Matching Items (22)
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
This paper presents a multiscale modeling approach to simulating the self-sensing behavior of a load sensitive smart polymer material. A statistical spring-bead based network model is developed to bridge the molecular dynamics simulations at the nanoscale and the finite element model at the macroscale. Parametric studies are conducted on the developed network model to investigate the effects of the thermoset crosslinking degree on the mechanical response of the self-sensing material. A comparison between experimental and simulation results shows that the multiscale framework is able to capture the global mechanical response with adequate accuracy and the network model is also capable of simulating the self-sensing phenomenon of the smart polymer. Finally, the molecular dynamics simulation and network model based simulation are implemented to evaluate damage initiation in the self-sensing material under monotonic loading.
ContributorsZhang, Jinjun (Author) / Koo, Bonsung (Author) / Liu, Yingtao (Author) / Zou, Jin (Author) / Chattopadhyay, Aditi (Author) / Dai, Lenore (Author) / Ira A. Fulton Schools of Engineering (Contributor) / School for the Engineering of Matter, Transport and Energy (Contributor)
Created2015-08-01
Description
Wall-bounded turbulence manifests itself in a broad range of applications, not least of which in hydraulic systems. Here we briefly review the significant advances over the past few decades in the fundamental study of wall turbulence over smooth and rough surfaces, with an emphasis on coherent structures and their role at high Reynolds numbers. We attempt to relate these findings to parallel efforts in the hydraulic engineering community and discuss the implications of coherent structures in important hydraulic phenomena.
ContributorsAdrian, Ronald (Author) / Marusic, Ivan (Author) / Ira A. Fulton Schools of Engineering (Contributor) / School for the Engineering of Matter, Transport and Energy (Contributor)
Created2012-09-10
Description
Vortex organization in the outer layer of a turbulent boundary layer overlying sparse, hemispherical roughness elements is explored with two-component particle-image velocimetry (PIV) in multiple streamwise-wall-normal measurement planes downstream and between elements. The presence of sparse roughness elements causes a shortening of the streamwise length scale in the near-wall region. These measurements confirm that vortex packets exist in the outer layer of flow over rough walls, but that their organization is altered, and this is interpreted as the underlying cause of the length-scale reduction. In particular, the elements shed vortices which appear to align in the near-wall region, but are distinct from the packets. Further, it is observed that ejection events triggered in the element wakes are more intense compared to the ejection events in smooth wall. We speculate that this may initiate a self-sustaining mechanism leading to the formation of hairpin packets as a much more effective instability compared to those typical of smooth-wall turbulence.
ContributorsGuala, Michael (Author) / Tomkins, Christopher D. (Author) / Christensen, Kenneth T. (Author) / Adrian, Ronald (Author) / Ira A. Fulton Schools of Engineering (Contributor) / School for the Engineering of Matter, Transport and Energy (Contributor)
Created2012-09-09
Description
The dynamic importance of spanwise vorticity and vortex filaments has been assessed in steady, uniform open-channel flows by means of particle image velocimetry (PIV). By expressing the net force due to Reynolds’ turbulent shear stress, ∂(−[bar over uv]) ∂y, in terms of two velocity-vorticity correlations, [bar over vω[subscript z]] and [bar over wω[subscript y]], the results show that both spanwise vorticity [bar over ω[subscript z]] and the portion of it that is due to spanwise filaments make important contributions to the net force and hence the shape of the mean flow profile. Using the swirling strength to identify spanwise vortex filaments, it is found that they account for about 45% of [bar over vω[subscript z]], the remainder coming from non-filamentary spanwise vorticity, i.e. shear. The mechanism underlying this contribution is the movement of vortex filaments away from the wall. The contribution of spanwise vortex filaments to the Reynolds stress is small because they occupy a small fraction of the flow. The contribution of the induced motion of the spanwise vortex filaments is significant.
ContributorsChen, Qigang (Author) / Adrian, Ronald (Author) / Zhong, Qiang (Author) / Li, Danxun (Author) / Wang, Xingkui (Author) / Ira A. Fulton Schools of Engineering (Contributor) / School for the Engineering of Matter, Transport and Energy (Contributor)
Created2013-11-30
Description
X-ray free-electron lasers provide novel opportunities to conduct single particle analysis on nanoscale particles. Coherent diffractive imaging experiments were performed at the Linac Coherent Light Source (LCLS), SLAC National Laboratory, exposing single inorganic core-shell nanoparticles to femtosecond hard-X-ray pulses. Each facetted nanoparticle consisted of a crystalline gold core and a differently shaped palladium shell. Scattered intensities were observed up to about 7 nm resolution. Analysis of the scattering patterns revealed the size distribution of the samples, which is consistent with that obtained from direct real-space imaging by electron microscopy. Scattering patterns resulting from single particles were selected and compiled into a dataset which can be valuable for algorithm developments in single particle scattering research.
