist today and to discern the impacts that certification itself may cause for all those either directly or indirectly involved in ecotourism. My findings ultimately suggest that certification reform is needed if certification programs are expected to be the assessment tool ecotourism experts claim them to be. Specifically, as certification exists presently, there is: no universal guideline or standard for existing certification programs to follow, a disconnect between the advertised benefits certification offers and the actual benefits received, and a lack of market penetration both amongst ecotourists and ecotourism businesses. Each of these must be addressed before certification can live up to its full potential. Furthermore, I found that certification may impact community socioeconomic dynamics, particularly by creating or exacerbating community wealth distribution.
ContributorsDavila, Ryan (Author) / Kinzig, Ann (Thesis advisor) / Perrings, Charles (Committee member) / Collins, James (Committee member) / Schoon, Michael (Committee member) / Buzinde, Christine (Committee member) / Arizona State University (Publisher)
Created2020
Description
Flaviviruses (FVs) are among the most medically important arboviruses of the world with the Dengue virus (DENV) accounting for a large percentage of infections observed in tropical and subtropical regions of the world. Globalization, travel, and the expanding range of mosquito vectors, such as Aedes aegypti, have increased the potential of infection rates and illnesses associated with FVs.
The DENV and the Zika (ZIKV) FVs frequently co-circulate and generally cause mild self-liming febrile illnesses. However, a secondary infection with a heterologous DENV serotype may lead to life threatening dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). DHF/DSS have been linked to antibody dependent enhancement of infection (ADE), a phenomenon that occurs when antibodies (Abs) formed against an initial infection with one serotype of DENV cross-reacts but does not neutralize a heterologous DENV serotype in a secondary infection. Furthermore, Abs raised against the ZIKV have been observed to cross-react with the DENV and vice versa, which can potentially cause ADE and lead to severe DENV disease. The ZIKV can be transmitted vertically and has been linked to devastating congenital defects such as microcephaly in newborns. FDA approved treatments do not exist for DENV and ZIKV illnesses. Thus, there is a need for safe and effective treatments for these co-circulating viruses. Here, a tetravalent bispecific antibody (bsAb) targeting the ZIKV and all four serotypes of the DENV was expressed in the Nicotiana benthamiana (N. benthamiana) plant. Functional assays of the DENV/ZIKV bsAb demonstrated binding, neutralization, and a significant reduction in ADE activity against both the DENV and the ZIKV.
A single chain variable fragment (scFv) and a diabody based on an antibody directed against the immune checkpoint inhibitor PD-L1, were also expressed in N. benthamiana leaves. The smaller sizes of the scFv and diabody confers them with the ability to penetrate deeper tissues making them beneficial in diagnostics, imaging, and possibly cancer therapy. The past few decades has seen long strives in recombinant protein production in plants with significant improvements in production, safety, and efficacy. These characteristics make plants an attractive platform for the production of recombinant proteins, biologics, and therapeutics.
The DENV and the Zika (ZIKV) FVs frequently co-circulate and generally cause mild self-liming febrile illnesses. However, a secondary infection with a heterologous DENV serotype may lead to life threatening dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). DHF/DSS have been linked to antibody dependent enhancement of infection (ADE), a phenomenon that occurs when antibodies (Abs) formed against an initial infection with one serotype of DENV cross-reacts but does not neutralize a heterologous DENV serotype in a secondary infection. Furthermore, Abs raised against the ZIKV have been observed to cross-react with the DENV and vice versa, which can potentially cause ADE and lead to severe DENV disease. The ZIKV can be transmitted vertically and has been linked to devastating congenital defects such as microcephaly in newborns. FDA approved treatments do not exist for DENV and ZIKV illnesses. Thus, there is a need for safe and effective treatments for these co-circulating viruses. Here, a tetravalent bispecific antibody (bsAb) targeting the ZIKV and all four serotypes of the DENV was expressed in the Nicotiana benthamiana (N. benthamiana) plant. Functional assays of the DENV/ZIKV bsAb demonstrated binding, neutralization, and a significant reduction in ADE activity against both the DENV and the ZIKV.
