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Red cells can be labelled with peptides from the SARS-CoV-2 spike protein and used for serologic screening of SARS-CoV-2 antibodies. We evaluated 140 convalescent COVID-19 patients and 275 healthy controls using this C19-kodecyte assay. The analytical performance of the new assay was compared with a virus neutralizing assay and 2 commercial chemiluminescent antibody tests (Total assay and IgG assay, Ortho). The C19-kodecyte assay detected SARS-CoV-2 antibodies with a sensitivity of 92.8% and specificity of 96.3%, well within the minimum performance range required by FDA for EUA authorization of serologic tests. The Cohen's kappa coefficient was 0.90 indicating an almost perfect agreement with the Total assay. The Pearson correlation coefficient was 0.20 with the neutralizing assay (0.49 with IgG, and 0.41 with Total assays). The limited correlation in assay reaction strengths suggested that the assays may detect different antibody specificities. Our easily scalable C19-kodecyte assay may vastly improve test capacity in blood typing laboratories using their routine setups for column agglutination technique.
Kshitij Srivastava; Kamille A West; Valeria De Giorgi; Michael R Holbrook; Nicolai V. Bovin; Stephen M Henry; Willy A Flegel. COVID-19 antibody detection and assay performance using red cell agglutination. 2021, 1 .
AMA StyleKshitij Srivastava, Kamille A West, Valeria De Giorgi, Michael R Holbrook, Nicolai V. Bovin, Stephen M Henry, Willy A Flegel. COVID-19 antibody detection and assay performance using red cell agglutination. . 2021; ():1.
Chicago/Turabian StyleKshitij Srivastava; Kamille A West; Valeria De Giorgi; Michael R Holbrook; Nicolai V. Bovin; Stephen M Henry; Willy A Flegel. 2021. "COVID-19 antibody detection and assay performance using red cell agglutination." , no. : 1.
As the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic expanded, it was clear that effective testing for the presence of neutralizing antibodies in the blood of convalescent patients would be critical for development of plasma-based therapeutic approaches. To address the need for a high-quality neutralization assay against SARS-CoV-2, a previously established fluorescence reduction neutralization assay (FRNA) against Middle East respiratory syndrome coronavirus (MERS-CoV) was modified and optimized. The SARS-CoV-2 FRNA provides a quantitative assessment of a large number of infected cells through use of a high-content imaging system. Because of this approach, and the fact that it does not involve subjective interpretation, this assay is more efficient and more accurate than other neutralization assays. In addition, the ability to set robust acceptance criteria for individual plates and specific test wells provided further rigor to this assay. Such agile adaptability avails use with multiple virus variants. By February 2021, the SARS-CoV-2 FRNA had been used to screen over 5000 samples, including acute and convalescent plasma or serum samples and therapeutic antibody treatments, for SARS-CoV-2 neutralizing titers.
Richard Bennett; Elena Postnikova; Janie Liang; Robin Gross; Steven Mazur; Saurabh Dixit; Gregory Kocher; Shuiqing Yu; Shalamar Georgia-Clark; Dawn Gerhardt; Yingyun Cai; Lindsay Marron; Vladimir Lukin; Michael Holbrook. Scalable, Micro-Neutralization Assay for Assessment of SARS-CoV-2 (COVID-19) Virus-Neutralizing Antibodies in Human Clinical Samples. Viruses 2021, 13, 893 .
AMA StyleRichard Bennett, Elena Postnikova, Janie Liang, Robin Gross, Steven Mazur, Saurabh Dixit, Gregory Kocher, Shuiqing Yu, Shalamar Georgia-Clark, Dawn Gerhardt, Yingyun Cai, Lindsay Marron, Vladimir Lukin, Michael Holbrook. Scalable, Micro-Neutralization Assay for Assessment of SARS-CoV-2 (COVID-19) Virus-Neutralizing Antibodies in Human Clinical Samples. Viruses. 2021; 13 (5):893.
Chicago/Turabian StyleRichard Bennett; Elena Postnikova; Janie Liang; Robin Gross; Steven Mazur; Saurabh Dixit; Gregory Kocher; Shuiqing Yu; Shalamar Georgia-Clark; Dawn Gerhardt; Yingyun Cai; Lindsay Marron; Vladimir Lukin; Michael Holbrook. 2021. "Scalable, Micro-Neutralization Assay for Assessment of SARS-CoV-2 (COVID-19) Virus-Neutralizing Antibodies in Human Clinical Samples." Viruses 13, no. 5: 893.
The emergence of SARS-CoV-2 variants that threaten the efficacy of existing vaccines and therapeutic antibodies underscores the urgent need for new antibody-based tools that potently neutralize variants by targeting multiple sites of the spike protein. We isolated 216 monoclonal antibodies targeting SARS-CoV-2 from plasmablasts and memory B cells of COVID-19 patients. The three most potent antibodies targeted distinct regions of the RBD, and all three neutralized the SARS-CoV-2 variants B.1.1.7 and B.1.351. The crystal structure of the most potent antibody, CV503, revealed that it binds to the ridge region of SARS-CoV-2 RBD, competes with the ACE2 receptor, and has limited contact with key variant residues K417, E484 and N501. We designed bispecific antibodies by combining non-overlapping specificities and identified five ultrapotent bispecific antibodies that inhibit authentic SARS-CoV-2 infection at concentrations of 100-fold more potent than a cocktail of its parent monoclonals in vitro and prevented clinical disease in a hamster model at a 2.5 mg/kg dose. Notably, six of nine bispecific antibodies neutralized B.1.1.7, B.1.351 and the wild-type virus with comparable potency, despite partial or complete loss of activity of at least one parent monoclonal antibody against B.1.351. Furthermore, a bispecific antibody that neutralized B.1.351 protected against SARS-CoV-2 expressing the crucial E484K mutation in the hamster model. Thus, bispecific antibodies represent a promising next-generation countermeasure against SARS-CoV-2 variants of concern.
Hyeseon Cho; Kristina Kay Gonzales-Wartz; Deli Huang; Meng Yuan; Mary Peterson; Janie Liang; Nathan Beutler; Jonathan L. Torres; Yu Cong; Elena Postnikova; Sandhya Bangaru; Chloe Adrienna Talana; Wei Shi; Eun Sung Yang; Yi Zhang; Kwanyee Leung; Lingshu Wang; Linghang Peng; Jeff Skinner; Shanping Li; Nicholas C. Wu; Hejun Liu; Cherrelle Dacon; Thomas Moyer; Melanie Cohen; Ming Zhao; F. Eun-Hyung Lee; Rona S. Weinberg; Iyadh Douagi; Robin Gross; Connie Schmaljohn; Amarendra Pegu; John R. Mascola; Michael Holbrook; David Nemazee; Thomas F. Rogers; Andrew B. Ward; Ian A. Wilson; Peter D. Crompton; Joshua Tan. Ultrapotent bispecific antibodies neutralize emerging SARS-CoV-2 variants. 2021, 1 .
