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Syrian hamsters infected with Andes virus (ANDV) develop a disease that recapitulates many of the salient features of human hantavirus pulmonary syndrome (HPS), including lethality. Infection of hamsters with Hantaan virus (HTNV) results in an asymptomatic, disseminated infection. In order to explore this dichotomy, we examined the transcriptome of ANDV- and HTNV-infected hamsters. Using NanoString technology, we examined kinetic transcriptional responses in whole blood collected from ANDV- and HTNV-infected hamsters. Of the 770 genes analyzed, key differences were noted in the kinetics of type I interferon sensing and signaling responses, complement activation, and apoptosis pathways between ANDV- and HTNV-infected hamsters. Delayed activation of type I interferon responses in ANDV-infected hamsters represents a potential mechanism that ANDV uses to subvert host immune responses and enhance disease. This is the first genome-wide analysis of hantavirus-infected hamsters and provides insight into potential avenues for therapeutics to hantavirus disease.
Rebecca L. Brocato; Louis A. Altamura; Brian D. Carey; Casey C. Perley; Candace D. Blancett; Timothy D. Minogue; Jay W. Hooper. Comparison of transcriptional responses between pathogenic and nonpathogenic hantavirus infections in Syrian hamsters using NanoString. PLOS Neglected Tropical Diseases 2021, 15, 1 .
AMA StyleRebecca L. Brocato, Louis A. Altamura, Brian D. Carey, Casey C. Perley, Candace D. Blancett, Timothy D. Minogue, Jay W. Hooper. Comparison of transcriptional responses between pathogenic and nonpathogenic hantavirus infections in Syrian hamsters using NanoString. PLOS Neglected Tropical Diseases. 2021; 15 (8):1.
Chicago/Turabian StyleRebecca L. Brocato; Louis A. Altamura; Brian D. Carey; Casey C. Perley; Candace D. Blancett; Timothy D. Minogue; Jay W. Hooper. 2021. "Comparison of transcriptional responses between pathogenic and nonpathogenic hantavirus infections in Syrian hamsters using NanoString." PLOS Neglected Tropical Diseases 15, no. 8: 1.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a highly transmissible RNA virus that is the causative agent of the Coronavirus disease 2019 (COVID-19) pandemic. Patients with severe COVID-19 may develop acute lung injury (ALI) or acute respiratory distress syndrome (ARDS) and require mechanical ventilation. Key features of SARS-CoV-2 induced pulmonary complications include an overexpression of pro-inflammatory chemokines and cytokines that contribute to a ‘cytokine storm.’ In the current study an inflammatory state in Calu-3 human lung epithelial cells was characterized in which significantly elevated transcripts of the immunostimulatory chemokines CXCL9, CXCL10, and CXCL11 were present. Additionally, an increase in gene expression of the cytokines IL-6, TNFα, and IFN-γ was observed. The transcription of CXCL9, CXCL10, IL-6, and IFN-γ was also induced in the lungs of human transgenic angiotensin converting enzyme 2 (ACE2) mice infected with SARS-CoV-2. To elucidate cell signaling pathways responsible for chemokine upregulation in SARS-CoV-2 infected cells, small molecule inhibitors targeting key signaling kinases were used. The induction of CXCL9, CXCL10, and CXCL11 gene expression in response to SARS-CoV-2 infection was markedly reduced by treatment with the AKT inhibitor GSK690693. Samples from COVID-19 positive individuals also displayed marked increases in CXCL9, CXCL10, and CXCL11 transcripts as well as transcripts in the AKT pathway. The current study elucidates potential pathway specific targets for reducing the induction of chemokines that may be contributing to SARS-CoV-2 pathogenesis via hyperinflammation.
Victoria Callahan; Seth Hawks; Matthew Crawford; Caitlin Lehman; Holly Morrison; Hannah Ivester; Ivan Akhrymuk; Niloufar Boghdeh; Rafaela Flor; Carla Finkielstein; Irving Allen; James Weger-Lucarelli; Nisha Duggal; Molly Hughes; Kylene Kehn-Hall. The Pro-Inflammatory Chemokines CXCL9, CXCL10 and CXCL11 Are Upregulated Following SARS-CoV-2 Infection in an AKT-Dependent Manner. Viruses 2021, 13, 1062 .
AMA StyleVictoria Callahan, Seth Hawks, Matthew Crawford, Caitlin Lehman, Holly Morrison, Hannah Ivester, Ivan Akhrymuk, Niloufar Boghdeh, Rafaela Flor, Carla Finkielstein, Irving Allen, James Weger-Lucarelli, Nisha Duggal, Molly Hughes, Kylene Kehn-Hall. The Pro-Inflammatory Chemokines CXCL9, CXCL10 and CXCL11 Are Upregulated Following SARS-CoV-2 Infection in an AKT-Dependent Manner. Viruses. 2021; 13 (6):1062.
Chicago/Turabian StyleVictoria Callahan; Seth Hawks; Matthew Crawford; Caitlin Lehman; Holly Morrison; Hannah Ivester; Ivan Akhrymuk; Niloufar Boghdeh; Rafaela Flor; Carla Finkielstein; Irving Allen; James Weger-Lucarelli; Nisha Duggal; Molly Hughes; Kylene Kehn-Hall. 2021. "The Pro-Inflammatory Chemokines CXCL9, CXCL10 and CXCL11 Are Upregulated Following SARS-CoV-2 Infection in an AKT-Dependent Manner." Viruses 13, no. 6: 1062.
Venezuelan equine encephalitis virus (VEEV) is an alphavirus that causes encephalitis. Previous work indicated that VEEV infection induced early growth response 1 (EGR1) expression, leading to cell death via the protein kinase R (PKR)-like endoplasmic reticulum kinase (PERK) arm of the unfolded protein response (UPR) pathway. Loss of PERK prevented EGR1 induction and decreased VEEV-induced death. The results presented within show that loss of PERK in human primary astrocytes dramatically reduced VEEV and eastern equine encephalitis virus (EEEV) infectious titers by 4–5 log10. Loss of PERK also suppressed VEEV replication in primary human pericytes and human umbilical vein endothelial cells, but it had no impact on VEEV replication in transformed U87MG and 293T cells. A significant reduction in VEEV RNA levels was observed as early as 3 h post-infection, but viral entry assays indicated that the loss of PERK minimally impacted VEEV entry. In contrast, the loss of PERK resulted in a dramatic reduction in viral nonstructural protein translation and negative-strand viral RNA production. The loss of PERK also reduced the production of Rift Valley fever virus and Zika virus infectious titers. These data indicate that PERK is an essential factor for the translation of alphavirus nonstructural proteins and impacts multiple RNA viruses, making it an exciting target for antiviral development.