A single chain variable fragment (scFv) and a diabody based on an antibody directed against the immune checkpoint inhibitor PD-L1, were also expressed in N. benthamiana leaves. The smaller sizes of the scFv and diabody confers them with the ability to penetrate deeper tissues making them beneficial in diagnostics, imaging, and possibly cancer therapy. The past few decades has seen long strives in recombinant protein production in plants with significant improvements in production, safety, and efficacy. These characteristics make plants an attractive platform for the production of recombinant proteins, biologics, and therapeutics.
ContributorsEsqueda, Adrian (Author) / Chen, Qiang (Thesis advisor) / Arntzen, Charles (Committee member) / Lake, Douglas (Committee member) / Mason, Hugh (Committee member) / Arizona State University (Publisher)
Created2019
Description
Climate adaptation has not kept pace with climate impacts which has formed an adaptation gap. Increasingly insurance is viewed as a solution to close this gap. However, the efficacy and implications of using insurance in the climate adaptation space are not clear. Furthermore, past research has focused on specific actors or processes, not on the interactions and interconnections between the actors and the processes. I take a complex adaptive systems approach to map out how these dynamics are shaping adaptation and to interrogate what the insurance climate adaptation literature claims are the successes and pitfalls of insurance driving, enabling or being adaptation. From this interrogation it becomes apparent that insurance has enormous influence on its policy holders, builds telecoupling into local adaptation, and creates structures which support contradictory land use policies at the local level. Based on the influence insurance has on policy holders, I argue that insurance should be viewed as a form of governance. I synthesize insurance, governance and adaptation literature to examine exactly what governance tools insurance uses to exercise this influence and what the consequences may be. This research reveals that insurance may not be the exemplary adaptation approach the international community is hoping for. Using insurance, risk can be reduced without reducing vulnerability, and risk transfer can result in risk displacement which can reduce adaptation incentives, fuel maladaptation, or impose public burdens. Moreover, insurance requires certain information and legal relationships which can and often do structure that which is insured to the needs of insurance and shift authority away from governments to insurance companies or public-private partnerships. Each of these undermine the legitimacy of insurance-led local adaptation and contradict the stated social justice goals of international calls for insurance. Finally, I interrogate the potential justice concerns that emerged through an analysis of insurance as a form of adaptation governance. Using a multi-valent approach to justice I examine a suite of programs intended to support agricultural adaptation through insurance. This analysis demonstrates that although some programs clearly attempted to consider issues of justice, overall these existing programs raise distributional, procedural and recognition justice concerns.
ContributorsLueck, Vanessa (Author) / Klinsky, Sonja (Thesis advisor) / Schoon, Michael (Thesis advisor) / Eakin, Hallie (Committee member) / Arizona State University (Publisher)
Created2020
Description
Institutions (rules, norms, and shared strategies) are social feedback systems that structure actors’ decision-making context. It is important to investigate institutional design to understand how rules interact and generate feedbacks that affect robustness, i.e., the ability to respond to change. This is particularly important when assessing sustainable use/conservation trade-offs that affect species’ long-term survival. My research utilized the institutional grammar (IG) and robust institutional design to investigate these linkages in the context of four international conservation treaties.
First, the IG was used to code the regulatory formal treaty rules. The coded statements were then assessed to determine the rule linkages and dynamic interactions with a focus on monitoring and related reporting and enforcement mechanisms. Treaties with a regulatory structure included a greater number and more tightly linked rules related to these mechanisms than less regulatory instruments. A higher number of actors involved in these activities at multiple levels also seemed critical to a well-functioning monitoring system.
Then, drawing on existing research, I built a set of constitutive rule typologies to supplement the IG and code the treaties’ constitutive rules. I determined the level of fit between the constitutive and regulatory rules by examining the monitoring mechanisms, as well as treaty opt-out processes. Treaties that relied on constitutive rules to guide actor decision-making generally exhibited gaps and poorer rule fit. Regimes which used constitutive rules to provide actors with information related to the aims, values, and context under which regulatory rules were being advanced tended to exhibit better fit, rule consistency, and completeness.