AMA StyleHyeseon Cho, Kristina Kay Gonzales-Wartz, Deli Huang, Meng Yuan, Mary Peterson, Janie Liang, Nathan Beutler, Jonathan L. Torres, Yu Cong, Elena Postnikova, Sandhya Bangaru, Chloe Adrienna Talana, Wei Shi, Eun Sung Yang, Yi Zhang, Kwanyee Leung, Lingshu Wang, Linghang Peng, Jeff Skinner, Shanping Li, Nicholas C. Wu, Hejun Liu, Cherrelle Dacon, Thomas Moyer, Melanie Cohen, Ming Zhao, F. Eun-Hyung Lee, Rona S. Weinberg, Iyadh Douagi, Robin Gross, Connie Schmaljohn, Amarendra Pegu, John R. Mascola, Michael Holbrook, David Nemazee, Thomas F. Rogers, Andrew B. Ward, Ian A. Wilson, Peter D. Crompton, Joshua Tan. Ultrapotent bispecific antibodies neutralize emerging SARS-CoV-2 variants. . 2021; ():1.
Chicago/Turabian StyleHyeseon Cho; Kristina Kay Gonzales-Wartz; Deli Huang; Meng Yuan; Mary Peterson; Janie Liang; Nathan Beutler; Jonathan L. Torres; Yu Cong; Elena Postnikova; Sandhya Bangaru; Chloe Adrienna Talana; Wei Shi; Eun Sung Yang; Yi Zhang; Kwanyee Leung; Lingshu Wang; Linghang Peng; Jeff Skinner; Shanping Li; Nicholas C. Wu; Hejun Liu; Cherrelle Dacon; Thomas Moyer; Melanie Cohen; Ming Zhao; F. Eun-Hyung Lee; Rona S. Weinberg; Iyadh Douagi; Robin Gross; Connie Schmaljohn; Amarendra Pegu; John R. Mascola; Michael Holbrook; David Nemazee; Thomas F. Rogers; Andrew B. Ward; Ian A. Wilson; Peter D. Crompton; Joshua Tan. 2021. "Ultrapotent bispecific antibodies neutralize emerging SARS-CoV-2 variants." , no. : 1.
Outbreaks of Ebola ebolavirus (EBOV) have been associated with high morbidity and mortality. Milestones have been reached recently in the management of EBOV disease (EVD) with licensure of an EBOV vaccine and two monoclonal antibody therapies. However, neither vaccines nor therapies are available for other disease-causing filoviruses. In preparation for such outbreaks, and for more facile and cost-effective management of EVD, we seek a cocktail containing orally available and room temperature stable drugs with strong activity against multiple filoviruses. We previously showed that (bepridil + sertraline) and (sertraline + toremifene) synergistically suppress EBOV in cell cultures. Here, we describe steps towards testing these combinations in a mouse model of EVD. We identified a vehicle suitable for oral delivery of the component drugs and determined that, thus formulated the drugs are equally active against EBOV as preparations in DMSO, and they maintain activity upon storage in solution for up to seven days. Pharmacokinetic (PK) studies indicated that the drugs in the oral delivery vehicle are well tolerated in mice at the highest doses tested. Collectively the data support advancement of these combinations to tests for synergy in a mouse model of EVD. Moreover, mathematical modeling based on human oral PK projects that the combinations would be more active in humans than their component single drugs.
Courtney Finch; Julie Dyall; Shuang Xu; Elizabeth Nelson; Elena Postnikova; Janie Liang; Huanying Zhou; Lisa DeWald; Craig Thomas; Amy Wang; Xin Xu; Emma Hughes; Patrick Morris; Jon Mirsalis; Linh Nguyen; Maria Arolfo; Bryan Koci; Michael Holbrook; Lisa Hensley; Peter Jahrling; Connie Schmaljohn; Lisa Johansen; Gene Olinger; Joshua Schiffer; Judith White. Formulation, Stability, Pharmacokinetic, and Modeling Studies for Tests of Synergistic Combinations of Orally Available Approved Drugs against Ebola Virus In Vivo. Microorganisms 2021, 9, 566 .
AMA StyleCourtney Finch, Julie Dyall, Shuang Xu, Elizabeth Nelson, Elena Postnikova, Janie Liang, Huanying Zhou, Lisa DeWald, Craig Thomas, Amy Wang, Xin Xu, Emma Hughes, Patrick Morris, Jon Mirsalis, Linh Nguyen, Maria Arolfo, Bryan Koci, Michael Holbrook, Lisa Hensley, Peter Jahrling, Connie Schmaljohn, Lisa Johansen, Gene Olinger, Joshua Schiffer, Judith White. Formulation, Stability, Pharmacokinetic, and Modeling Studies for Tests of Synergistic Combinations of Orally Available Approved Drugs against Ebola Virus In Vivo. Microorganisms. 2021; 9 (3):566.
Chicago/Turabian StyleCourtney Finch; Julie Dyall; Shuang Xu; Elizabeth Nelson; Elena Postnikova; Janie Liang; Huanying Zhou; Lisa DeWald; Craig Thomas; Amy Wang; Xin Xu; Emma Hughes; Patrick Morris; Jon Mirsalis; Linh Nguyen; Maria Arolfo; Bryan Koci; Michael Holbrook; Lisa Hensley; Peter Jahrling; Connie Schmaljohn; Lisa Johansen; Gene Olinger; Joshua Schiffer; Judith White. 2021. "Formulation, Stability, Pharmacokinetic, and Modeling Studies for Tests of Synergistic Combinations of Orally Available Approved Drugs against Ebola Virus In Vivo." Microorganisms 9, no. 3: 566.
Background Characterizing the kinetics of the antibody response to SARS□CoV□2 is of critical importance to developing strategies that may mitigate the public health burden of COVID-19. We sought to determine how circulating antibody levels change over time following natural infection. Methods/Materials We conducted a prospective, longitudinal analysis of COVID-19 convalescent plasma (CCP) donors at multiple time points over a 9-month period. At each study visit, subjects either donated plasma or only had study samples drawn. In all cases, anti-SARS-CoV-2 donor testing was performed using semi-quantitative chemiluminescent immunoassays (ChLIA) targeting subunit 1 (S1) of the SARS-CoV-2 spike (S) protein, and an in-house fluorescence reduction neutralization assay (FRNA). Results From April to November 2020 we enrolled 202 donors, mean age 47.3 ±14.7 years, 55% female, 75% Caucasian. Most donors reported a mild clinical course (91%, n=171) without hospitalization. One hundred and five (105) (52%) donors presented for repeat visits with a median 42 (12-163) days between visits. The final visit occurred at a median 160 (53-273) days post-symptom resolution. Total anti-SARS-CoV-2 antibodies (Ab), SARS-CoV-2 specific IgG and neutralizing antibodies were detected in 97.5%, 91.1%, and 74% of donors respectively at initial presentation. Neutralizing Ab titers based on FRNA50 were positively associated with mean IgG levels (p = 50 decreased in 44.4% and remained unchanged in 33.3% of repeat donors. A weak negative correlation was observed between total Ab levels and number of days post-symptom recovery (r = 0.09). Conclusion Anti-SARS-CoV-2 antibodies were identified in 97% of convalescent donors at initial presentation. In a cohort that largely did not require hospitalization. IgG and neutralizing antibodies were positively correlated with age, BMI and clinical severity, and persisted for up to 9 months post-recovery from natural infection. On repeat presentation, IgG anti-SARS-CoV-2 levels decreased in 56% of repeat donors. Overall, these data suggest that CP donors possess a wide range of IgG and neutralizing antibody levels that are proportionally distributed across demographics, with the exception of age, BMI and clinical severity.