Bibha Dahal; Caitlin Lehman; Ivan Akhrymuk; Nicole Bracci; Lauren Panny; Michael Barrera; Nishank Bhalla; Jonathan Jacobs; Jonathan Dinman; Kylene Kehn-Hall. PERK Is Critical for Alphavirus Nonstructural Protein Translation. Viruses 2021, 13, 892 .
AMA StyleBibha Dahal, Caitlin Lehman, Ivan Akhrymuk, Nicole Bracci, Lauren Panny, Michael Barrera, Nishank Bhalla, Jonathan Jacobs, Jonathan Dinman, Kylene Kehn-Hall. PERK Is Critical for Alphavirus Nonstructural Protein Translation. Viruses. 2021; 13 (5):892.
Chicago/Turabian StyleBibha Dahal; Caitlin Lehman; Ivan Akhrymuk; Nicole Bracci; Lauren Panny; Michael Barrera; Nishank Bhalla; Jonathan Jacobs; Jonathan Dinman; Kylene Kehn-Hall. 2021. "PERK Is Critical for Alphavirus Nonstructural Protein Translation." Viruses 13, no. 5: 892.
Marine microorganisms have been a resource for novel therapeutic drugs for decades. In addition to anticancer drugs, the drug acyclovir, derived from a marine sponge, is FDA-approved for the treatment of human herpes simplex virus-1 infections. Most alphaviruses that are infectious to terrestrial animals and humans, such as Venezuelan and eastern equine encephalitis viruses (VEEV and EEEV), lack efficient antiviral drugs and it is imperative to develop these remedies. To push the discovery and development of anti-alphavirus compounds forward, this study aimed to isolate and screen for potential antiviral compounds from cultured marine microbes originating from the marine environment. Compounds from marine microbes were of interest as they are prolific producers of bioactive compounds across the spectrum of human diseases and infections. Homoseongomycin, an actinobacteria isolated from a marine sponge displayed impressive activity against VEEV from a total of 76 marine bioactive products. The 50% effective concentration (EC50) for homoseongomycin was 8.6 μM for suppressing VEEV TC-83 luciferase reporter virus replication. Homoseongomycin was non-toxic up to 50 μM and partially rescued cells from VEEV induced cell death. Homoseongomycin exhibited highly efficient antiviral activity with a reduction of VEEV infectious titers by 8 log10 at 50 μM. It also inhibited EEEV replication with an EC50 of 1.2 μM. Mechanism of action studies suggest that homoseongomycin affects both early and late stages of the viral life cycle. Cells treated with 25 μM of homoseongomycin had a ~90% reduction in viral entry. In comparison, later stages showed a more robust reduction in infectious titers (6 log10) and VEEV extracellular viral RNA levels (4 log10), but a lesser impact on intracellular viral RNA levels (1.5 log10). In sum, this work demonstrates that homoseongomycin is a potential anti-VEEV and anti-EEEV compound due to its low cytotoxicity and potent antiviral activity.
Shih-Chao Lin; Caitlin W. Lehman; Allison K. Stewart; Lauren Panny; Nicole Bracci; Jeffrey L.C. Wright; Mikell Paige; Wendy K. Strangman; Kylene Kehn-Hall. Homoseongomycin, a compound isolated from marine actinomycete bacteria K3-1, is a potent inhibitor of encephalitic alphaviruses. Antiviral Research 2021, 191, 105087 .
AMA StyleShih-Chao Lin, Caitlin W. Lehman, Allison K. Stewart, Lauren Panny, Nicole Bracci, Jeffrey L.C. Wright, Mikell Paige, Wendy K. Strangman, Kylene Kehn-Hall. Homoseongomycin, a compound isolated from marine actinomycete bacteria K3-1, is a potent inhibitor of encephalitic alphaviruses. Antiviral Research. 2021; 191 ():105087.
Chicago/Turabian StyleShih-Chao Lin; Caitlin W. Lehman; Allison K. Stewart; Lauren Panny; Nicole Bracci; Jeffrey L.C. Wright; Mikell Paige; Wendy K. Strangman; Kylene Kehn-Hall. 2021. "Homoseongomycin, a compound isolated from marine actinomycete bacteria K3-1, is a potent inhibitor of encephalitic alphaviruses." Antiviral Research 191, no. : 105087.
Alphaviruses are a genus of the Togaviridae family and are widely distributed across the globe. Venezuelan equine encephalitis virus (VEEV) and eastern equine encephalitis virus (EEEV), cause encephalitis and neurological sequelae while chikungunya virus (CHIKV) and Sindbis virus (SINV) cause arthralgia. There are currently no approved therapeutics or vaccines available for alphaviruses. In order to identify novel therapeutics, a V5 epitope tag was inserted into the N-terminus of the VEEV E2 glycoprotein and used to identify host-viral protein interactions. Host proteins involved in protein folding, metabolism/ATP production, translation, cytoskeleton, complement, vesicle transport and ubiquitination were identified as VEEV E2 interactors. Multiple inhibitors targeting these host proteins were tested to determine their effect on VEEV replication. The compound HA15, a GRP78 inhibitor, was found to be an effective inhibitor of VEEV, EEEV, CHIKV, and SINV. VEEV E2 interaction with GRP78 was confirmed through coimmunoprecipitation and colocalization experiments. Mechanism of action studies found that HA15 does not affect viral RNA replication but instead affects late stages of the viral life cycle, which is consistent with GRP78 promoting viral assembly or viral protein trafficking.
Michael Barrera; Victoria Callahan; Ivan Akhrymuk; Nishank Bhalla; Weidong Zhou; Catherine Campbell; Aarthi Narayanan; Kylene Kehn-Hall. Proteomic Discovery of VEEV E2-Host Partner Interactions Identifies GRP78 Inhibitor HA15 as a Potential Therapeutic for Alphavirus Infections. Pathogens 2021, 10, 283 .