The information generated in the prior studies, as well as expert interviews, and the analytical frameworks of Ostrom’s design principles, fit, and polycentricity, then aided the analysis of treaty robustness. While all four treaties were polycentric, regulatory regimes exhibited strong information processing feedbacks as evidenced by the presence of all design principles (in form and as perceived by experts) making them theoretically more robust to change than non-regulatory ones. Interestingly, treaties with contested decision-making seemed more robust to change indicating contestation facilitates robust decision-making or its effects are ameliorated by rule design.
First, the IG was used to code the regulatory formal treaty rules. The coded statements were then assessed to determine the rule linkages and dynamic interactions with a focus on monitoring and related reporting and enforcement mechanisms. Treaties with a regulatory structure included a greater number and more tightly linked rules related to these mechanisms than less regulatory instruments. A higher number of actors involved in these activities at multiple levels also seemed critical to a well-functioning monitoring system.
Then, drawing on existing research, I built a set of constitutive rule typologies to supplement the IG and code the treaties’ constitutive rules. I determined the level of fit between the constitutive and regulatory rules by examining the monitoring mechanisms, as well as treaty opt-out processes. Treaties that relied on constitutive rules to guide actor decision-making generally exhibited gaps and poorer rule fit. Regimes which used constitutive rules to provide actors with information related to the aims, values, and context under which regulatory rules were being advanced tended to exhibit better fit, rule consistency, and completeness.
The information generated in the prior studies, as well as expert interviews, and the analytical frameworks of Ostrom’s design principles, fit, and polycentricity, then aided the analysis of treaty robustness. While all four treaties were polycentric, regulatory regimes exhibited strong information processing feedbacks as evidenced by the presence of all design principles (in form and as perceived by experts) making them theoretically more robust to change than non-regulatory ones. Interestingly, treaties with contested decision-making seemed more robust to change indicating contestation facilitates robust decision-making or its effects are ameliorated by rule design.
ContributorsBrady, Ute (Author) / Anderies, John M. (Thesis advisor) / Schoon, Michael (Committee member) / Minteer, Ben A. (Committee member) / Gerber, Leah (Committee member) / Siddiki, Saba (Committee member) / Arizona State University (Publisher)
Created2020
Description
This dissertation focuses on water security in terms of sustaining socio-economic development, livelihoods, and human well-being. Using the double exposure framework, I analyze the combined effect of climate change and economic development on water security in the Philippines. There is a need to examine how the combination of these two processes aggravate existing inequalities related to water security among different groups of people, and also analyze how these two processes can combine to increase stakeholders’ vulnerability to water-related shocks and stresses. The Philippines has been rated as one of the countries that is most vulnerable to climate change due to its exposure to extreme climate events and sea level rise. At the same time, the Philippines is currently undergoing an economic transition from a predominantly agricultural country to one where industry and services play a larger role. This dissertation zeroes in on the water security of municipalities in the Philippines, which were sorted into different syndromes based on a combination of their risk to future hydro-climatic changes and economic growth trends. Four syndromes which covered 73% of the population then emerged. By comparing five case study municipalities drawn from these four syndromes, I offer insights into how different combinations of climatic and economic factors can impact water security, and which combination could have the lowest water security in the future. Through analyzing the results of focus group discussions and semi-structured interviews, I also explore the variation of perceptions and collaborative strategies of stakeholders regarding their current and future water security. While each municipality had different climate and economic vulnerabilities, they shared largely similar water security perceptions and used the same strategies.