Valeria De Giorgi; Kamille A West; Amanda N Henning; Leonard Chen; Michael R Holbrook; Robin Gross; Janie Liang; Elena Postnikova; Joni Trenbeath; Sarah Pogue; Tania Scinto; Harvey J Alter; Cathy Corny Cantilena. Anti-SARS-CoV-2 Serology persistence over time in COVID-19 Convalescent Plasma Donors. 2021, 1 .
AMA StyleValeria De Giorgi, Kamille A West, Amanda N Henning, Leonard Chen, Michael R Holbrook, Robin Gross, Janie Liang, Elena Postnikova, Joni Trenbeath, Sarah Pogue, Tania Scinto, Harvey J Alter, Cathy Corny Cantilena. Anti-SARS-CoV-2 Serology persistence over time in COVID-19 Convalescent Plasma Donors. . 2021; ():1.
Chicago/Turabian StyleValeria De Giorgi; Kamille A West; Amanda N Henning; Leonard Chen; Michael R Holbrook; Robin Gross; Janie Liang; Elena Postnikova; Joni Trenbeath; Sarah Pogue; Tania Scinto; Harvey J Alter; Cathy Corny Cantilena. 2021. "Anti-SARS-CoV-2 Serology persistence over time in COVID-19 Convalescent Plasma Donors." , no. : 1.
As the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic was expanding, it was clear that effective testing for the presence of neutralizing antibodies in the blood of convalescent patients would be critical for development of plasma-based therapeutic approaches. To address the need for a high-quality neutralization assay against SARS-CoV-2, a previously established fluorescence reduction neutralization assay (FRNA) against Middle East respiratory syndrome coronavirus (MERS-CoV) was modified and optimized. The SARS-CoV-2 FRNA provides a quantitative assessment of a large number of infected cells through use of a high-content imaging system. Because of this approach, and the fact that it does not involve subjective interpretation, this assay is more efficient and more accurate than other neutralization assays. In addition, the ability to set robust acceptance criteria for individual plates and specific test wells provided further rigor to this assay. Such agile adaptability avails use with multiple virus variants. By February 2021, the SARS-CoV-2 FRNA had been used to screen over 5,000 samples, including acute and convalescent plasma or serum samples and therapeutic antibody treatments, for SARS-CoV-2 neutralizing titers.
Richard S. Bennett; Elena N. Postnikova; Janie Liang; Robin Gross; Steven Mazur; Saurabh Dixit; Vladimir V. Lukin; Greg Kocher; Shuiqing Yu; Shalamar Georgia-Clark; Dawn Gerhardt; Yingyun Cai; Lindsay Marron; Michael R. Holbrook. Scalable, Micro-Neutralization Assay for Qualitative Assessment of SARS-CoV-2 (COVID-19) Virus-Neutralizing Antibodies in Human Clinical Samples. 2021, 1 .
AMA StyleRichard S. Bennett, Elena N. Postnikova, Janie Liang, Robin Gross, Steven Mazur, Saurabh Dixit, Vladimir V. Lukin, Greg Kocher, Shuiqing Yu, Shalamar Georgia-Clark, Dawn Gerhardt, Yingyun Cai, Lindsay Marron, Michael R. Holbrook. Scalable, Micro-Neutralization Assay for Qualitative Assessment of SARS-CoV-2 (COVID-19) Virus-Neutralizing Antibodies in Human Clinical Samples. . 2021; ():1.
Chicago/Turabian StyleRichard S. Bennett; Elena N. Postnikova; Janie Liang; Robin Gross; Steven Mazur; Saurabh Dixit; Vladimir V. Lukin; Greg Kocher; Shuiqing Yu; Shalamar Georgia-Clark; Dawn Gerhardt; Yingyun Cai; Lindsay Marron; Michael R. Holbrook. 2021. "Scalable, Micro-Neutralization Assay for Qualitative Assessment of SARS-CoV-2 (COVID-19) Virus-Neutralizing Antibodies in Human Clinical Samples." , no. : 1.
Outbreaks of Ebola ebolavirus (EBOV) have been associated with high morbidity and mortality. Milestones have been reached recently in the management of EBOV disease (EVD) with licensure of an EBOV vaccine and two monoclonal antibody therapies. However, neither vaccines nor therapies are available for other disease-causing filoviruses. In preparation for such outbreaks, and for more facile and cost-effective management of EVD, we seek a cocktail containing orally available and room temperature stable drugs with strong activity against multiple filoviruses. We previously showed that (bepridil + sertraline) and (sertraline + toremifene) synergistically suppress EBOV in cell cultures. Here we describe steps towards testing these combinations in a mouse model of EVD. We identified a vehicle suitable for oral delivery of the component drugs and determined that, thus formulated the drugs are equally active against EBOV as preparations in DMSO, and they maintain activity upon storage in solution for up to seven days. Pharmacokinetic (PK) studies indicated that the drugs in the oral delivery vehicle are well tolerated in mice at the highest doses tested. Collectively the data support advancement of these combinations to tests for synergy in a mouse model of EVD. Moreover, mathematical modeling based on human oral PK projects that the combinations would be more active in humans than their component single drugs.
Courtney L. Finch; Julie Dyall; Shuang Xu; Elizabeth A. Nelson; Elena Postnikova; Janie Y. Liang; Huanying Zhou; Lisa Evans DeWald; Craig J. Thomas; Amy Wang; Xin Xu; Emma Hughes; Patrick J. Morris; Jon C. Mirsalis; Linh H. Nguyen; Maria P. Arolfo; Bryan Koci; Michael R. Holbrook; Lisa E. Hensley; Peter B. Jahrling; Connie Schmaljohn; Lisa M. Johansen; Gene G. Olinger; Joshua T. Schiffer; Judith White. Formulation, Stability, Pharmacokinetic, and Modeling Studies for Tests of Synergistic Combinations of Orally Available Approved Drugs Against Ebola Virus In Vivo. 2021, 1 .
AMA StyleCourtney L. Finch, Julie Dyall, Shuang Xu, Elizabeth A. Nelson, Elena Postnikova, Janie Y. Liang, Huanying Zhou, Lisa Evans DeWald, Craig J. Thomas, Amy Wang, Xin Xu, Emma Hughes, Patrick J. Morris, Jon C. Mirsalis, Linh H. Nguyen, Maria P. Arolfo, Bryan Koci, Michael R. Holbrook, Lisa E. Hensley, Peter B. Jahrling, Connie Schmaljohn, Lisa M. Johansen, Gene G. Olinger, Joshua T. Schiffer, Judith White. Formulation, Stability, Pharmacokinetic, and Modeling Studies for Tests of Synergistic Combinations of Orally Available Approved Drugs Against Ebola Virus In Vivo. . 2021; ():1.