AMA StyleMichael Barrera, Victoria Callahan, Ivan Akhrymuk, Nishank Bhalla, Weidong Zhou, Catherine Campbell, Aarthi Narayanan, Kylene Kehn-Hall. Proteomic Discovery of VEEV E2-Host Partner Interactions Identifies GRP78 Inhibitor HA15 as a Potential Therapeutic for Alphavirus Infections. Pathogens. 2021; 10 (3):283.
Chicago/Turabian StyleMichael Barrera; Victoria Callahan; Ivan Akhrymuk; Nishank Bhalla; Weidong Zhou; Catherine Campbell; Aarthi Narayanan; Kylene Kehn-Hall. 2021. "Proteomic Discovery of VEEV E2-Host Partner Interactions Identifies GRP78 Inhibitor HA15 as a Potential Therapeutic for Alphavirus Infections." Pathogens 10, no. 3: 283.
The host proteins Protein Kinase B (AKT) and glycogen synthase kinase-3 (GSK-3) are associated with multiple neurodegenerative disorders. They are also important for the replication of Venezuelan equine encephalitis virus (VEEV), thereby making the AKT/GSK-3 pathway an attractive target for developing anti-VEEV therapeutics. Resveratrol, a natural phytochemical, has been shown to substantially inhibit the AKT pathway. Therefore, we attempted to explore whether it exerts any antiviral activity against VEEV. In this study, we utilized green fluorescent protein (GFP)- and luciferase-encoding recombinant VEEV to determine the cytotoxicity and antiviral efficacy via luciferase reporter assays, flow cytometry, and immunofluorescent assays. Our results indicate that resveratrol treatment is capable of inhibiting VEEV replication, resulting in increased viability of Vero and U87MG cells as well as reduced virion production and viral RNA contents within host cells for at least 48 h with a single treatment. Furthermore, the suppression of apoptotic signaling adaptors, caspase-3, caspase-7, and annexin V may also be implicated in resveratrol-mediated antiviral activity. We found that decreased phosphorylation of the AKT/GSK-3 pathway, mediated by resveratrol, can be triggered during the early stages of VEEV infection, suggesting that resveratrol disrupts the viral replication cycle and consequently promotes cell survival. Finally, molecular docking and dynamics simulation studies revealed that resveratrol can directly bind to VEEV glycoproteins, which may interfere with virus attachment and entry. In conclusion, our results suggest that resveratrol exerts inhibitory activity against VEEV infection and upon further modification could be a useful compound to study in neuroprotective research and veterinary sciences.
Caitlin Lehman; Kylene Kehn-Hall; Megha Aggarwal; Nicole Bracci; Han-Chi Pan; Lauren Panny; Robert Lamb; Shih-Chao Lin. Resveratrol Inhibits Venezuelan Equine Encephalitis Virus Infection by Interfering with the AKT/GSK Pathway. Plants 2021, 10, 346 .
AMA StyleCaitlin Lehman, Kylene Kehn-Hall, Megha Aggarwal, Nicole Bracci, Han-Chi Pan, Lauren Panny, Robert Lamb, Shih-Chao Lin. Resveratrol Inhibits Venezuelan Equine Encephalitis Virus Infection by Interfering with the AKT/GSK Pathway. Plants. 2021; 10 (2):346.
Chicago/Turabian StyleCaitlin Lehman; Kylene Kehn-Hall; Megha Aggarwal; Nicole Bracci; Han-Chi Pan; Lauren Panny; Robert Lamb; Shih-Chao Lin. 2021. "Resveratrol Inhibits Venezuelan Equine Encephalitis Virus Infection by Interfering with the AKT/GSK Pathway." Plants 10, no. 2: 346.
P-selectin glycoprotein ligand-1 (PSGL-1) is a cell surface glycoprotein that binds to P-, E-, and L-selectins to mediate the tethering and rolling of immune cells on the surface of the endothelium for cell migration into inflamed tissues. PSGL-1 has been identified as an interferon-γ (INF-γ)-regulated factor that restricts HIV-1 infectivity, and has recently been found to possess broad-spectrum antiviral activities. Here we report that the expression of PSGL-1 in virus-producing cells impairs the incorporation of SARS-CoV and SARS-CoV-2 spike (S) glycoproteins into pseudovirions and blocks pseudovirus attachment and infection of target cells. These findings suggest that PSGL-1 may potentially inhibit coronavirus replication in PSGL-1+ cells
Sijia He; Abdul A. Waheed; Brian Hetrick; Deemah Dabbagh; Ivan V. Akhrymuk; Kylene Kehn-Hall; Eric O. Freed; Yuntao Wu. PSGL-1 Inhibits the Incorporation of SARS-CoV and SARS-CoV-2 Spike Glycoproteins into Pseudovirions and Impairs Pseudovirus Attachment and Infectivity. Viruses 2020, 13, 46 .
AMA StyleSijia He, Abdul A. Waheed, Brian Hetrick, Deemah Dabbagh, Ivan V. Akhrymuk, Kylene Kehn-Hall, Eric O. Freed, Yuntao Wu. PSGL-1 Inhibits the Incorporation of SARS-CoV and SARS-CoV-2 Spike Glycoproteins into Pseudovirions and Impairs Pseudovirus Attachment and Infectivity. Viruses. 2020; 13 (1):46.
Chicago/Turabian StyleSijia He; Abdul A. Waheed; Brian Hetrick; Deemah Dabbagh; Ivan V. Akhrymuk; Kylene Kehn-Hall; Eric O. Freed; Yuntao Wu. 2020. "PSGL-1 Inhibits the Incorporation of SARS-CoV and SARS-CoV-2 Spike Glycoproteins into Pseudovirions and Impairs Pseudovirus Attachment and Infectivity." Viruses 13, no. 1: 46.