ContributorsLi, Xuanxuan (Author) / Chiu, Chun-Ya (Author) / Wang, Hsiang-Ju (Author) / Kassemeyer, Stephan (Author) / Botha, Sabine (Author) / Shoeman, Robert L. (Author) / Lawrence, Robert (Author) / Kupitz, Christopher (Author) / Kirian, Richard (Author) / James, Daniel (Author) / Wang, Dingjie (Author) / Nelson, Garrett (Author) / Messerschmidt, Marc (Author) / Boutet, Sebastien (Author) / Williams, Garth J. (Author) / Hartman, Elisabeth (Author) / Jafarpour, Aliakbar (Author) / Foucar, Lutz M. (Author) / Barty, Anton (Author) / Chapman, Henry (Author) / Liang, Mengning (Author) / Menzel, Andreas (Author) / Wang, Fenglin (Author) / Basu, Shibom (Author) / Fromme, Raimund (Author) / Doak, R. Bruce (Author) / Fromme, Petra (Author) / Weierstall, Uwe (Author) / Huang, Michael H. (Author) / Spence, John (Author) / Schlichting, Ilme (Author) / Hogue, Brenda (Author) / Liu, Haiguang (Author) / ASU Biodesign Center Immunotherapy, Vaccines and Virotherapy (Contributor) / Biodesign Institute (Contributor) / Applied Structural Discovery (Contributor) / College of Liberal Arts and Sciences (Contributor) / School of Molecular Sciences (Contributor) / Department of Physics (Contributor) / School of Life Sciences (Contributor)
Created2017-04-11
Description
It is widely anticipated that a prophylactic vaccine may be needed to control the HIV/AIDS epidemic worldwide. Despite over two decades of research, a vaccine against HIV-1 remains elusive, although a recent clinical trial has shown promising results. Recent studies have focused on highly conserved domains within HIV-1 such as the membrane proximal external region (MPER) of the envelope glycoprotein, gp41. MPER has been shown to play critical roles in mucosal transmission of HIV-1, though this peptide is poorly immunogenic on its own. Here we provide evidence that plant-produced HIV-1 enveloped virus-like particles (VLPs) consisting of Gag and a deconstructed form of gp41 comprising the MPER, transmembrane, and cytoplasmic domains (Dgp41) provides an effective platform to display MPER for use as an HIV vaccine candidate. 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 investigated in BALB/c mice. Serum antibody responses against both the Gag and gp41 antigens were elicited when systemically primed with VLPs. These responses could be recalled following systemic boosting with VLPs. In addition, mucosal priming with VLPs allowed for a boosting response against Gag and gp41 when boosted with either candidate. Importantly, the VLPs also induced Gag-specific CD4 and CD8 T-cell responses. This report on the immunogenicity of plant-based Gag/Dgp41 VLPs may represent an important milestone on the road towards a broadly efficacious and inexpensive subunit vaccine against HIV-1.
ContributorsKessans, Sarah (Author) / Linhart, Mark (Author) / Meador, Lydia (Author) / Kilbourne, Jacquelyn (Author) / Hogue, Brenda (Author) / Fromme, Petra (Author) / Matoba, Nobuyuki (Author) / Mor, Tsafrir (Author) / College of Liberal Arts and Sciences (Contributor) / School of Life Sciences (Contributor) / Biodesign Institute (Contributor, Contributor) / Infectious Diseases and Vaccinology (Contributor) / Applied Structural Discovery (Contributor) / Department of Chemistry and Biochemistry (Contributor)
Created2016-03-17
Description
Diacylglycerol kinase catalyses the ATP-dependent conversion of diacylglycerol to phosphatidic acid in the plasma membrane of Escherichia coli. The small size of this integral membrane trimer, which has 121 residues per subunit, means that available protein must be used economically to craft three catalytic and substrate-binding sites centred about the membrane/cytosol interface. How nature has accomplished this extraordinary feat is revealed here in a crystal structure of the kinase captured as a ternary complex with bound lipid substrate and an ATP analogue. Residues, identified as essential for activity by mutagenesis, decorate the active site and are rationalized by the ternary structure. The γ-phosphate of the ATP analogue is positioned for direct transfer to the primary hydroxyl of the lipid whose acyl chain is in the membrane. A catalytic mechanism for this unique enzyme is proposed. The active site architecture shows clear evidence of having arisen by convergent evolution.
