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Dr. Richard Bennett
NIAID/NIH

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0 Infectious Diseases
0 vaccine
0 Zoonotic diseases
0 animal models infection
0 Virology, Quality Control, BioManufacturing, vaccine, Viral Translational control,

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Journal article
Published: 12 May 2021 in Viruses
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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.

ACS Style

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 Style

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 (5):893.

Chicago/Turabian Style

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. 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.

Resource
Published: 06 November 2020 in Cell
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Summary Ebola virus (EBOV) causes epidemics with high mortality yet remains understudied due to the challenge of experimentation in high-containment and outbreak settings. Here, we used single-cell transcriptomics and CyTOF-based single-cell protein quantification to characterize peripheral immune cells during EBOV infection in rhesus monkeys. We obtained 100,000 transcriptomes and 15,000,000 protein profiles, finding that immature, proliferative monocyte-lineage cells with reduced antigen-presentation capacity replace conventional monocyte subsets, while lymphocytes upregulate apoptosis genes and decline in abundance. By quantifying intracellular viral RNA, we identify molecular determinants of tropism among circulating immune cells and examine temporal dynamics in viral and host gene expression. Within infected cells, EBOV downregulates STAT1 mRNA and interferon signaling, and it upregulates putative pro-viral genes (e.g., DYNLL1 and HSPA5), nominating pathways the virus manipulates for its replication. This study sheds light on EBOV tropism, replication dynamics, and elicited immune response and provides a framework for characterizing host-virus interactions under maximum containment.

ACS Style

Dylan Kotliar; Aaron E. Lin; James Logue; Travis K. Hughes; Nadine M. Khoury; Siddharth S. Raju; Marc H. Wadsworth; Han Chen; Jonathan R. Kurtz; Bonnie Dighero-Kemp; Zach B. Bjornson; Nilanjan Mukherjee; Brian A. Sellers; Nancy Tran; Matthew R. Bauer; Gordon C. Adams; Ricky Adams; John L. Rinn; Marta Melé; Stephen F. Schaffner; Garry P. Nolan; Kayla G. Barnes; Lisa E. Hensley; David R. McIlwain; Alex K. Shalek; Pardis C. Sabeti; Richard S. Bennett. Single-Cell Profiling of Ebola Virus Disease In Vivo Reveals Viral and Host Dynamics. Cell 2020, 183, 1383 -1401.e19.

AMA Style

Dylan Kotliar, Aaron E. Lin, James Logue, Travis K. Hughes, Nadine M. Khoury, Siddharth S. Raju, Marc H. Wadsworth, Han Chen, Jonathan R. Kurtz, Bonnie Dighero-Kemp, Zach B. Bjornson, Nilanjan Mukherjee, Brian A. Sellers, Nancy Tran, Matthew R. Bauer, Gordon C. Adams, Ricky Adams, John L. Rinn, Marta Melé, Stephen F. Schaffner, Garry P. Nolan, Kayla G. Barnes, Lisa E. Hensley, David R. McIlwain, Alex K. Shalek, Pardis C. Sabeti, Richard S. Bennett. Single-Cell Profiling of Ebola Virus Disease In Vivo Reveals Viral and Host Dynamics. Cell. 2020; 183 (5):1383-1401.e19.

Chicago/Turabian Style

Dylan Kotliar; Aaron E. Lin; James Logue; Travis K. Hughes; Nadine M. Khoury; Siddharth S. Raju; Marc H. Wadsworth; Han Chen; Jonathan R. Kurtz; Bonnie Dighero-Kemp; Zach B. Bjornson; Nilanjan Mukherjee; Brian A. Sellers; Nancy Tran; Matthew R. Bauer; Gordon C. Adams; Ricky Adams; John L. Rinn; Marta Melé; Stephen F. Schaffner; Garry P. Nolan; Kayla G. Barnes; Lisa E. Hensley; David R. McIlwain; Alex K. Shalek; Pardis C. Sabeti; Richard S. Bennett. 2020. "Single-Cell Profiling of Ebola Virus Disease In Vivo Reveals Viral and Host Dynamics." Cell 183, no. 5: 1383-1401.e19.

Journal article
Published: 14 July 2020 in Viruses
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Ongoing Ebola virus disease outbreaks in the Democratic Republic of the Congo follow the largest recorded outbreak in Western Africa (2013–2016). To combat outbreaks, testing of medical countermeasures (therapeutics or vaccines) requires a well-defined, reproducible, animal model. Here we present Ebola virus disease kinetics in 24 Chinese-origin rhesus monkeys exposed intramuscularly to a highly characterized, commercially available Kikwit Ebola virus Filovirus Animal Non-Clinical Group (FANG) stock. Until reaching predetermined clinical disease endpoint criteria, six animals underwent anesthesia for repeated clinical sampling and were compared to six that did not. Groups of three animals were euthanized and necropsied on days 3, 4, 5, and 6 post-exposure, respectively. In addition, three uninfected animals served as controls. Here, we present detailed characterization of clinical and laboratory disease kinetics and complete blood counts, serum chemistries, Ebola virus titers, and disease kinetics for future medical countermeasure (MCM) study design and control data. We measured no statistical difference in hematology, chemistry values, or time to clinical endpoint in animals that were anesthetized for clinical sampling during the acute disease compared to those that were not.