In light of the COVID-19 pandemic, studies that work to understand SARS-CoV-2 are urgently needed. In turn, the less severe human coronaviruses such as HCoV-229E and OC43 are drawing newfound attention. These less severe coronaviruses can be used as a model to facilitate our understanding of the host immune response to coronavirus infection. SARS-CoV-2 must be handled under biosafety level 3 (BSL-3) conditions. Therefore, HCoV-229E and OC43, which can be handled at BSL-2 provide an alternative to SARS-CoV-2 for preclinical screening and designing of antivirals. However, to date, there is no published effective and efficient method to titrate HCoVs other than expensive indirect immunostaining. Here we present an improved approach using an agarose-based conventional plaque assay to titrate HCoV 229E and OC43 with mink lung epithelial cells, Mv1Lu. Our results indicate that titration of HCoV 229E and OC43 with Mv1Lu is consistent and reproducible. The titers produced are also comparable to those produced using human rhabdomyosarcoma (RD) cells. More importantly, Mv1Lu cells display a higher tolerance for cell-cell contact stress, decreased temperature sensitivity, and a faster growth rate. We believe that our improved low-cost plaque assay can serve as an easy tool for researchers conducting HCoV research.
Nicole Bracci; Han-Chi Pan; Caitlin Lehman; Kylene Kehn-Hall; Shih-Chao Lin. Improved plaque assay for human coronaviruses 229E and OC43. PeerJ 2020, 8, e10639 .
AMA StyleNicole Bracci, Han-Chi Pan, Caitlin Lehman, Kylene Kehn-Hall, Shih-Chao Lin. Improved plaque assay for human coronaviruses 229E and OC43. PeerJ. 2020; 8 ():e10639.
Chicago/Turabian StyleNicole Bracci; Han-Chi Pan; Caitlin Lehman; Kylene Kehn-Hall; Shih-Chao Lin. 2020. "Improved plaque assay for human coronaviruses 229E and OC43." PeerJ 8, no. : e10639.
The coronavirus disease 2019 (COVID-19) has created an acute worldwide demand for sustained broadband pathogen suppression in households, hospitals, and public spaces. The US recently passed a new sad milestone of 500,000 deaths due to COVID-19, the highest rate anywhere in the world. In response, we have created a rapid-acting, self-sterilizing PPE configurations capable of killing SARS-CoV-2 and other microbes in seconds. The highly active material destroys pathogens faster than any conventional copper configuration. The material maintains its antimicrobial efficacy over sustained use and is shelf stable. We have performed rigorous testing in accordance with guidelines from U.S. governing authorities and believe that the material could offer broad spectrum, non-selective defense against most microbes via integration into masks and other protective equipment.
Alfred A. Zinn; Mina Izadjoo; Hosan Kim; Kylene Kehn-Hall; Caitlin Lehman; Rachel L. Brody; Robert R. Roth; Augustin Vega; Khanh K. Nguyen; Nhi T. Ngo; Hannah T. Zinn; Lauren Panny; Rafaela Flor; Nicholas Antonopoulos; Randall M. Stoltenberg. Rapidly self-sterilizing PPE capable of 99.9% SARS-CoV-2 deactivation in 30 seconds. 2020, 1 .
AMA StyleAlfred A. Zinn, Mina Izadjoo, Hosan Kim, Kylene Kehn-Hall, Caitlin Lehman, Rachel L. Brody, Robert R. Roth, Augustin Vega, Khanh K. Nguyen, Nhi T. Ngo, Hannah T. Zinn, Lauren Panny, Rafaela Flor, Nicholas Antonopoulos, Randall M. Stoltenberg. Rapidly self-sterilizing PPE capable of 99.9% SARS-CoV-2 deactivation in 30 seconds. . 2020; ():1.
Chicago/Turabian StyleAlfred A. Zinn; Mina Izadjoo; Hosan Kim; Kylene Kehn-Hall; Caitlin Lehman; Rachel L. Brody; Robert R. Roth; Augustin Vega; Khanh K. Nguyen; Nhi T. Ngo; Hannah T. Zinn; Lauren Panny; Rafaela Flor; Nicholas Antonopoulos; Randall M. Stoltenberg. 2020. "Rapidly self-sterilizing PPE capable of 99.9% SARS-CoV-2 deactivation in 30 seconds." , no. : 1.
The emergence of SARS-CoV-2 has created an international health crisis. Small animal models mirroring SARS-CoV-2 human disease are essential for medical countermeasure (MCM) development. Mice are refractory to SARS-CoV-2 infection due to low affinity binding to the murine angiotensin-converting enzyme 2 (ACE2) protein. Here we evaluated the pathogenesis of SARS-CoV-2 in male and female mice expressing the human ACE2 gene under the control of the keratin 18 promotor. In contrast to non-transgenic mice, intranasal exposure of K18-hACE2 animals to two different doses of SARS-CoV-2 resulted in acute disease including weight loss, lung injury, brain infection and lethality. Vasculitis was the most prominent finding in the lungs of infected mice. Transcriptomic analysis from lungs of infected animals revealed increases in transcripts involved in lung injury and inflammatory cytokines. In the lower dose challenge groups, there was a survival advantage in the female mice with 60% surviving infection whereas all male mice succumbed to disease. Male mice that succumbed to disease had higher levels of inflammatory transcripts compared to female mice. This is the first highly lethal murine infection model for SARS-CoV-2. The K18-hACE2 murine model will be valuable for the study of SARS-CoV-2 pathogenesis and the assessment of MCMs.
Joseph W. Golden; Curtis R. Cline; Xiankun Zeng; Aura R. Garrison; Brian D. Carey; Eric M. Mucker; Lauren E. White; Joshua D. Shamblin; Rebecca L. Brocato; Jun Liu; April M. Babka; Hypaitia B. Rauch; Jeffrey M. Smith; Bradley S. Hollidge; Collin Fitzpatrick; Catherine V. Badger; Jay W. Hooper. Human angiotensin-converting enzyme 2 transgenic mice infected with SARS-CoV-2 develop severe and fatal respiratory disease. JCI Insight 2020, 5, 1 .
AMA StyleJoseph W. Golden, Curtis R. Cline, Xiankun Zeng, Aura R. Garrison, Brian D. Carey, Eric M. Mucker, Lauren E. White, Joshua D. Shamblin, Rebecca L. Brocato, Jun Liu, April M. Babka, Hypaitia B. Rauch, Jeffrey M. Smith, Bradley S. Hollidge, Collin Fitzpatrick, Catherine V. Badger, Jay W. Hooper. Human angiotensin-converting enzyme 2 transgenic mice infected with SARS-CoV-2 develop severe and fatal respiratory disease. JCI Insight. 2020; 5 (19):1.