Chicago/Turabian StyleCourtney L. Finch; Julie Dyall; Shuang Xu; Elizabeth A. Nelson; Elena Postnikova; Janie Y. Liang; Huanying Zhou; Lisa Evans DeWald; Craig J. Thomas; Amy Wang; Xin Xu; Emma Hughes; Patrick J. Morris; Jon C. Mirsalis; Linh H. Nguyen; Maria P. Arolfo; Bryan Koci; Michael R. Holbrook; Lisa E. Hensley; Peter B. Jahrling; Connie Schmaljohn; Lisa M. Johansen; Gene G. Olinger; Joshua T. Schiffer; Judith White. 2021. "Formulation, Stability, Pharmacokinetic, and Modeling Studies for Tests of Synergistic Combinations of Orally Available Approved Drugs Against Ebola Virus In Vivo." , no. : 1.
The SARS-CoV-2 pandemic has made it clear that we have a desperate need for antivirals. We present work that the mammalian SKI complex is a broad-spectrum, host-directed, antiviral drug target. Yeast suppressor screening was utilized to find a functional genetic interaction between proteins from influenza A virus (IAV) and Middle East respiratory syndrome coronavirus (MERS-CoV) with eukaryotic proteins that may be potential host factors involved in replication. This screening identified the SKI complex as a potential host factor for both viruses. In mammalian systems siRNA-mediated knockdown of SKI genes inhibited replication of IAV and MERS-CoV. In silico modeling and database screening identified a binding pocket on the SKI complex and compounds predicted to bind. Experimental assays of those compounds identified three chemical structures that were antiviral against IAV and MERS-CoV along with the filoviruses Ebola and Marburg and two further coronaviruses, SARS-CoV and SARS-CoV-2. The mechanism of antiviral activity is through inhibition of viral RNA production. This work defines the mammalian SKI complex as a broad-spectrum antiviral drug target and identifies lead compounds for further development.
Stuart Weston; Lauren Baracco; Chloe Keller; Krystal Matthews; Marisa E. McGrath; James Logue; Janie Liang; Julie Dyall; Michael R. Holbrook; Lisa E. Hensley; Peter B. Jahrling; Wenbo Yu; Alexander D. MacKerell Jr; Matthew B. Frieman. The SKI complex is a broad-spectrum, host-directed antiviral drug target for coronaviruses, influenza, and filoviruses. Proceedings of the National Academy of Sciences 2020, 117, 30687 -30698.
AMA StyleStuart Weston, Lauren Baracco, Chloe Keller, Krystal Matthews, Marisa E. McGrath, James Logue, Janie Liang, Julie Dyall, Michael R. Holbrook, Lisa E. Hensley, Peter B. Jahrling, Wenbo Yu, Alexander D. MacKerell Jr, Matthew B. Frieman. The SKI complex is a broad-spectrum, host-directed antiviral drug target for coronaviruses, influenza, and filoviruses. Proceedings of the National Academy of Sciences. 2020; 117 (48):30687-30698.
Chicago/Turabian StyleStuart Weston; Lauren Baracco; Chloe Keller; Krystal Matthews; Marisa E. McGrath; James Logue; Janie Liang; Julie Dyall; Michael R. Holbrook; Lisa E. Hensley; Peter B. Jahrling; Wenbo Yu; Alexander D. MacKerell Jr; Matthew B. Frieman. 2020. "The SKI complex is a broad-spectrum, host-directed antiviral drug target for coronaviruses, influenza, and filoviruses." Proceedings of the National Academy of Sciences 117, no. 48: 30687-30698.
Background: SARS-CoV-2-specific antibodies may protect from reinfection and disease, providing the rationale for administration of plasma containing SARS-CoV-2 neutralizing antibodies (nAb) as a treatment for COVID-19. The clinical factors and laboratory assays to streamline plasma donor selection, and the durability of nAb responses, are incompletely understood. Methods: Adults with virologically-documented SARS-CoV-2 infection in a convalescent plasma donor screening program were tested for serum IgG to SARS-CoV-2 spike protein S1 domain, nucleoprotein (NP), and for nAb. Results: Amongst 250 consecutive persons studied a median of 67 days since symptom onset, 243/250 (97%) were seropositive on one or more assays. Sixty percent of donors had nAb titers ≥1:80. Correlates of higher nAb titer included older age (adjusted OR [AOR] 1.03/year of age, 95% CI 1.00-1.06), male sex (AOR 2.08, 95% CI 1.13-3.82), fever during acute illness (AOR 2.73, 95% CI 1.25-5.97), and disease severity represented by hospitalization (AOR 6.59, 95% CI 1.32-32.96). Receiver operating characteristic (ROC) analyses of anti-S1 and anti-NP antibody results yielded cutoffs that corresponded well with nAb titers, with the anti-S1 assay being slightly more predictive. NAb titers declined in 37 of 41 paired specimens collected a median of 98 days (range, 77-120) apart (P
Jim Boonyaratanakornkit; Chihiro Morishima; Stacy Selke; Danniel Zamora; Sarah McGuffin; Adrienne E Shapiro; Victoria L Campbell; Christopher L McClurkan; Lichen Jing; Robin Gross; Janie Liang; Elena Postnikova; Steven Mazur; Anu Chaudhary; Marie K Das; Susan L Fink; Andrew Bryan; Alex L Greninger; Keith R Jerome; Michael R Holbrook; Terry B Gernsheimer; Mark H Wener; Anna Wald; David M Koelle. Clinical, laboratory, and temporal predictors of neutralizing antibodies to SARS-CoV-2 after COVID-19. 2020, 1 .
AMA StyleJim Boonyaratanakornkit, Chihiro Morishima, Stacy Selke, Danniel Zamora, Sarah McGuffin, Adrienne E Shapiro, Victoria L Campbell, Christopher L McClurkan, Lichen Jing, Robin Gross, Janie Liang, Elena Postnikova, Steven Mazur, Anu Chaudhary, Marie K Das, Susan L Fink, Andrew Bryan, Alex L Greninger, Keith R Jerome, Michael R Holbrook, Terry B Gernsheimer, Mark H Wener, Anna Wald, David M Koelle. Clinical, laboratory, and temporal predictors of neutralizing antibodies to SARS-CoV-2 after COVID-19. . 2020; ():1.
Chicago/Turabian StyleJim Boonyaratanakornkit; Chihiro Morishima; Stacy Selke; Danniel Zamora; Sarah McGuffin; Adrienne E Shapiro; Victoria L Campbell; Christopher L McClurkan; Lichen Jing; Robin Gross; Janie Liang; Elena Postnikova; Steven Mazur; Anu Chaudhary; Marie K Das; Susan L Fink; Andrew Bryan; Alex L Greninger; Keith R Jerome; Michael R Holbrook; Terry B Gernsheimer; Mark H Wener; Anna Wald; David M Koelle. 2020. "Clinical, laboratory, and temporal predictors of neutralizing antibodies to SARS-CoV-2 after COVID-19." , no. : 1.