ACS Style

Richard S. Bennett; James Logue; David X. Liu; Rebecca J. Reeder; Krisztina B. Janosko; Donna L. Perry; Timothy K. Cooper; Russell Byrum; Danny Ragland; Marisa St. Claire; Ricky Adams; Tracey L. Burdette; Tyler M. Brady; Kyra Hadley; M. Colin Waters; Rebecca Shim; William Dowling; Jing Qin; Ian Crozier; Peter B. Jahrling; Lisa E. Hensley. Kikwit Ebola Virus Disease Progression in the Rhesus Monkey Animal Model. Viruses 2020, 12, 753 .

AMA Style

Richard S. Bennett, James Logue, David X. Liu, Rebecca J. Reeder, Krisztina B. Janosko, Donna L. Perry, Timothy K. Cooper, Russell Byrum, Danny Ragland, Marisa St. Claire, Ricky Adams, Tracey L. Burdette, Tyler M. Brady, Kyra Hadley, M. Colin Waters, Rebecca Shim, William Dowling, Jing Qin, Ian Crozier, Peter B. Jahrling, Lisa E. Hensley. Kikwit Ebola Virus Disease Progression in the Rhesus Monkey Animal Model. Viruses. 2020; 12 (7):753.

Chicago/Turabian Style

Richard S. Bennett; James Logue; David X. Liu; Rebecca J. Reeder; Krisztina B. Janosko; Donna L. Perry; Timothy K. Cooper; Russell Byrum; Danny Ragland; Marisa St. Claire; Ricky Adams; Tracey L. Burdette; Tyler M. Brady; Kyra Hadley; M. Colin Waters; Rebecca Shim; William Dowling; Jing Qin; Ian Crozier; Peter B. Jahrling; Lisa E. Hensley. 2020. "Kikwit Ebola Virus Disease Progression in the Rhesus Monkey Animal Model." Viruses 12, no. 7: 753.

Journal article
Published: 01 July 2020 in The American Journal of Pathology
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Zaire ebolavirus (EBOV) causes Ebola virus disease (EVD), which carries a fatality rate between 25% and 90% in humans. Liver pathology is a hallmark of terminal EVD; however, little is known about temporal disease progression. The authors utilized multiplexed fluorescent immunohistochemistry and in situ hybridization in combination with whole slide imaging and image analysis (IA) to quantitatively characterize temporospatial signatures of viral and host factors as related to EBOV pathogenesis. Eighteen rhesus monkeys euthanized between 3 and 8 days post-infection and 3 uninfected controls were enrolled in this study. Compared with semiquantitative histomorphologic ordinal scoring, quantitative IA was able to detect subtle and progressive features of early and terminal EVD that was not feasible with routine approaches. Sinusoidal macrophages were the earliest cells to respond to infection, expressing the proinflammatory cytokine gene IL6. IL6 expression was subsequently also observed in fibrovascular compartments. Interferon-stimulated gene-15 (ISG15), displayed an early, progressive, and ubiquitous signature with hybridization of both mesenchymal and epithelial compartments. ISG15 expression was prominent near infected cells, but not in infected cells, supporting the hypothesis that bystander cells produce a robust interferon gene response. This study contributes to our current understanding of early EVD progression and illustrates the value that digital pathology and quantitative IA serve in infectious disease research.

ACS Style

Alexandra Greenberg; Bertrand R. Huber; David X. Liu; James Logue; Amanda M.W. Hischak; Randy J. Hart; Maureen Abbott; Nejra Isic; Yohei Hisada; Nigel Mackman; Richard S. Bennett; Lisa E. Hensley; John H. Connor; Nicholas A. Crossland. Quantification of Viral and Host Biomarkers in the Liver of Rhesus Macaques. The American Journal of Pathology 2020, 190, 1449 -1460.

AMA Style

Alexandra Greenberg, Bertrand R. Huber, David X. Liu, James Logue, Amanda M.W. Hischak, Randy J. Hart, Maureen Abbott, Nejra Isic, Yohei Hisada, Nigel Mackman, Richard S. Bennett, Lisa E. Hensley, John H. Connor, Nicholas A. Crossland. Quantification of Viral and Host Biomarkers in the Liver of Rhesus Macaques. The American Journal of Pathology. 2020; 190 (7):1449-1460.

Chicago/Turabian Style

Alexandra Greenberg; Bertrand R. Huber; David X. Liu; James Logue; Amanda M.W. Hischak; Randy J. Hart; Maureen Abbott; Nejra Isic; Yohei Hisada; Nigel Mackman; Richard S. Bennett; Lisa E. Hensley; John H. Connor; Nicholas A. Crossland. 2020. "Quantification of Viral and Host Biomarkers in the Liver of Rhesus Macaques." The American Journal of Pathology 190, no. 7: 1449-1460.