ContributorsLorenzo, Theresa Marie (Author) / Kinzig, Ann (Thesis advisor) / David, Carlos Primo (Committee member) / Perrings, Charles (Committee member) / Schoon, Michael (Committee member) / Selin, Cynthia (Committee member) / Arizona State University (Publisher)
Created2021
Description
Influenza is a deadly disease that poses a major threat to global health. The surface proteins of influenza A, the type most often associated with epidemics and pandemics, mutate at a very high frequency from season to season, reducing the efficacy of seasonal influenza vaccines. However, certain regions of these proteins are conserved between strains of influenza A, making them attractive targets for the development of a ‘universal’ influenza vaccine. One of these highly conserved regions is the ectodomain of the influenza matrix 2 protein (M2e). Studies have shown that M2e is poorly immunogenic on its own, but when properly adjuvanted it can be used to induce protective immune responses against many strains of influenza A. In this thesis, M2e was fused to a pair experimental ‘vaccine platforms’: an antibody fusion protein designed to assemble into a recombinant immune complex (RIC) and the hepatitis B core antigen (HBc) that can assemble into virus-like particles (VLP). The two antigens were produced in Nicotiana benthamiana plants through the use of geminiviral vectors and were subsequently evaluated in mouse trials. Mice were administered three doses of either the VLP alone or a 1:1 combination of the VLP and the RIC, and recipients of both the VLP and RIC exhibited endpoint anti-M2e antibody titers that were 2 to 3 times higher than mice that received the VLP alone. While IgG2a:IgG1 ratios, which can suggest the type of immune response (TH1 vs TH2) an antigen will elicit, were higher in mice vaccinated solely with the VLP, the higher overall titers are encouraging and demonstrate a degree of interaction between the RIC and VLP vaccines. Further research is necessary to determine the optimal balance of VLP and RIC to maximize IgG2a:IGg1 ratios as well as whether such interaction would be observed through the use of a variety of diverse antigens, though the results of other studies conducted in this lab suggests that this is indeed the case. The results of this study demonstrate not only the successful development of a promising new universal influenza A vaccine, but also that co-delivering different types of recombinant vaccines could reduce the total number of vaccine doses needed to achieve a protective immune response.
ContributorsFavre, Brandon Chetan (Author) / Mason, Hugh S (Thesis advisor) / Mor, Tsafrir (Committee member) / Chen, Qiang (Committee member) / Arizona State University (Publisher)
Created2019
Description
Environmental stressors can perturb cellular homeostasis. Cells activate an integrated stress response that will alleviate the effects of the ongoing stress. Stress-activated protein kinases function to phosphorylate the eukaryotic translation initiation factor, eIF2α, which results in inhibition of translation of house-keeping genes. Following these events, formation of cytoplasmic messenger ribonucleoprotein complexes, known as stress granules, will take place. Stress granules typically have a pro-survival function. These studies demonstrate that assembly of stress granules can also lead to necroptosis. Necroptosis is a caspase-independent, receptor-interacting protein kinase 3 (RIPK3)-dependent cell death pathway executed by mixed lineage kinase domain-like (MLKL) protein. Cellular stress is induced using arsenite (oxidative stress) or by infection with vaccinia virus (VACV) E3 protein Z-DNA-binding domain mutant, VACV-E3LΔ83N. In both cases, RIPK3-dependent death was observed in interferon (IFN)-primed L929 cells. This death led to phosphorylation and trimerization of MLKL, indicative of necroptosis. Necroptosis induced by oxidative stress and VACV-E3LΔ83N infection was dependent on the host Z-form nucleic acid sensor, DNA-dependent activator of IFN-regulatory factors (DAI), as it was inhibited in DAI-deficient L929 cells. Under both cellular stresses, DAI associated with RIPK3 and formed high-molecular-weight complexes, consistent with formation of the necrosomes. DAI localized into stress granules during necroptosis induced by arsenite and the mutant virus, and the necrosomes formed only in presence of stress granule assembly. The significance of stress granules for cellular stress-induced necroptosis was demonstrated using knock-out (KO) cell lines unable to form granules: T cell-restricted intracellular antigen 1 (TIA-1) KO MEF cells and Ras GTPase-activating protein-binding proteins 1 and 2 (G3BP1/2) KO U2OS cells. Necroptosis was inhibited in absence of stress granule formation as no cell death or activation of MLKL was observed in the knock-out cell lines following arsenite treatment or VACV-E3LΔ83N infection. Furthermore, wild-type VACV was able to inhibit stress granule assembly, which coincided with the virus ability to inhibit necroptosis. These studies have led to a model of Z-form nucleic acids being involved in activation of the stress granule-mediated necroptosis following induction by environmental stressors. These results have significance for understanding the etiology of human diseases and the antiviral innate immunity.
ContributorsSzczerba, Mateusz Bartlomiej (Author) / Jacobs, Bertram L (Thesis advisor) / Langland, Jeffrey (Committee member) / Lake, Douglas (Committee member) / Chen, Qiang (Committee member) / Arizona State University (Publisher)
Created2021