We have recently identified three molecules (tilorone, quinacrine and pyronaridine tetraphosphate) which all demonstrated efficacy in the mouse model of infection with mouse-adapted Ebola virus (EBOV) model of disease and had similar in vitro inhibition of an Ebola pseudovirus (VSV-EBOV-GP), suggesting they interfere with viral entry. Using a machine learning model to predict lysosomotropism these compounds were evaluated for their ability to inhibit via a lysosomotropic mechanism in vitro. We now demonstrate in vitro that pyronaridine tetraphosphate is an inhibitor of Lysotracker accumulation in lysosomes (IC50 = 0.56 μM). Further, we evaluated synergy between pyronaridine and artesunate (Pyramax®), which are used in combination to treat malaria. Artesunate was not found to have lysosomotropic activity in vitro and the combination effect on EBOV inhibition was shown to be additive. Pyramax® may represent a unique example of the repurposing of a combination product for another disease.
Thomas R. Lane; Julie Dyall; Luke Mercer; Caleb Goodin; Daniel H. Foil; Huanying Zhou; Elena Postnikova; Janie Y. Liang; Michael R. Holbrook; Peter B. Madrid; Sean Ekins. Repurposing Pyramax® for the Treatment of Ebola Virus Disease: Additivity of the Lysosomotropic Pyronaridine and Non-Lysosomotropic Artesunate. 2020, 1 .
AMA StyleThomas R. Lane, Julie Dyall, Luke Mercer, Caleb Goodin, Daniel H. Foil, Huanying Zhou, Elena Postnikova, Janie Y. Liang, Michael R. Holbrook, Peter B. Madrid, Sean Ekins. Repurposing Pyramax® for the Treatment of Ebola Virus Disease: Additivity of the Lysosomotropic Pyronaridine and Non-Lysosomotropic Artesunate. . 2020; ():1.
Chicago/Turabian StyleThomas R. Lane; Julie Dyall; Luke Mercer; Caleb Goodin; Daniel H. Foil; Huanying Zhou; Elena Postnikova; Janie Y. Liang; Michael R. Holbrook; Peter B. Madrid; Sean Ekins. 2020. "Repurposing Pyramax® for the Treatment of Ebola Virus Disease: Additivity of the Lysosomotropic Pyronaridine and Non-Lysosomotropic Artesunate." , no. : 1.
During the Ebola virus disease (EVD) epidemic in Western Africa (2013‒2016), antimalarial treatment was administered to EVD patients due to the high coexisting malaria burden in accordance with World Health Organization guidelines. In an Ebola treatment center in Liberia, EVD patients receiving the combination antimalarial artesunate-amodiaquine had a lower risk of death compared to those treated with artemether-lumefantrine. As artemether and artesunate are derivatives of artemisinin, the beneficial anti-Ebola virus (EBOV) effect observed could possibly be attributed to the change from lumefantrine to amodiaquine. Amodiaquine is a widely used antimalarial in the countries that experience outbreaks of EVD and, therefore, holds promise as an approved drug that could be repurposed for treating EBOV infections. We investigated the potential anti-EBOV effect of amodiaquine in a well-characterized nonhuman primate model of EVD. Using a similar 3-day antimalarial dosing strategy as for human patients, plasma concentrations of amodiaquine in healthy animals were similar to those found in humans. However, the treatment regimen did not result in a survival benefit or decrease of disease signs in EBOV-infected animals. While amodiaquine on its own failed to demonstrate efficacy, we cannot exclude potential therapeutic value of amodiaquine when used in combination with artesunate or another antiviral.
Lisa Evans Dewald; Joshua Johnson; Dawn M. Gerhardt; Lisa M. Torzewski; Elena Postnikova; Anna N. Honko; Krisztina Janosko; Louis Huzella; William E. Dowling; Ann E. Eakin; Blaire L. Osborn; Janet Gahagen; Liang Tang; Carol E. Green; Jon C. Mirsalis; Michael R. Holbrook; Peter B. Jahrling; Julie Dyall; Lisa Hensley. In Vivo Activity of Amodiaquine against Ebola Virus Infection. Scientific Reports 2019, 9, 20199 -12.
AMA StyleLisa Evans Dewald, Joshua Johnson, Dawn M. Gerhardt, Lisa M. Torzewski, Elena Postnikova, Anna N. Honko, Krisztina Janosko, Louis Huzella, William E. Dowling, Ann E. Eakin, Blaire L. Osborn, Janet Gahagen, Liang Tang, Carol E. Green, Jon C. Mirsalis, Michael R. Holbrook, Peter B. Jahrling, Julie Dyall, Lisa Hensley. In Vivo Activity of Amodiaquine against Ebola Virus Infection. Scientific Reports. 2019; 9 (1):20199-12.
Chicago/Turabian StyleLisa Evans Dewald; Joshua Johnson; Dawn M. Gerhardt; Lisa M. Torzewski; Elena Postnikova; Anna N. Honko; Krisztina Janosko; Louis Huzella; William E. Dowling; Ann E. Eakin; Blaire L. Osborn; Janet Gahagen; Liang Tang; Carol E. Green; Jon C. Mirsalis; Michael R. Holbrook; Peter B. Jahrling; Julie Dyall; Lisa Hensley. 2019. "In Vivo Activity of Amodiaquine against Ebola Virus Infection." Scientific Reports 9, no. 1: 20199-12.
Nipah virus (NiV) is an emerging virus associated with outbreaks of acute respiratory disease and encephalitis. To develop a neurological model for NiV infection, we exposed 6 adult African green monkeys to a large-particle (approximately 12 μm) aerosol containing NiV (Malaysian isolate). Brain magnetic resonance images were obtained at baseline, every 3 days after exposure for 2 weeks, and then weekly until week 8 after exposure. Four of six animals showed abnormalities reminiscent of human disease in brain magnetic resonance images. Abnormalities ranged from cytotoxic edema to vasogenic edema. The majority of lesions were small infarcts, and a few showed inflammatory or encephalitic changes. Resolution or decreased size in some lesions resembled findings reported in patients with NiV infection. Histological lesions in the brain included multifocal areas of encephalomalacia, corresponding to known ischemic foci. In other regions of the brain there was evidence of vasculitis, with perivascular infiltrates of inflammatory cells and rare intravascular fibrin thrombi. This animal model will help us better understand the acute neurological features of NiV infection and develop therapeutic approaches for managing disease caused by NiV infection.
Ji Hyun Lee; Dima A Hammoud; Yu Cong; Louis M Huzella; Marcelo A Castro; Jeffrey Solomon; Joseph Laux; Matthew Lackemeyer; J Kyle Bohannon; Oscar Rojas; Russ Byrum; Ricky Adams; Danny Ragland; Marisa St Claire; Vincent Munster; Michael R Holbrook. The Use of Large-Particle Aerosol Exposure to Nipah Virus to Mimic Human Neurological Disease Manifestations in the African Green Monkey. The Journal of Infectious Diseases 2019, 221, S419 -S430.