Preprint content
Published: 13 June 2020
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SummaryEbola virus (EBOV) causes epidemics with high case fatality rates, yet remains understudied due to the challenge of experimentation in high-containment and outbreak settings. To better understand EBOV infection in vivo, we used single-cell transcriptomics and CyTOF-based single-cell protein quantification to characterize peripheral immune cell activity during EBOV infection in rhesus monkeys. We obtained 100,000 transcriptomes and 15,000,000 protein profiles, providing insight into pathogenesis. We find that immature, proliferative monocyte-lineage cells with reduced antigen presentation capacity replace conventional circulating monocyte subsets within days of infection, while lymphocytes upregulate apoptosis genes and decline in abundance. By quantifying viral RNA abundance in individual cells, we identify molecular determinants of tropism and examine temporal dynamics in viral and host gene expression. Within infected cells, we observe that EBOV down-regulates STAT1 mRNA and interferon signaling, and up-regulates putative pro-viral genes (e.g., DYNLL1 and HSPA5), nominating cellular pathways the virus manipulates for its replication. Overall, this study sheds light on EBOV tropism, replication dynamics, and elicited immune response, and provides a framework for characterizing interactions between hosts and emerging viruses in a maximum containment setting.

ACS Style

Dylan Kotliar; Aaron E. Lin; James Logue; Travis K. Hughes; Nadine M. Khoury; Siddharth S. Raju; Marc H. Wadsworth; Han Chen; Jonathan R. Kurtz; Bonnie Dighero-Kemp; Zach B. Bjornson; Nilanjan Mukherjee; Brian A. Sellers; Nancy Tran; Matthew R. Bauer; Gordon C. Adams; Ricky Adams; John L. Rinn; Marta Melé; Garry P. Nolan; Kayla G. Barnes; Lisa E. Hensley; David R. McIlwain; Alex K. Shalek; Pardis C. Sabeti; Richard S. Bennett. Single-cell profiling of Ebola virus infection in vivo reveals viral and host transcriptional dynamics. 2020, 1 .

AMA Style

Dylan Kotliar, Aaron E. Lin, James Logue, Travis K. Hughes, Nadine M. Khoury, Siddharth S. Raju, Marc H. Wadsworth, Han Chen, Jonathan R. Kurtz, Bonnie Dighero-Kemp, Zach B. Bjornson, Nilanjan Mukherjee, Brian A. Sellers, Nancy Tran, Matthew R. Bauer, Gordon C. Adams, Ricky Adams, John L. Rinn, Marta Melé, Garry P. Nolan, Kayla G. Barnes, Lisa E. Hensley, David R. McIlwain, Alex K. Shalek, Pardis C. Sabeti, Richard S. Bennett. Single-cell profiling of Ebola virus infection in vivo reveals viral and host transcriptional dynamics. . 2020; ():1.

Chicago/Turabian Style

Dylan Kotliar; Aaron E. Lin; James Logue; Travis K. Hughes; Nadine M. Khoury; Siddharth S. Raju; Marc H. Wadsworth; Han Chen; Jonathan R. Kurtz; Bonnie Dighero-Kemp; Zach B. Bjornson; Nilanjan Mukherjee; Brian A. Sellers; Nancy Tran; Matthew R. Bauer; Gordon C. Adams; Ricky Adams; John L. Rinn; Marta Melé; Garry P. Nolan; Kayla G. Barnes; Lisa E. Hensley; David R. McIlwain; Alex K. Shalek; Pardis C. Sabeti; Richard S. Bennett. 2020. "Single-cell profiling of Ebola virus infection in vivo reveals viral and host transcriptional dynamics." , no. : 1.

Journal article
Published: 28 May 2020 in The American Journal of Pathology
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The most commonly reported symptom of post-Ebola virus disease syndrome in survivors is arthralgia, yet involvement of the joints in acute or convalescent Ebola virus infection has not been well-characterized in human patients or animal models. By immunohistochemistry, we found that the lining synovial intima of the stifle (knee) is a target for acute infection by Ebola virus/Kikwit, Ebola virus/Makona-C05, and Marburg virus/Angola in the rhesus macaque model. Further, we demonstrate by histology, immunohistochemistry, RNAscope in situ hybridization, and transmission electron microscopy that synoviocytes of the stifle, shoulder, and hip are a target for mouse-adapted Ebola virus/Yambuku-Mayinga infection during acute disease in rhesus macaques. A time course of infection study with Ebola virus/Kikwit found large joint synovium became immunopositive beginning on post-infection day 6. In total, the synovium of 28/30 rhesus macaques with terminal filovirus disease had evidence of infection (64/96 joints examined). By immunofluorescence, infected cell types included both CD68+ type A (macrophage-like) synoviocytes and CD44+ type B (fibroblast-like) synoviocytes. Cultured primary human fibroblast-like synoviocytes were permissive to infection with Ebola and Marburg viruses in vitro. Because synovial joints include immune privileged sites, these findings are significant for future investigations of filovirus pathogenesis and persistence as well as arthralgias in acute and convalescent filovirus disease.

ACS Style

Timothy K. Cooper; James Logue; David X. Liu; Donna L. Perry; Randy J. Hart; Amanda M.W. Hischak; John G. Bernbaum; Dawn M. Gerhardt; Oscar Rojas; J. Kyle Bohannon; Katie R. Hagen; Reed F. Johnson; Ian Crozier; Peter B. Jahrling; Lisa Hensley; Richard S. Bennett. Filoviruses Infect Rhesus Macaque Synoviocytes in Vivo and Primary Human Synoviocytes in Vitro. The American Journal of Pathology 2020, 190, 1867 -1880.