Chicago/Turabian StyleJoseph W. Golden; Curtis R. Cline; Xiankun Zeng; Aura R. Garrison; Brian D. Carey; Eric M. Mucker; Lauren E. White; Joshua D. Shamblin; Rebecca L. Brocato; Jun Liu; April M. Babka; Hypaitia B. Rauch; Jeffrey M. Smith; Bradley S. Hollidge; Collin Fitzpatrick; Catherine V. Badger; Jay W. Hooper. 2020. "Human angiotensin-converting enzyme 2 transgenic mice infected with SARS-CoV-2 develop severe and fatal respiratory disease." JCI Insight 5, no. 19: 1.
Introduction Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which is responsible for the COVID-19 pandemic, remains viable and therefore potentially infectious on several materials. One strategy to discourage the fomite-mediated spread of COVID-19 is the development of materials whose surface chemistry can spontaneously inactivate SARS-CoV-2. Silicon nitride (Si3N4), a material used in spine fusion surgery, is one such candidate because it has been shown to inactivate several bacterial species and viral strains. This study hypothesized that contact with Si3N4would inactivate SARS-CoV-2, while mammalian cells would remain unaffected. Materials SARS-CoV-2 virions (2×104PFU/mL diluted in growth media) were exposed to 5, 10, 15, and 20% (w/v) of an aqueous suspension of sintered Si3N4particles for durations of 1, 5, and 10 minutes, respectively. Before exposure to the virus, cytotoxicity testing of Si3N4alone was assessed in Vero cells at 24 and 48 hour post-exposure times. Following each exposure to Si3N4, the remaining infectious virus was quantitated by plaque assay. Results Vero cell viability increased at 5% and 10% (w/v) concentrations of Si3N4at exposure times up to 10 minutes, and there was only minimal impact on cell health and viability up to 20% (w/v). However, the SARS-CoV-2 titers were markedly reduced when exposed to all concentrations of Si3N4; the reduction in viral titers was between 85% - 99.6%, depending on the dose and duration of exposure. Conclusions Si3N4was non-toxic to the Vero cells while showing strong antiviral activity against SARS-CoV-2. The viricidal effect increased with increasing concentrations of Si3N4and longer duration of exposure. Surface treatment strategies based on Si3N4may offer novel methods to discourage SARS-CoV-2 persistence and infectivity on surfaces and discourage the spread of COVID-19.
Caitlin W. Lehman; Rafaela Flur; Kylene Kehn-Hall; Bryan J. McEntire; B. Sonny Bal; Ryan M. Bock. Silicon Nitride Inactivates SARS-CoV-2in vitro. 2020, 1 .
AMA StyleCaitlin W. Lehman, Rafaela Flur, Kylene Kehn-Hall, Bryan J. McEntire, B. Sonny Bal, Ryan M. Bock. Silicon Nitride Inactivates SARS-CoV-2in vitro. . 2020; ():1.
Chicago/Turabian StyleCaitlin W. Lehman; Rafaela Flur; Kylene Kehn-Hall; Bryan J. McEntire; B. Sonny Bal; Ryan M. Bock. 2020. "Silicon Nitride Inactivates SARS-CoV-2in vitro." , no. : 1.
Coronavirus disease 2019 (COVID-19), caused by the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), became a pandemic in early 2020. Lateral flow immunoassays for antibody testing have been viewed as a cheap and rapidly deployable method for determining previous infection with SARS-CoV-2; however, these assays have shown unacceptably low sensitivity. We report on nine lateral flow immunoassays currently available and compare their titer sensitivity in serum to a best-practice enzyme-linked immunosorbent assay (ELISA) and viral neutralization assay. For a small group of PCR-positive, we found two lateral flow immunoassay devices with titer sensitivity roughly equal to the ELISA; these devices were positive for all PCR-positive patients harboring SARS-CoV-2 neutralizing antibodies. One of these devices was deployed in Northern Italy to test its sensitivity and specificity in a real-world clinical setting. Using the device with fingerstick blood on a cohort of 27 hospitalized PCR-positive patients and seven hospitalized controls, ROC curve analysis gave AUC values of 0.7646 for IgG. For comparison, this assay was also tested with saliva from the same patient population and showed reduced discrimination between cases and controls with AUC values of 0.6841 for IgG. Furthermore, during viral neutralization testing, one patient was discovered to harbor autoantibodies to ACE2, with implications for how immune responses are profiled. We show here through a proof-of-concept study that these lateral flow devices can be as analytically sensitive as ELISAs and adopted into hospital protocols; however, additional improvements to these devices remain necessary before their clinical deployment.
Amanda Haymond; Claudius Mueller; Hannah Steinberg; K. Alex Hodge; Caitlin W Lehman; Shih-Chao Lin; Lucia Collini; Heather Branscome; Tuong Vi Nguyen; Sally Rucker; Lauren Panny; Rafaela Flor; Raouf Guirguis; Richard Hoefer; Giovanni Lorenzin; Emanuel Petricoin; Fatah Kashanchi; Kylene Kehn-Hall; Paolo Lanzafame; Lance Liotta; Alessandra Luchini. Clinical Utility of a Highly Sensitive Lateral Flow Immunoassay as determined by Titer Analysis for the Detection of anti-SARS-CoV-2 Antibodies at the Point-of-Care. 2020, 1 .
AMA StyleAmanda Haymond, Claudius Mueller, Hannah Steinberg, K. Alex Hodge, Caitlin W Lehman, Shih-Chao Lin, Lucia Collini, Heather Branscome, Tuong Vi Nguyen, Sally Rucker, Lauren Panny, Rafaela Flor, Raouf Guirguis, Richard Hoefer, Giovanni Lorenzin, Emanuel Petricoin, Fatah Kashanchi, Kylene Kehn-Hall, Paolo Lanzafame, Lance Liotta, Alessandra Luchini. Clinical Utility of a Highly Sensitive Lateral Flow Immunoassay as determined by Titer Analysis for the Detection of anti-SARS-CoV-2 Antibodies at the Point-of-Care. . 2020; ():1.