ContributorsLi, Dianfan (Author) / Stansfeld, Phillip J. (Author) / Sansom, Mark S. P. (Author) / Keogh, Aaron (Author) / Vogeley, Lutz (Author) / Howe, Nicole (Author) / Lyons, Joseph A. (Author) / Aragao, David (Author) / Fromme, Petra (Author) / Fromme, Raimund (Author) / Basu, Shibom (Author) / Grotjohann, Ingo (Author) / Kupitz, Christopher (Author) / Rendek, Kimberley (Author) / Weierstall, Uwe (Author) / Zatsepin, Nadia (Author) / Cherezov, Vadim (Author) / Liu, Wei (Author) / Bandaru, Sateesh (Author) / English, Niall J. (Author) / Gati, Cornelius (Author) / Barty, Anton (Author) / Yefanov, Oleksandr (Author) / Chapman, Henry N. (Author) / Diederichs, Kay (Author) / Messerschmidt, Marc (Author) / Boutet, Sebastien (Author) / Williams, Garth J. (Author) / Seibert, M. Marvin (Author) / Caffrey, Martin (Author) / College of Liberal Arts and Sciences (Contributor) / School of Molecular Sciences (Contributor) / Biodesign Institute (Contributor) / Applied Structural Discovery (Contributor) / Department of Physics (Contributor)
Created2015-12-17
Description
Phytochromes are a family of photoreceptors that control light responses of plants, fungi and bacteria. A sequence of structural changes, which is not yet fully understood, leads to activation of an output domain. Time-resolved serial femtosecond crystallography (SFX) can potentially shine light on these conformational changes. Here we report the room temperature crystal structure of the chromophore-binding domains of the Deinococcus radiodurans phytochrome at 2.1 Å resolution. The structure was obtained by serial femtosecond X-ray crystallography from microcrystals at an X-ray free electron laser. We find overall good agreement compared to a crystal structure at 1.35 Å resolution derived from conventional crystallography at cryogenic temperatures, which we also report here. The thioether linkage between chromophore and protein is subject to positional ambiguity at the synchrotron, but is fully resolved with SFX. The study paves the way for time-resolved structural investigations of the phytochrome photocycle with time-resolved SFX.
ContributorsEdlund, Petra (Author) / Takala, Heikki (Author) / Claesson, Elin (Author) / Henry, Leocadie (Author) / Dods, Robert (Author) / Lehtivuori, Heli (Author) / Panman, Matthijs (Author) / Pande, Kanupriya (Author) / White, Thomas (Author) / Nakane, Takanori (Author) / Berntsson, Oskar (Author) / Gustavsson, Emil (Author) / Bath, Petra (Author) / Modi, Vaibhav (Author) / Roy Chowdhury, Shatabdi (Author) / Zook, James (Author) / Berntsen, Peter (Author) / Pandey, Suraj (Author) / Poudyal, Ishwor (Author) / Tenboer, Jason (Author) / Kupitz, Christopher (Author) / Barty, Anton (Author) / Fromme, Petra (Author) / Koralek, Jake D. (Author) / Tanaka, Tomoyuki (Author) / Spence, John (Author) / Liang, Mengning (Author) / Hunter, Mark S. (Author) / Boutet, Sebastien (Author) / Nango, Eriko (Author) / Moffat, Keith (Author) / Groenhof, Gerrit (Author) / Ihalainen, Janne (Author) / Stojkovic, Emina A. (Author) / Schmidt, Marius (Author) / Westenhoff, Sebastian (Author) / College of Liberal Arts and Sciences (Contributor) / School of Molecular Sciences (Contributor) / Biodesign Institute (Contributor) / Applied Structural Discovery (Contributor) / Department of Physics (Contributor)
Created2016-10-19
Description
Because metallic aircraft components are subject to a variety of in-service loading conditions, predicting their fatigue life has become a critical challenge. To address the failure mode mitigation of aircraft components and at the same time reduce the life-cycle costs of aerospace systems, a reliable prognostics framework is essential. In this paper, a hybrid prognosis model that accurately predicts the crack growth regime and the residual-useful-life estimate of aluminum components is developed. The methodology integrates physics-based modeling with a data-driven approach. Different types of loading conditions such as constant amplitude, random, and overload are investigated. The developed methodology is validated on an Al 2024-T351 lug joint under fatigue loading conditions. The results indicate that fusing the measured data and physics-based models improves the accuracy of prediction compared to a purely data-driven or physics-based approach.
ContributorsNeerukatti, Rajesh Kumar (Author) / Liu, Kuang C. (Author) / Kovvali, Narayan (Author) / Chattopadhyay, Aditi (Author) / Ira A. Fulton Schools of Engineering (Contributor) / School for the Engineering of Matter, Transport and Energy (Contributor)
Created2014-04-01