AMA Style

Timothy K. Cooper, James Logue, David X. Liu, Donna L. Perry, Randy J. Hart, Amanda M.W. Hischak, John G. Bernbaum, Dawn M. Gerhardt, Oscar Rojas, J. Kyle Bohannon, Katie R. Hagen, Reed F. Johnson, Ian Crozier, Peter B. Jahrling, Lisa Hensley, Richard S. Bennett. Filoviruses Infect Rhesus Macaque Synoviocytes in Vivo and Primary Human Synoviocytes in Vitro. The American Journal of Pathology. 2020; 190 (9):1867-1880.

Chicago/Turabian Style

Timothy K. Cooper; James Logue; David X. Liu; Donna L. Perry; Randy J. Hart; Amanda M.W. Hischak; John G. Bernbaum; Dawn M. Gerhardt; Oscar Rojas; J. Kyle Bohannon; Katie R. Hagen; Reed F. Johnson; Ian Crozier; Peter B. Jahrling; Lisa Hensley; Richard S. Bennett. 2020. "Filoviruses Infect Rhesus Macaque Synoviocytes in Vivo and Primary Human Synoviocytes in Vitro." The American Journal of Pathology 190, no. 9: 1867-1880.

Journal article
Published: 01 October 2019 in Radiology Case Reports
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During an infectious disease modeling study, a rhesus macaque (Macaca mulatta), experienced acute transient tachypnea including transient severe motion during the 70-second phases of serial contrast-enhanced magnetic resonance imaging of the abdomen. This same animal experienced transient severe motion during all but 2 of the 8 scans of the year-long study. This animal was the only animal in the study (1 of 12) to have this reaction to gadoxetate; the animal also vomited after the contrast injection once on day 146 of the study. On day 86, a different contrast agent (gadobutrol) was used, and the reaction did not occur. No treatment was required for any conditions relating to the reaction due to the self-limited nature. This type of reaction has not yet been reported in veterinary subjects before and is likely to be idiosyncratic after first exposure. However, this reaction should not be life threatening, and other contrast agents can be used if acute transient tachypnea does occur.

ACS Style

Nina M. Aiosa; Joseph Laux; Oscar Rojas; Richard Bennett; James Logue; Ji Hyun Lee; Laura Bollinger; Russell Byrum; Marisa St. Claire; Irwin M. Feuerstein. Acute transient tachypnea following gadoxetate administration in a rhesus macaque during contrast-enhanced magnetic resonance imaging. Radiology Case Reports 2019, 14, 1272 -1275.

AMA Style

Nina M. Aiosa, Joseph Laux, Oscar Rojas, Richard Bennett, James Logue, Ji Hyun Lee, Laura Bollinger, Russell Byrum, Marisa St. Claire, Irwin M. Feuerstein. Acute transient tachypnea following gadoxetate administration in a rhesus macaque during contrast-enhanced magnetic resonance imaging. Radiology Case Reports. 2019; 14 (10):1272-1275.

Chicago/Turabian Style

Nina M. Aiosa; Joseph Laux; Oscar Rojas; Richard Bennett; James Logue; Ji Hyun Lee; Laura Bollinger; Russell Byrum; Marisa St. Claire; Irwin M. Feuerstein. 2019. "Acute transient tachypnea following gadoxetate administration in a rhesus macaque during contrast-enhanced magnetic resonance imaging." Radiology Case Reports 14, no. 10: 1272-1275.

Research article
Published: 27 August 2019 in PLOS ONE
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Antibody titers against a viral pathogen are typically measured using an antigen binding assay, such as an enzyme-linked immunosorbent assay (ELISA), which only measures the ability of antibodies to identify a viral antigen of interest. Neutralization assays measure the presence of virus-neutralizing antibodies in a sample. Traditional neutralization assays, such as the plaque reduction neutralization test (PRNT), are often difficult to use on a large scale due to being both labor and resource intensive. Here we describe an Ebola virus fluorescence reduction neutralization assay (FRNA), which tests for neutralizing antibodies, that requires only a small volume of sample in a 96-well format and is easy to automate. The readout of the FRNA is the percentage of Ebola virus-infected cells measured with an optical reader or overall chemiluminescence that can be generated by multiple reading platforms. Using blinded human clinical samples (EVD survivors or contacts) obtained in Liberia during the 2013-2016 Ebola virus disease outbreak, we demonstrate there was a high degree of agreement between the FRNA-measured antibody titers and the Filovirus Animal Non-clinical Group (FANG) ELISA titers with the FRNA providing information on the neutralizing capabilities of the antibodies.

ACS Style

Elena N. Postnikova; James Pettitt; Collin J. Van Ryn; Michael R. Holbrook; Laura Bollinger; Shuǐqìng Yú; Yingyun Cai; Janie Liang; Michael C. Sneller; Peter B. Jahrling; Lisa Hensley; Jens H. Kuhn; Mosoka P. Fallah; Richard S. Bennett; Cavan Reilly. Scalable, semi-automated fluorescence reduction neutralization assay for qualitative assessment of Ebola virus-neutralizing antibodies in human clinical samples. PLOS ONE 2019, 14, e0221407 .