AMA StyleJi Hyun Lee, Dima A Hammoud, Yu Cong, Louis M Huzella, Marcelo A Castro, Jeffrey Solomon, Joseph Laux, Matthew Lackemeyer, J Kyle Bohannon, Oscar Rojas, Russ Byrum, Ricky Adams, Danny Ragland, Marisa St Claire, Vincent Munster, Michael R Holbrook. The Use of Large-Particle Aerosol Exposure to Nipah Virus to Mimic Human Neurological Disease Manifestations in the African Green Monkey. The Journal of Infectious Diseases. 2019; 221 (Supplement):S419-S430.
Chicago/Turabian StyleJi Hyun Lee; Dima A Hammoud; Yu Cong; Louis M Huzella; Marcelo A Castro; Jeffrey Solomon; Joseph Laux; Matthew Lackemeyer; J Kyle Bohannon; Oscar Rojas; Russ Byrum; Ricky Adams; Danny Ragland; Marisa St Claire; Vincent Munster; Michael R Holbrook. 2019. "The Use of Large-Particle Aerosol Exposure to Nipah Virus to Mimic Human Neurological Disease Manifestations in the African Green Monkey." The Journal of Infectious Diseases 221, no. Supplement: S419-S430.
The ability to appropriately mimic human disease is critical for using animal models as a tool for understanding virus pathogenesis. In the case of Nipah virus (NiV), infection of humans appears to occur either through inhalation, contact with or consumption of infected material. In two of these circumstances, respiratory or sinusoidal exposure represents a likely route of infection. In this study, intermediate-size aerosol particles (~7 μm) of NiV-Malaysia were used to mimic potential routes of exposure by focusing viral deposition in the upper respiratory tract. Our previous report showed this route of exposure extended the disease course and a single animal survived the infection. Here, analysis of the peripheral immune response found minimal evidence of systemic inflammation and depletion of B cells during acute disease. However, the animal that survived infection developed an early IgM response with rapid development of neutralizing antibodies that likely afforded protection. The increase in NiV-specific antibodies correlated with an expansion of the B cell population in the survivor. Cell-mediated immunity was not clearly apparent in animals that succumbed during the acute phase of disease. However, CD4+ and CD8+ effector memory cells increased in the survivor with correlating increases in cytokines and chemokines associated with cell-mediated immunity. Interestingly, kinetic changes of the CD4+ and CD8bright T cell populations over the course of acute disease were opposite from animals that succumbed to infection. In addition, increases in NK cells and basophils during convalescence of the surviving animal were also evident, with viral antigen found in NK cells. These data suggest that a systemic inflammatory response and “cytokine storm” are not major contributors to NiV-Malaysia pathogenesis in the AGM model using this exposure route. Further, these data demonstrate that regulation of cell-mediated immunity, in addition to rapid production of NiV specific antibodies, may be critical for surviving NiV infection. Nipah virus (NiV) infection in Malaysia, Bangladesh and India has been correlated with severe respiratory and neurological disease that led to death in over 50% of known cases. In this study, we used a nonhuman primate model for NiV infection to evaluate the peripheral immune response to virus infection in an effort to identify aspects of the immune response that may be important for survival. An aerosol exposure that targeted virus deposition in sinuses and upper respiratory tract was used in an effort to mimic a probable human exposure route. Following exposure, five of six animals included in the study succumbed to the infection. The survivor developed a virus-specific antibody response and showed clear evidence of cell-mediated immunity. Interestingly, the rate of change in CD4+ and CD8bright T cell populations in the survivor over the course of the acute disease, were the reverse of animals that succumbed to infection. These data suggest that rapid development of virus-specific adaptive immunity is critical for survival of NiV infection.
Abigail Lara; Yu Cong; Peter B. Jahrling; Mark Mednikov; Elena Postnikova; Shuiqing Yu; Vincent Munster; Michael R. Holbrook. Peripheral immune response in the African green monkey model following Nipah-Malaysia virus exposure by intermediate-size particle aerosol. PLOS Neglected Tropical Diseases 2019, 13, e0007454 .
AMA StyleAbigail Lara, Yu Cong, Peter B. Jahrling, Mark Mednikov, Elena Postnikova, Shuiqing Yu, Vincent Munster, Michael R. Holbrook. Peripheral immune response in the African green monkey model following Nipah-Malaysia virus exposure by intermediate-size particle aerosol. PLOS Neglected Tropical Diseases. 2019; 13 (6):e0007454.
Chicago/Turabian StyleAbigail Lara; Yu Cong; Peter B. Jahrling; Mark Mednikov; Elena Postnikova; Shuiqing Yu; Vincent Munster; Michael R. Holbrook. 2019. "Peripheral immune response in the African green monkey model following Nipah-Malaysia virus exposure by intermediate-size particle aerosol." PLOS Neglected Tropical Diseases 13, no. 6: e0007454.
Neurotropic viruses can cause rapidly progressing and potentially devastating disease. In this chapter, predominant enveloped RNA viruses that can induce neurological disease are discussed. Some of these viruses, such as alphaviruses and flaviviruses, are transmitted by the bite of an arthropod. Others are transmitted through direct contact with infected animals or their excreta. While several of these viruses have been explored as potential bioweapons, the potential for such an application is limited by the technical ability to generate sufficient virus stocks and the poor transmissibility of these viruses. Also discussed are existing or potential medical countermeasures for the treatment or prevention of infection.
Michael R. Holbrook. Neurotropic Viruses. Defense Against Biological Attacks 2019, 1 -20.
AMA StyleMichael R. Holbrook. Neurotropic Viruses. Defense Against Biological Attacks. 2019; ():1-20.
Chicago/Turabian StyleMichael R. Holbrook. 2019. "Neurotropic Viruses." Defense Against Biological Attacks , no. : 1-20.
Nipah virus (NiV) infection can lead to severe respiratory or neurological disease in humans. Transmission of NiV has been shown to occur through contact with virus contaminated fomites or consumption of contaminated food. Previous results using the African green monkey (AGM) model of NiV infection identified aspects of infection that, while similar to humans, don’t fully recapitulate disease. Previous studies also demonstrate near uniform lethality that is not consistent with human NiV infection. In these studies, aerosol exposure using an intermediate particle size (7μm) was used to mimic potential human exposure by facilitating virus deposition in the upper respiratory tract. Computed tomography evaluation found some animals developed pulmonary parenchymal disease including consolidations, ground-glass opacities, and reactive adenopathy. Despite the lack of neurological signs, magnetic resonance imaging identified distinct brain lesions in three animals, similar to those previously reported in NiV-infected patients. Immunological characterization of tissues collected at necropsy suggested a local pulmonary inflammatory response with increased levels of macrophages in the lung, but a limited neurologic response. These data provide the first clear evidence of neurological involvement in the AGM that recapitulates human disease. With the development of a disease model that is more representative of human disease, these data suggest that NiV infection in the AGM may be appropriate for evaluating therapeutic countermeasures directed at virus-induced neuropathogenesis. The development of effective therapeutic approaches to the treatment of human diseases requires an understanding of the disease process induced by an infectious agent. Historically the development of medical countermeasures for highly pathogenic viruses required the use of a uniformly lethal animal model. While this approach is useful in some regards, it frequently does not provide a true indication of the disease process. In the work presented here, the approach was to use a virus exposure method that mimicked a potential route of human exposure and used a dose that might be more representative of one a human would receive. Using this method and advanced medical imaging techniques, we were able to demonstrate an extended disease course with mixed respiratory and neurological disease like that seen in humans. This study also found that the response to infection in the lungs was inflammatory and that the disease in the brain was limited despite clear evidence of lesions. These data support the development of animal models that mimic human disease and allow for the identification of potential therapeutic approaches that target the disease process rather than only the virus.