Chicago/Turabian StyleAmanda Haymond; Claudius Mueller; Hannah Steinberg; K. Alex Hodge; Caitlin W Lehman; Shih-Chao Lin; Lucia Collini; Heather Branscome; Tuong Vi Nguyen; Sally Rucker; Lauren Panny; Rafaela Flor; Raouf Guirguis; Richard Hoefer; Giovanni Lorenzin; Emanuel Petricoin; Fatah Kashanchi; Kylene Kehn-Hall; Paolo Lanzafame; Lance Liotta; Alessandra Luchini. 2020. "Clinical Utility of a Highly Sensitive Lateral Flow Immunoassay as determined by Titer Analysis for the Detection of anti-SARS-CoV-2 Antibodies at the Point-of-Care." , no. : 1.
Protein phosphorylation plays an important role during the life cycle of many viruses. Venezuelan equine encephalitis virus (VEEV) capsid protein has recently been shown to be phosphorylated at four residues. Here those studies are extended to determine the kinase responsible for phosphorylation and the importance of capsid phosphorylation during the viral life cycle. Phosphorylation site prediction software suggests that Protein Kinase C (PKC) is responsible for phosphorylation of VEEV capsid. VEEV capsid co-immunoprecipitated with PKCδ, but not other PKC isoforms and siRNA knockdown of PKCδ caused a decrease in viral replication. Furthermore, knockdown of PKCδ by siRNA decreased capsid phosphorylation. A virus with capsid phosphorylation sites mutated to alanine (VEEV CPD) displayed a lower genomic copy to pfu ratio than the parental virus; suggesting more efficient viral assembly and more infectious particles being released. RNA:capsid binding was significantly increased in the mutant virus, confirming these results. Finally, VEEV CPD is attenuated in a mouse model of infection, with mice showing increased survival and decreased clinical signs as compared to mice infected with the parental virus. Collectively our data support a model in which PKCδ mediated capsid phosphorylation regulates viral RNA binding and assembly, significantly impacting viral pathogenesis.
Brian D. Carey; Ivan Akhrymuk; Bibha Dahal; Chelsea L. Pinkham; Nicole Bracci; Sarah Finstuen-Magro; Shih-Chao Lin; Caitlin W. Lehman; Kevin J. Sokoloski; Kylene Kehn-Hall. Protein Kinase C subtype δ interacts with Venezuelan equine encephalitis virus capsid protein and regulates viral RNA binding through modulation of capsid phosphorylation. PLOS Pathogens 2020, 16, e1008282 .
AMA StyleBrian D. Carey, Ivan Akhrymuk, Bibha Dahal, Chelsea L. Pinkham, Nicole Bracci, Sarah Finstuen-Magro, Shih-Chao Lin, Caitlin W. Lehman, Kevin J. Sokoloski, Kylene Kehn-Hall. Protein Kinase C subtype δ interacts with Venezuelan equine encephalitis virus capsid protein and regulates viral RNA binding through modulation of capsid phosphorylation. PLOS Pathogens. 2020; 16 (3):e1008282.
Chicago/Turabian StyleBrian D. Carey; Ivan Akhrymuk; Bibha Dahal; Chelsea L. Pinkham; Nicole Bracci; Sarah Finstuen-Magro; Shih-Chao Lin; Caitlin W. Lehman; Kevin J. Sokoloski; Kylene Kehn-Hall. 2020. "Protein Kinase C subtype δ interacts with Venezuelan equine encephalitis virus capsid protein and regulates viral RNA binding through modulation of capsid phosphorylation." PLOS Pathogens 16, no. 3: e1008282.
Nanotrap® (NT) particles are hydrogel microspheres developed for target analyte separation and discovery applications. NT particles consist of cross-linked N-isopropylacrylamide (NIPAm) copolymers that are functionalized with a variety of chemical affinity baits to enable broad-spectrum collection and retention of target proteins, nucleic acids, and pathogens. NT particles have been previously shown to capture and enrich arboviruses including Rift Valley fever and Venezuelan equine encephalitis viruses. Yet, there is still a need to enhance the detection ability for other re-emerging viruses such as Zika (ZIKV), chikungunya (CHIKV), and dengue (DENV) viruses. In this study, we exploited NT particles with different affinity baits, including cibacron blue, acrylic acid, and reactive red 120, to evaluate their capturing and enrichment capability for ZIKV, DENV and CHIKV in human fluids. Our results demonstrate that CN1030, a NT particle conjugated with reactive red 120, can recover between 8-16-fold greater genomic copies of ZIKV, CHIKV and DENV in virus spiked urine samples via RT-qPCR, superior to the other chemical baits. Also, we observed that CN1030 simultaneously enriched ZIKV, CHIKV and DENV in co-infection-based settings and could stabilize ZIKV, but not CHIKV infectivity in saliva spiked samples. CN1030 enriched viral detection at various viral concentrations, with significant enhancement observed at viral titers as low as 100 PFU/mL for ZIKV and 10 PFU/mL for CHIKV. The detection of ZIKV was further enhanced with NT particles by processing of larger volume urine samples. Furthermore, we developed a magnetic NT particle, CN3080, based on the same backbone of CN1030, and demonstrated that CN3080 could also capture and enrich ZIKV and CHIKV in a dose-dependent manner. Finally, in silico docking predictions support that the affinity between reactive red 120 and ZIKV or CHIKV envelope proteins appeared to be greater than acrylic acid. Overall, our data show that NT particles along with reactive red 120 can be utilized as a pre-processing technology for enhancement of detecting febrile-illness causing viruses.
Shih-Chao Lin; Brian D. Carey; Victoria Callahan; Ji-Hyun Lee; Nicole Bracci; Anurag Patnaik; Amy K. Smith; Aarthi Narayanan; Benjamin Lepene; Kylene Kehn-Hall. Use of Nanotrap particles for the capture and enrichment of Zika, chikungunya and dengue viruses in urine. PLOS ONE 2020, 15, e0227058 .
AMA StyleShih-Chao Lin, Brian D. Carey, Victoria Callahan, Ji-Hyun Lee, Nicole Bracci, Anurag Patnaik, Amy K. Smith, Aarthi Narayanan, Benjamin Lepene, Kylene Kehn-Hall. Use of Nanotrap particles for the capture and enrichment of Zika, chikungunya and dengue viruses in urine. PLOS ONE. 2020; 15 (1):e0227058.
Chicago/Turabian StyleShih-Chao Lin; Brian D. Carey; Victoria Callahan; Ji-Hyun Lee; Nicole Bracci; Anurag Patnaik; Amy K. Smith; Aarthi Narayanan; Benjamin Lepene; Kylene Kehn-Hall. 2020. "Use of Nanotrap particles for the capture and enrichment of Zika, chikungunya and dengue viruses in urine." PLOS ONE 15, no. 1: e0227058.