AMA Style

Elena N. Postnikova, James Pettitt, Collin J. Van Ryn, Michael R. Holbrook, Laura Bollinger, Shuǐqìng Yú, Yingyun Cai, Janie Liang, Michael C. Sneller, Peter B. Jahrling, Lisa Hensley, Jens H. Kuhn, Mosoka P. Fallah, Richard S. Bennett, Cavan Reilly. Scalable, semi-automated fluorescence reduction neutralization assay for qualitative assessment of Ebola virus-neutralizing antibodies in human clinical samples. PLOS ONE. 2019; 14 (8):e0221407.

Chicago/Turabian Style

Elena N. Postnikova; James Pettitt; Collin J. Van Ryn; Michael R. Holbrook; Laura Bollinger; Shuǐqìng Yú; Yingyun Cai; Janie Liang; Michael C. Sneller; Peter B. Jahrling; Lisa Hensley; Jens H. Kuhn; Mosoka P. Fallah; Richard S. Bennett; Cavan Reilly. 2019. "Scalable, semi-automated fluorescence reduction neutralization assay for qualitative assessment of Ebola virus-neutralizing antibodies in human clinical samples." PLOS ONE 14, no. 8: e0221407.

Comparative study
Published: 16 February 2019 in Viruses
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Following the largest Ebola virus disease outbreak from 2013 to 2016, viral RNA has been detected in survivors from semen and breast milk long after disease recovery. However, as there have been few cases of sexual transmission, it is unclear whether every RNA positive fluid sample contains infectious virus. Virus isolation, typically using cell culture or animal models, can serve as a tool to determine the infectivity of patient samples. However, the sensitivity of these methods has not been assessed for the Ebola virus isolate, Makona. Described here is an efficiency comparison of Ebola virus Makona isolation using Vero E6, Huh-7, monocyte-derived macrophage cells, and suckling laboratory mice. Isolation sensitivity was similar in all methods tested. Laboratory mice and Huh-7 cells were less affected by toxicity from breast milk than Vero E6 and MDM cells. However, the advantages associated with isolation in Huh-7 cells over laboratory mice, including cost effectiveness, sample volume preservation, and a reduction in animal use, make Huh-7 cells the preferred substrate tested for Ebola virus Makona isolation.

ACS Style

James Logue; Walter Vargas Licona; Timothy K. Cooper; Becky Reeder; Russel Byrum; Jing Qin; Nicole Deiuliis Murphy; Yu Cong; Amanda Bonilla; Jennifer Sword; Wade Weaver; Gregory Kocher; Gene G. Olinger; Peter B. Jahrling; Lisa E. Hensley; Richard S. Bennett. Ebola Virus Isolation Using Huh-7 Cells has Methodological Advantages and Similar Sensitivity to Isolation Using Other Cell Types and Suckling BALB/c Laboratory Mice. Viruses 2019, 11, 161 .

AMA Style

James Logue, Walter Vargas Licona, Timothy K. Cooper, Becky Reeder, Russel Byrum, Jing Qin, Nicole Deiuliis Murphy, Yu Cong, Amanda Bonilla, Jennifer Sword, Wade Weaver, Gregory Kocher, Gene G. Olinger, Peter B. Jahrling, Lisa E. Hensley, Richard S. Bennett. Ebola Virus Isolation Using Huh-7 Cells has Methodological Advantages and Similar Sensitivity to Isolation Using Other Cell Types and Suckling BALB/c Laboratory Mice. Viruses. 2019; 11 (2):161.

Chicago/Turabian Style

James Logue; Walter Vargas Licona; Timothy K. Cooper; Becky Reeder; Russel Byrum; Jing Qin; Nicole Deiuliis Murphy; Yu Cong; Amanda Bonilla; Jennifer Sword; Wade Weaver; Gregory Kocher; Gene G. Olinger; Peter B. Jahrling; Lisa E. Hensley; Richard S. Bennett. 2019. "Ebola Virus Isolation Using Huh-7 Cells has Methodological Advantages and Similar Sensitivity to Isolation Using Other Cell Types and Suckling BALB/c Laboratory Mice." Viruses 11, no. 2: 161.

Journal article
Published: 31 August 2018 in Virology Journal
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Currently, no FDA-approved vaccines or treatments are available for Ebola virus disease (EVD), and therapy remains largely supportive. Ebola virus (EBOV) has broad tissue tropism and can infect a variety of cells including epithelial cells. Epithelial cells differ from most other cell types by their polarized phenotype and barrier function. In polarized cells, the apical and basolateral membrane domains are demarcated by tight junctions, and specialized sorting machinery, which results in a difference in composition between the two membrane domains. These specialized sorting functions can have important consequences for viral infections. Differential localization of a viral receptor can restrict virus entry to a particular membrane while polarized sorting can lead to a vectorial virus release. The present study investigated the impact of cell polarity on EBOV infection. Characteristics of EBOV infection in polarized cells were evaluated in the polarized Caco-2 model grown on semipermeable transwells. Transepithelial resistance (TEER), which is a function of tight junctions, was used to assess epithelial cell polarization. EBOV infection was assessed with immunofluorescence microscopy and qPCR. Statistical significance was calculated using one-way ANOVA and significance was set at p < 0.05. Our data indicate that EBOV preferentially infects cells from the basolateral route, and this preference may be influenced by the resistance across the Caco-2 monolayer. Infection occurs without changes in cellular permeability. Further, our data show that basolateral infection bias may be dependent on polarized distribution of heparan sulfate, a known viral attachment factor. Treatment with iota-carrageenan, or heparin lyase, which interrupts viral interaction with cellular heparan sulfate, significantly reduced cell susceptibility to basolateral infection, likely by inhibiting virus attachment. Our results show cell polarity has an impact on EBOV infection. EBOV preferentially infects polarized cells through the basolateral route. Access to heparan sulfate is an important factor during basolateral infection and blocking interaction of cellular heparan sulfate with virus leads to significant inhibition of basolateral infection in the polarized Caco-2 cell model.