Dima Hammoud; Margaret R. Lentz; Abigail Lara; Jordan K. Bohannon; Irwin Feuerstein; Louis Huzella; Peter B. Jahrling; Matthew Lackemeyer; Joseph Laux; Oscar Rojas; Philip Sayre; Jeffrey Solomon; Yu Cong; Vincent Munster; Michael R. Holbrook. Aerosol exposure to intermediate size Nipah virus particles induces neurological disease in African green monkeys. PLOS Neglected Tropical Diseases 2018, 12, e0006978 .
AMA StyleDima Hammoud, Margaret R. Lentz, Abigail Lara, Jordan K. Bohannon, Irwin Feuerstein, Louis Huzella, Peter B. Jahrling, Matthew Lackemeyer, Joseph Laux, Oscar Rojas, Philip Sayre, Jeffrey Solomon, Yu Cong, Vincent Munster, Michael R. Holbrook. Aerosol exposure to intermediate size Nipah virus particles induces neurological disease in African green monkeys. PLOS Neglected Tropical Diseases. 2018; 12 (11):e0006978.
Chicago/Turabian StyleDima Hammoud; Margaret R. Lentz; Abigail Lara; Jordan K. Bohannon; Irwin Feuerstein; Louis Huzella; Peter B. Jahrling; Matthew Lackemeyer; Joseph Laux; Oscar Rojas; Philip Sayre; Jeffrey Solomon; Yu Cong; Vincent Munster; Michael R. Holbrook. 2018. "Aerosol exposure to intermediate size Nipah virus particles induces neurological disease in African green monkeys." PLOS Neglected Tropical Diseases 12, no. 11: e0006978.
Lisa Evans Dewald; Julie Dyall; Jennifer M Sword; Lisa Torzewski; Huanying Zhou; Elena Postnikova; Erin Kollins; Isis Alexander; Robin Gross; Yu Cong; Dawn M Gerhardt; Reed F Johnson; Gene G Olinger; Michael R Holbrook; Lisa E Hensley; Peter B Jahrling. The Calcium Channel Blocker Bepridil Demonstrates Efficacy in the Murine Model of Marburg Virus Disease. The Journal of Infectious Diseases 2018, 218, S588 -S591.
AMA StyleLisa Evans Dewald, Julie Dyall, Jennifer M Sword, Lisa Torzewski, Huanying Zhou, Elena Postnikova, Erin Kollins, Isis Alexander, Robin Gross, Yu Cong, Dawn M Gerhardt, Reed F Johnson, Gene G Olinger, Michael R Holbrook, Lisa E Hensley, Peter B Jahrling. The Calcium Channel Blocker Bepridil Demonstrates Efficacy in the Murine Model of Marburg Virus Disease. The Journal of Infectious Diseases. 2018; 218 (suppl_5):S588-S591.
Chicago/Turabian StyleLisa Evans Dewald; Julie Dyall; Jennifer M Sword; Lisa Torzewski; Huanying Zhou; Elena Postnikova; Erin Kollins; Isis Alexander; Robin Gross; Yu Cong; Dawn M Gerhardt; Reed F Johnson; Gene G Olinger; Michael R Holbrook; Lisa E Hensley; Peter B Jahrling. 2018. "The Calcium Channel Blocker Bepridil Demonstrates Efficacy in the Murine Model of Marburg Virus Disease." The Journal of Infectious Diseases 218, no. suppl_5: S588-S591.
A need to develop therapeutics to treat Ebola virus disease patients in remote and resource-challenged settings remains in the wake of the 2013–2016 epidemic in West Africa. Toward this goal, we screened drugs under consideration as treatment options and other drugs of interest, most being small molecules approved by the Food and Drug Administration. Drugs demonstrating in vitro antiviral activity were advanced for evaluation in combinations because of advantages often provided by drug cocktails. Drugs were screened for blockade of Ebola virus infection in cultured cells. Twelve drugs were tested in all (78 pair-wise) combinations, and 3 were tested in a subset of combinations. Multiple synergistic drug pairs emerged, with the majority comprising 2 entry inhibitors. For the pairs of entry inhibitors studied, synergy was demonstrated at the level of virus entry into host cells. Highly synergistic pairs included aripiprazole/piperacetazine, sertraline/toremifene, sertraline/bepridil, and amodiaquine/clomiphene. Our study shows the feasibility of identifying pairs of approved drugs that synergistically block Ebola virus infection in cell cultures. We discuss our findings in terms of the theoretic ability of these or alternate combinations to reach therapeutic levels. Future research will assess selected combinations in small-animal models of Ebola virus disease.
Julie Dyall; Elizabeth A Nelson; Lisa Evans DeWald; Rajarshi Guha; Brit J Hart; Huanying Zhou; Elena Postnikova; James Logue; Walter M Vargas; Robin Gross; Julia Michelotti; Nicole Deiuliis; Richard Bennett; Ian Crozier; Michael R Holbrook; Patrick J Morris; Carleen Klumpp-Thomas; Crystal McKnight; Tim Mierzwa; Paul Shinn; Pamela J Glass; Lisa M Johansen; Peter B Jahrling; Lisa E Hensley; Gene Olinger; Craig Thomas; Judith M White. Identification of Combinations of Approved Drugs With Synergistic Activity Against Ebola Virus in Cell Cultures. Journal of Infectious Diseases 2018, 218, S672 -S678.
AMA StyleJulie Dyall, Elizabeth A Nelson, Lisa Evans DeWald, Rajarshi Guha, Brit J Hart, Huanying Zhou, Elena Postnikova, James Logue, Walter M Vargas, Robin Gross, Julia Michelotti, Nicole Deiuliis, Richard Bennett, Ian Crozier, Michael R Holbrook, Patrick J Morris, Carleen Klumpp-Thomas, Crystal McKnight, Tim Mierzwa, Paul Shinn, Pamela J Glass, Lisa M Johansen, Peter B Jahrling, Lisa E Hensley, Gene Olinger, Craig Thomas, Judith M White. Identification of Combinations of Approved Drugs With Synergistic Activity Against Ebola Virus in Cell Cultures. Journal of Infectious Diseases. 2018; 218 (suppl_5):S672-S678.
Chicago/Turabian StyleJulie Dyall; Elizabeth A Nelson; Lisa Evans DeWald; Rajarshi Guha; Brit J Hart; Huanying Zhou; Elena Postnikova; James Logue; Walter M Vargas; Robin Gross; Julia Michelotti; Nicole Deiuliis; Richard Bennett; Ian Crozier; Michael R Holbrook; Patrick J Morris; Carleen Klumpp-Thomas; Crystal McKnight; Tim Mierzwa; Paul Shinn; Pamela J Glass; Lisa M Johansen; Peter B Jahrling; Lisa E Hensley; Gene Olinger; Craig Thomas; Judith M White. 2018. "Identification of Combinations of Approved Drugs With Synergistic Activity Against Ebola Virus in Cell Cultures." Journal of Infectious Diseases 218, no. suppl_5: S672-S678.