Venezuelan equine encephalitis virus (VEEV) is a neurotropic virus that causes significant disease in both humans and equines. Here we characterized the impact of VEEV on signaling pathways regulating cell death in human primary astrocytes. VEEV productively infected primary astrocytes and caused an upregulation of early growth response 1 (EGR1) gene expression at 9 and 18 h post infection. EGR1 induction was dependent on extracellular signal-regulated kinase1/2 (ERK1/2) and protein kinase R (PKR)-like endoplasmic reticulum kinase (PERK), but not on p38 mitogen activated protein kinase (MAPK) or phosphoinositide 3-kinase (PI3K) signaling. Knockdown of EGR1 significantly reduced VEEV-induced apoptosis and impacted viral replication. Knockdown of ERK1/2 or PERK significantly reduced EGR1 gene expression, dramatically reduced viral replication, and increased cell survival as well as rescued cells from VEEV-induced apoptosis. These data indicate that EGR1 activation and subsequent cell death are regulated through ERK and PERK pathways in VEEV infected primary astrocytes.
Bibha Dahal; Shih-Chao Lin; Brian D. Carey; Jonathan L. Jacobs; Jonathan Dinman; Monique van Hoek; Andre A. Adams; Kylene Kehn-Hall. EGR1 upregulation following Venezuelan equine encephalitis virus infection is regulated by ERK and PERK pathways contributing to cell death. Virology 2019, 539, 121 -128.
AMA StyleBibha Dahal, Shih-Chao Lin, Brian D. Carey, Jonathan L. Jacobs, Jonathan Dinman, Monique van Hoek, Andre A. Adams, Kylene Kehn-Hall. EGR1 upregulation following Venezuelan equine encephalitis virus infection is regulated by ERK and PERK pathways contributing to cell death. Virology. 2019; 539 ():121-128.
Chicago/Turabian StyleBibha Dahal; Shih-Chao Lin; Brian D. Carey; Jonathan L. Jacobs; Jonathan Dinman; Monique van Hoek; Andre A. Adams; Kylene Kehn-Hall. 2019. "EGR1 upregulation following Venezuelan equine encephalitis virus infection is regulated by ERK and PERK pathways contributing to cell death." Virology 539, no. : 121-128.
A tertiary structure governs, to a great extent, the biological activity of a protein in the living cell and is consequently a central focus of numerous studies aiming to shed light on cellular processes central to human health. Here, we aim to elucidate the structure of the Rift Valley fever virus (RVFV) L protein using a combination of in silico techniques. Due to its large size and multiple domains, elucidation of the tertiary structure of the L protein has so far challenged both dry and wet laboratories. In this work, we leverage complementary perspectives and tools from the computational-molecular-biology and bioinformatics domains for constructing, refining, and evaluating several atomistic structural models of the L protein that are physically realistic. All computed models have very flexible termini of about 200 amino acids each, and a high proportion of helical regions. Properties such as potential energy, radius of gyration, hydrodynamics radius, flexibility coefficient, and solvent-accessible surface are reported. Structural characterization of the L protein enables our laboratories to better understand viral replication and transcription via further studies of L protein-mediated protein–protein interactions. While results presented a focus on the RVFV L protein, the following workflow is a more general modeling protocol for discovering the tertiary structure of multidomain proteins consisting of thousands of amino acids.
Gideon K. Gogovi; Fahad Almsned; Nicole Bracci; Kylene Kehn-Hall; Amarda Shehu; Estela Blaisten-Barojas. Modeling the Tertiary Structure of the Rift Valley Fever Virus L Protein. Molecules 2019, 24, 1768 .
AMA StyleGideon K. Gogovi, Fahad Almsned, Nicole Bracci, Kylene Kehn-Hall, Amarda Shehu, Estela Blaisten-Barojas. Modeling the Tertiary Structure of the Rift Valley Fever Virus L Protein. Molecules. 2019; 24 (9):1768.
Chicago/Turabian StyleGideon K. Gogovi; Fahad Almsned; Nicole Bracci; Kylene Kehn-Hall; Amarda Shehu; Estela Blaisten-Barojas. 2019. "Modeling the Tertiary Structure of the Rift Valley Fever Virus L Protein." Molecules 24, no. 9: 1768.
Zika virus (ZIKV) is a re-emerging Flavivirus that has been linked to microcephaly and other neurological pathologies. In this study, phloretin, a glucose transporter inhibitor naturally derived from plants, was used to investigate the glucose dependence of ZIKV replication in host cells. The results showed that phloretin significantly decreased infectious titres of two ZIKV strains, namely MR766 (African genotype) and PRVABC59 (Puerto Rico genotype). The 50% effective concentration (EC50) of phloretin against MR766 and PRVABC59 was 22.85 µM and 9.31 µM, respectively. Further analyses demonstrated that decreased viral production was due to host-targeted inhibition, including decreased apoptotic caspase-3 and -7 activities and reduced phosphorylation of Akt/mTOR pathways. In addition, upon disruption of cellular glucose availability within host cells using 2-deoxy-d-glucose, ZIKV propagation was inhibited. Collectively, we demonstrate phloretin inhibition of ZIKV propagation and provide evidence of glucose utilization pathways as being important for ZIKV propagation. The activity of phloretin and its role in inhibiting glucose uptake could provide a useful foundation for the development of ZIKV antivirals.
Shih-Chao Lin; Mei-Chun Chen; Shufeng Liu; Victoria M. Callahan; Nicole R. Bracci; Caitlin W. Lehman; Bibha Dahal; Cynthia L. de la Fuente; Chi-Chen Lin; Tony T. Wang; Kylene Kehn-Hall. Phloretin inhibits Zika virus infection by interfering with cellular glucose utilisation. International Journal of Antimicrobial Agents 2019, 54, 80 -84.
AMA StyleShih-Chao Lin, Mei-Chun Chen, Shufeng Liu, Victoria M. Callahan, Nicole R. Bracci, Caitlin W. Lehman, Bibha Dahal, Cynthia L. de la Fuente, Chi-Chen Lin, Tony T. Wang, Kylene Kehn-Hall. Phloretin inhibits Zika virus infection by interfering with cellular glucose utilisation. International Journal of Antimicrobial Agents. 2019; 54 (1):80-84.