ACS Style

Manasi Tamhankar; Dawn M. Gerhardt; Richard S. Bennett; Nicole Murphy; Peter B. Jahrling; Jean L. Patterson. Heparan sulfate is an important mediator of Ebola virus infection in polarized epithelial cells. Virology Journal 2018, 15, 1 -12.

AMA Style

Manasi Tamhankar, Dawn M. Gerhardt, Richard S. Bennett, Nicole Murphy, Peter B. Jahrling, Jean L. Patterson. Heparan sulfate is an important mediator of Ebola virus infection in polarized epithelial cells. Virology Journal. 2018; 15 (1):1-12.

Chicago/Turabian Style

Manasi Tamhankar; Dawn M. Gerhardt; Richard S. Bennett; Nicole Murphy; Peter B. Jahrling; Jean L. Patterson. 2018. "Heparan sulfate is an important mediator of Ebola virus infection in polarized epithelial cells." Virology Journal 15, no. 1: 1-12.

Comparative study
Published: 23 August 2018 in The Journal of Infectious Diseases
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The domestic ferret is a uniformly lethal model of infection for 3 species of Ebolavirus known to be pathogenic in humans. Reagents to systematically analyze the ferret host response to infection are lacking; however, the recent publication of a draft ferret genome has opened the potential for transcriptional analysis of ferret models of disease. In this work, we present comparative analysis of longitudinally sampled blood taken from ferrets and nonhuman primates infected with lethal doses of the Makona variant of Zaire ebolavirus. Strong induction of proinflammatory and prothrombotic signaling programs were present in both ferrets and nonhuman primates, and both transcriptomes were similar to previously published datasets of fatal cases of human Ebola virus infection.

ACS Style

Robert W Cross; Emily Speranza; Viktoriya Borisevich; Steven G Widen; Thomas G Wood; Rebecca S Shim; Ricky D Adams; Dawn M Gerhardt; Richard S Bennett; Anna Honko; Joshua Johnson; Lisa Hensley; Thomas W Geisbert; John H Connor. Comparative Transcriptomics in Ebola Makona-Infected Ferrets, Nonhuman Primates, and Humans. The Journal of Infectious Diseases 2018, 218, S486 -S495.

AMA Style

Robert W Cross, Emily Speranza, Viktoriya Borisevich, Steven G Widen, Thomas G Wood, Rebecca S Shim, Ricky D Adams, Dawn M Gerhardt, Richard S Bennett, Anna Honko, Joshua Johnson, Lisa Hensley, Thomas W Geisbert, John H Connor. Comparative Transcriptomics in Ebola Makona-Infected Ferrets, Nonhuman Primates, and Humans. The Journal of Infectious Diseases. 2018; 218 ():S486-S495.

Chicago/Turabian Style

Robert W Cross; Emily Speranza; Viktoriya Borisevich; Steven G Widen; Thomas G Wood; Rebecca S Shim; Ricky D Adams; Dawn M Gerhardt; Richard S Bennett; Anna Honko; Joshua Johnson; Lisa Hensley; Thomas W Geisbert; John H Connor. 2018. "Comparative Transcriptomics in Ebola Makona-Infected Ferrets, Nonhuman Primates, and Humans." The Journal of Infectious Diseases 218, no. : S486-S495.

Journal article
Published: 13 August 2018 in The Journal of Infectious Diseases
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One year after a Zaire ebolavirus (EBOV) outbreak occurred in the Boende Health Zone of the Democratic Republic of the Congo during 2014, we sought to determine the breadth of immune response against diverse filoviruses including EBOV, Bundibugyo (BDBV), Sudan (SUDV), and Marburg (MARV) viruses. After assessing the 15 survivors, 5 individuals demonstrated some degree of reactivity to multiple ebolavirus species and, in some instances, Marburg virus. All 5 of these survivors had immunoreactivity to EBOV glycoprotein (GP) and EBOV VP40, and 4 had reactivity to EBOV nucleoprotein (NP). Three of these survivors showed serologic responses to the 3 species of ebolavirus GPs tested (EBOV, BDBV, SUDV). All 5 samples also exhibited ability to neutralize EBOV using live virus, in a plaque reduction neutralization test. Remarkably, 3 of these EBOV survivors had plasma antibody responses to MARV GP. In pseudovirus neutralization assays, serum antibodies from a subset of these survivors also neutralized EBOV, BDBV, SUDV, and Taï Forest virus as well as MARV. Collectively, these findings suggest that some survivors of naturally acquired ebolavirus infection mount not only a pan-ebolavirus response, but also in less frequent cases, a pan-filovirus neutralizing response.