Nipah virus (NiV) is a highly pathogenic zoonotic paramyxovirus that can result in severe pulmonary disease and fatal encephalitis in humans and is responsible for outbreaks in Bangladesh, Malaysia, Singapore, India and possibly the Philippines. NiV has a negative-sense RNA genome that contains six genes and serves as a template for production of viral mRNA transcripts. NiV mRNA transcripts are subsequently translated into viral proteins. Traditionally, NiV quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR) assays have relied on using primer sets that amplify a target (N that encodes the nucleocapsid) within the coding region of the viral gene that also amplifies viral mRNA. Here we describe a novel one-step qRT-PCR assay targeting the intergenic region separating the viral F and G proteins, thereby eliminating amplification of the viral mRNA. This assay is more accurate than the traditional qRT-PCR in quantifying concentrations of viral genomic RNA.
Kenneth S. Jensen; Ricky Adams; Richard S. Bennett; John Bernbaum; Peter B. Jahrling; Michael R. Holbrook. Development of a novel real-time polymerase chain reaction assay for the quantitative detection of Nipah virus replicative viral RNA. PLOS ONE 2018, 13, e0199534 .
AMA StyleKenneth S. Jensen, Ricky Adams, Richard S. Bennett, John Bernbaum, Peter B. Jahrling, Michael R. Holbrook. Development of a novel real-time polymerase chain reaction assay for the quantitative detection of Nipah virus replicative viral RNA. PLOS ONE. 2018; 13 (6):e0199534.
Chicago/Turabian StyleKenneth S. Jensen; Ricky Adams; Richard S. Bennett; John Bernbaum; Peter B. Jahrling; Michael R. Holbrook. 2018. "Development of a novel real-time polymerase chain reaction assay for the quantitative detection of Nipah virus replicative viral RNA." PLOS ONE 13, no. 6: e0199534.
Middle East respiratory syndrome coronavirus (MERS-CoV) presents an emerging threat to public health worldwide by causing severe respiratory disease in humans with high virulence and case fatality rate (about 35%) since 2012. Little is known about the pathogenesis and innate antiviral response in primary human monocyte-derived macrophages (MDMs) and dendritic cells (MDDCs) upon MERS-CoV infection. In this study, we assessed MERS-CoV replication as well as induction of inflammatory cytokines and chemokines in MDMs and immature and mature MDDCs. Immature MDDCs and MDMs were permissive for MERS-CoV infection, while mature MDDCs were not, with stimulation of proinflammatory cytokine and chemokine upregulation in MDMs, but not in MDDCs. To further evaluate the antiviral activity of well-defined drugs in primary antigen presenting cells (APCs), three compounds (chloroquine, chlorpromazine and toremifine), each with broad-spectrum antiviral activity in immortalized cell lines, were evaluated in MDMs and MDDCs to determine their antiviral effect on MERS-CoV infection. While chloroquine was not active in these primary cells, chlorpromazine showed strong anti-MERS-CoV activity, but it was associated with high cytotoxicity narrowing the potential window for drug utilization. Unlike in established cells, toremifene had marginal activity when tested in antigen presenting cells, with high apparent cytotoxicity, also limiting its potential as a therapeutic option. These results demonstrate the value of testing drugs in primary cells, in addition to established cell lines, before initiating preclinical or clinical studies for MERS treatment and the importance of carefully assessing cytotoxicity in drug screen assays. Furthermore, these studies also highlight the role of APCs in stimulating a robust protective immune response to MERS-CoV infection.
Yu Cong; Brit J. Hart; Robin Gross; Huanying Zhou; Matthew Frieman; Laura Bollinger; Jiro Wada; Lisa Hensley; Peter B. Jahrling; Julie Dyall; Michael R. Holbrook. MERS-CoV pathogenesis and antiviral efficacy of licensed drugs in human monocyte-derived antigen-presenting cells. PLOS ONE 2018, 13, e0194868 .
AMA StyleYu Cong, Brit J. Hart, Robin Gross, Huanying Zhou, Matthew Frieman, Laura Bollinger, Jiro Wada, Lisa Hensley, Peter B. Jahrling, Julie Dyall, Michael R. Holbrook. MERS-CoV pathogenesis and antiviral efficacy of licensed drugs in human monocyte-derived antigen-presenting cells. PLOS ONE. 2018; 13 (3):e0194868.
Chicago/Turabian StyleYu Cong; Brit J. Hart; Robin Gross; Huanying Zhou; Matthew Frieman; Laura Bollinger; Jiro Wada; Lisa Hensley; Peter B. Jahrling; Julie Dyall; Michael R. Holbrook. 2018. "MERS-CoV pathogenesis and antiviral efficacy of licensed drugs in human monocyte-derived antigen-presenting cells." PLOS ONE 13, no. 3: e0194868.
Sexual transmission of Ebola virus (EBOV) has been demonstrated more than a year after recovery from the acute phase of Ebola virus disease (EVD). The mechanisms underlying EBOV persistence and sexual transmission are not currently understood. Using the acute macaque model of EVD, we hypothesized EBOV would infect the reproductive tissues and sought to localize the infection in these tissues using immunohistochemistry and transmission electron microscopy. In four female and eight male macaques that succumbed to EVD between 6 and 9 days after EBOV challenge, we demonstrate widespread EBOV infection of the interstitial tissues and endothelium in the ovary, uterus, testis, seminal vesicle, epididymis, and prostate gland, with minimal associated tissue immune response or organ pathology. Given the widespread involvement of EBOV in the reproductive tracts of both male and female macaques, it is reasonable to surmise that our understanding of the mechanisms underlying sexual transmission of EVD and persistence of EBOV in immune-privileged sites would be facilitated by the development of a nonhuman primate model in which the macaques survived past the acute stage into convalescence.
Donna L. Perry; Louis M. Huzella; John G. Bernbaum; Michael R. Holbrook; Peter B. Jahrling; Katie R. Hagen; Matthias J. Schnell; Reed F. Johnson. Ebola Virus Localization in the Macaque Reproductive Tract during Acute Ebola Virus Disease. The American Journal of Pathology 2018, 188, 550 -558.
AMA StyleDonna L. Perry, Louis M. Huzella, John G. Bernbaum, Michael R. Holbrook, Peter B. Jahrling, Katie R. Hagen, Matthias J. Schnell, Reed F. Johnson. Ebola Virus Localization in the Macaque Reproductive Tract during Acute Ebola Virus Disease. The American Journal of Pathology. 2018; 188 (3):550-558.
Chicago/Turabian StyleDonna L. Perry; Louis M. Huzella; John G. Bernbaum; Michael R. Holbrook; Peter B. Jahrling; Katie R. Hagen; Matthias J. Schnell; Reed F. Johnson. 2018. "Ebola Virus Localization in the Macaque Reproductive Tract during Acute Ebola Virus Disease." The American Journal of Pathology 188, no. 3: 550-558.