Chicago/Turabian StyleShih-Chao Lin; Mei-Chun Chen; Shufeng Liu; Victoria M. Callahan; Nicole R. Bracci; Caitlin W. Lehman; Bibha Dahal; Cynthia L. de la Fuente; Chi-Chen Lin; Tony T. Wang; Kylene Kehn-Hall. 2019. "Phloretin inhibits Zika virus infection by interfering with cellular glucose utilisation." International Journal of Antimicrobial Agents 54, no. 1: 80-84.
The New World alphaviruses, Venezuelan, eastern and western equine encephalitis viruses (VEEV, EEEV, and WEEV), are important human pathogens due to their ability to cause varying levels of morbidity and mortality in humans. There is also concern about VEEV and EEEV being used as bioweapons. Currently, a FDA-approved antiviral is lacking for New World alphaviruses. In this review, the function of each viral protein is discussed with an emphasis on how these functions can be targeted by therapeutics. Both direct acting antivirals as well as inhibitors that impact host protein interactions with viral proteins are described. Non-structural protein 3 (nsP3), capsid, and E2 proteins have garnered attention in recent years, whereas little is known regarding host protein interactions of the other viral proteins and is an important avenue for future study.
Brian D Carey; Allison Bakovic; Victoria Callahan; Aarthi Narayanan; Kylene Kehn-Hall. New World alphavirus protein interactomes from a therapeutic perspective. Antiviral Research 2019, 163, 125 -139.
AMA StyleBrian D Carey, Allison Bakovic, Victoria Callahan, Aarthi Narayanan, Kylene Kehn-Hall. New World alphavirus protein interactomes from a therapeutic perspective. Antiviral Research. 2019; 163 ():125-139.
Chicago/Turabian StyleBrian D Carey; Allison Bakovic; Victoria Callahan; Aarthi Narayanan; Kylene Kehn-Hall. 2019. "New World alphavirus protein interactomes from a therapeutic perspective." Antiviral Research 163, no. : 125-139.
Venezuelan equine encephalitis virus (VEEV) is a positive sense, single-stranded RNA virus and member of the New World alphaviruses. It causes a biphasic febrile illness that can be accompanied by central nervous system involvement and moderate morbidity in humans and severe mortality in equines. The virus has a history of weaponization, lacks FDA-approved therapeutics and vaccines in humans, and is considered a select agent. Like other RNA viruses, VEEV replicates in the cytoplasm of infected cells and eventually induces apoptosis. The capsid protein, which contains a nuclear localization and a nuclear export sequence, induces a shutdown of host transcription and nucleocytoplasmic trafficking. Here we show that infection with VEEV causes a dysregulation of cell cycling and a delay in the G0/G1 phase in Vero cells and U87MG astrocytes. Cells infected with VEEV encoding a capsid NLS mutant or treated with the capsid-importin α interaction inhibitor G281-1485 were partially rescued from this cell cycle dysregulation. Pathway analysis of previously published RNA-sequencing data from VEEV infected U87MG astrocytes identified alterations of canonical pathways involving cell cycle, checkpoint regulation, and proliferation. Multiple cyclins including cyclin D1, cyclin A2 and cyclin E2 and other regulators of the cell cycle were downregulated in infected cells in a capsid NLS dependent manner. Loss of Rb phosphorylation, which is a substrate for cyclin/cdk complexes was also observed. These data demonstrate the importance of capsid nuclear localization and/or importin α binding for inducing cell cycle arrest and transcriptional downregulation of key cell cycle regulators.
Lindsay Lundberg; Jacque Fontenot; Shih-Chao Lin; Chelsea Pinkham; Brian D Carey; Catherine E. Campbell; Kylene Kehn-Hall. Venezuelan Equine Encephalitis Virus Capsid Implicated in Infection-Induced Cell Cycle Delay in vitro. Frontiers in Microbiology 2018, 9, 3126 .
AMA StyleLindsay Lundberg, Jacque Fontenot, Shih-Chao Lin, Chelsea Pinkham, Brian D Carey, Catherine E. Campbell, Kylene Kehn-Hall. Venezuelan Equine Encephalitis Virus Capsid Implicated in Infection-Induced Cell Cycle Delay in vitro. Frontiers in Microbiology. 2018; 9 ():3126.
Chicago/Turabian StyleLindsay Lundberg; Jacque Fontenot; Shih-Chao Lin; Chelsea Pinkham; Brian D Carey; Catherine E. Campbell; Kylene Kehn-Hall. 2018. "Venezuelan Equine Encephalitis Virus Capsid Implicated in Infection-Induced Cell Cycle Delay in vitro." Frontiers in Microbiology 9, no. : 3126.
Rift Valley fever virus (RVFV) is an enveloped, segmented, negative sense RNA virus that replicates within the host's cytoplasm. To facilitate its replication, RVFV must utilize host cell processes and as such, these processes may serve as potential therapeutic targets. This review summarizes key host cell processes impacted by RVFV infection. Specifically the influence of RVFV on host transcriptional regulation, post-transcriptional regulation, protein half-life and availability, host signal transduction, trafficking and secretory pathways, cytoskeletal modulation, and mitochondrial processes and oxidative stress are discussed. Therapeutics targeted towards host processes that are essential for RVFV to thrive as well as their efficacy and importance to viral pathogenesis is highlighted.
Chelsea Pinkham; Aslaa Ahmed; Nicole Bracci; Aarthi Narayanan; Kylene Kehn-Hall. Host-based processes as therapeutic targets for Rift Valley fever virus. Antiviral Research 2018, 160, 64 -78.
AMA StyleChelsea Pinkham, Aslaa Ahmed, Nicole Bracci, Aarthi Narayanan, Kylene Kehn-Hall. Host-based processes as therapeutic targets for Rift Valley fever virus. Antiviral Research. 2018; 160 ():64-78.
Chicago/Turabian StyleChelsea Pinkham; Aslaa Ahmed; Nicole Bracci; Aarthi Narayanan; Kylene Kehn-Hall. 2018. "Host-based processes as therapeutic targets for Rift Valley fever virus." Antiviral Research 160, no. : 64-78.