ACS Style

Matthew S Bramble; Nicole Hoff; Pavlo Gilchuk; Patrick Mukadi; Kai Lu; Reena H Doshi; Imke Steffen; Bradly P Nicholson; Allen Lipson; Neerja Vashist; Cyrus Sinai; D’Andre Spencer; Garrard Olinger; Emile Okitolonda Wemakoy; Benoit Kebela Illunga; James Pettitt; James Logue; Jonathan Marchand; Justin Varughese; Richard S Bennett; Peter Jahrling; Guy Cavet; Tito Serafini; Erica Ollmann Saphire; Eric Vilain; Jean Jacques Muyembe-Tamfum; Lisa E Hensely; Graham Simmons; James Crowe; Anne W Rimoin. Pan-Filovirus Serum Neutralizing Antibodies in a Subset of Congolese Ebolavirus Infection Survivors. The Journal of Infectious Diseases 2018, 218, 1929 -1936.

AMA Style

Matthew S Bramble, Nicole Hoff, Pavlo Gilchuk, Patrick Mukadi, Kai Lu, Reena H Doshi, Imke Steffen, Bradly P Nicholson, Allen Lipson, Neerja Vashist, Cyrus Sinai, D’Andre Spencer, Garrard Olinger, Emile Okitolonda Wemakoy, Benoit Kebela Illunga, James Pettitt, James Logue, Jonathan Marchand, Justin Varughese, Richard S Bennett, Peter Jahrling, Guy Cavet, Tito Serafini, Erica Ollmann Saphire, Eric Vilain, Jean Jacques Muyembe-Tamfum, Lisa E Hensely, Graham Simmons, James Crowe, Anne W Rimoin. Pan-Filovirus Serum Neutralizing Antibodies in a Subset of Congolese Ebolavirus Infection Survivors. The Journal of Infectious Diseases. 2018; 218 (12):1929-1936.

Chicago/Turabian Style

Matthew S Bramble; Nicole Hoff; Pavlo Gilchuk; Patrick Mukadi; Kai Lu; Reena H Doshi; Imke Steffen; Bradly P Nicholson; Allen Lipson; Neerja Vashist; Cyrus Sinai; D’Andre Spencer; Garrard Olinger; Emile Okitolonda Wemakoy; Benoit Kebela Illunga; James Pettitt; James Logue; Jonathan Marchand; Justin Varughese; Richard S Bennett; Peter Jahrling; Guy Cavet; Tito Serafini; Erica Ollmann Saphire; Eric Vilain; Jean Jacques Muyembe-Tamfum; Lisa E Hensely; Graham Simmons; James Crowe; Anne W Rimoin. 2018. "Pan-Filovirus Serum Neutralizing Antibodies in a Subset of Congolese Ebolavirus Infection Survivors." The Journal of Infectious Diseases 218, no. 12: 1929-1936.

Journal article
Published: 16 July 2018 in The Journal of Infectious Diseases
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Transchromosomic bovines (Tc-bovines) adaptively produce fully human polyclonal immunoglobulin (Ig)G antibodies after exposure to immunogenic antigen(s). The National Interagency Confederation for Biological Research and collaborators rapidly produced and then evaluated anti-Ebola virus IgG immunoglobulins (collectively termed SAB-139) purified from Tc-bovine plasma after sequential hyperimmunization with an Ebola virus Makona isolate glycoprotein nanoparticle vaccine. SAB-139 was characterized by several in vitro production, research, and clinical level assays using wild-type Makona-C05 or recombinant virus/antigens from different Ebola virus variants. SAB-139 potently activates natural killer cells, monocytes, and peripheral blood mononuclear cells and has high-binding avidity demonstrated by surface plasmon resonance. SAB-139 has similar concentrations of galactose-α-1,3-galactose carbohydrates compared with human-derived intravenous Ig, and the IgG1 subclass antibody is predominant. All rhesus macaques infected with Ebola virus/H.sapiens-tc/GIN/2014/Makona-C05 and treated with sufficient SAB-139 at 1 day (n = 6) or 3 days (n = 6) postinfection survived versus 0% of controls. This study demonstrates that Tc-bovines can produce pathogen-specific human Ig to prevent and/or treat patients when an emerging infectious disease either threatens to or becomes an epidemic.

ACS Style

Thomas Luke; Richard Bennett; Dawn M Gerhardt; Tracey Burdette; Elena Postnikova; Steven Mazur; Anna Honko; Nicholas Oberlander; Russell Byrum; Dan Ragland; Marisa St. Claire; Krisztina B Janosko; Gale Smith; Gregory Glenn; Jay Hooper; John Dye; Subhamoy Pal; Kimberly A Bishop-Lilly; Theron Hamilton; Kenneth Frey; Laura Bollinger; Jiro Wada; Hua Wu; Jin-An Jiao; Gene Olinger; Bronwyn Gunn; Galit Alter; Surender Khurana; Lisa Hensley; Eddie Sullivan; Peter B Jahrling. Fully Human Immunoglobulin G From Transchromosomic Bovines Treats Nonhuman Primates Infected With Ebola Virus Makona Isolate. The Journal of Infectious Diseases 2018, 218, S636 -S648.

AMA Style

Thomas Luke, Richard Bennett, Dawn M Gerhardt, Tracey Burdette, Elena Postnikova, Steven Mazur, Anna Honko, Nicholas Oberlander, Russell Byrum, Dan Ragland, Marisa St. Claire, Krisztina B Janosko, Gale Smith, Gregory Glenn, Jay Hooper, John Dye, Subhamoy Pal, Kimberly A Bishop-Lilly, Theron Hamilton, Kenneth Frey, Laura Bollinger, Jiro Wada, Hua Wu, Jin-An Jiao, Gene Olinger, Bronwyn Gunn, Galit Alter, Surender Khurana, Lisa Hensley, Eddie Sullivan, Peter B Jahrling. Fully Human Immunoglobulin G From Transchromosomic Bovines Treats Nonhuman Primates Infected With Ebola Virus Makona Isolate. The Journal of Infectious Diseases. 2018; 218 (suppl_5):S636-S648.

Chicago/Turabian Style

Thomas Luke; Richard Bennett; Dawn M Gerhardt; Tracey Burdette; Elena Postnikova; Steven Mazur; Anna Honko; Nicholas Oberlander; Russell Byrum; Dan Ragland; Marisa St. Claire; Krisztina B Janosko; Gale Smith; Gregory Glenn; Jay Hooper; John Dye; Subhamoy Pal; Kimberly A Bishop-Lilly; Theron Hamilton; Kenneth Frey; Laura Bollinger; Jiro Wada; Hua Wu; Jin-An Jiao; Gene Olinger; Bronwyn Gunn; Galit Alter; Surender Khurana; Lisa Hensley; Eddie Sullivan; Peter B Jahrling. 2018. "Fully Human Immunoglobulin G From Transchromosomic Bovines Treats Nonhuman Primates Infected With Ebola Virus Makona Isolate." The Journal of Infectious Diseases 218, no. suppl_5: S636-S648.

Journal article
Published: 16 July 2018 in Journal of Infectious Diseases
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At the onset of the 2013–2016 epidemic of Ebola virus disease (EVD), no vaccine or antiviral medication was approved for treatment. Therefore, considerable efforts were directed towards the concept of drug repurposing or repositioning. Amiodarone, an approved multi-ion channel blocker for the treatment of cardiac arrhythmia, was reported to inhibit filovirus entry in vitro. Compassionate use of amiodarone in EVD patients indicated a possible survival benefit. In support of further clinical testing, we confirmed anti-Ebola virus activity of amiodarone in different cell types. Despite promising in vitro results, amiodarone failed to protect guinea pigs from a lethal dose of Ebola virus.

ACS Style

Julie Dyall; Joshua C Johnson; Brit J Hart; Elena Postnikova; Yu Cong; Huanying Zhou; Dawn M Gerhardt; Julia Michelotti; Anna Honko; Steven Kern; Lisa Evans Dewald; Kathleen G O’Loughlin; Carol E Green; Jon C Mirsalis; Richard Bennett; Gene Olinger; Peter B Jahrling; Lisa Hensley. In Vitro and In Vivo Activity of Amiodarone Against Ebola Virus. Journal of Infectious Diseases 2018, 218, S592 -S596.

AMA Style

Julie Dyall, Joshua C Johnson, Brit J Hart, Elena Postnikova, Yu Cong, Huanying Zhou, Dawn M Gerhardt, Julia Michelotti, Anna Honko, Steven Kern, Lisa Evans Dewald, Kathleen G O’Loughlin, Carol E Green, Jon C Mirsalis, Richard Bennett, Gene Olinger, Peter B Jahrling, Lisa Hensley. In Vitro and In Vivo Activity of Amiodarone Against Ebola Virus. Journal of Infectious Diseases. 2018; 218 (suppl_5):S592-S596.

Chicago/Turabian Style

Julie Dyall; Joshua C Johnson; Brit J Hart; Elena Postnikova; Yu Cong; Huanying Zhou; Dawn M Gerhardt; Julia Michelotti; Anna Honko; Steven Kern; Lisa Evans Dewald; Kathleen G O’Loughlin; Carol E Green; Jon C Mirsalis; Richard Bennett; Gene Olinger; Peter B Jahrling; Lisa Hensley. 2018. "In Vitro and In Vivo Activity of Amiodarone Against Ebola Virus." Journal of Infectious Diseases 218, no. suppl_5: S592-S596.

Journal article
Published: 25 June 2018 in Journal of Infectious Diseases
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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.

ACS Style

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 Style

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 (suppl_5):S672-S678.

Chicago/Turabian Style

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. 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.

Research article
Published: 19 June 2018 in PLOS ONE
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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.

ACS Style

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 Style

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 (6):e0199534.

Chicago/Turabian Style

Kenneth 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.

Preprint
Published: 15 June 2018
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Defective viral genomes of the copy-back type (cbDVGs) are the primary initiators of the antiviral immune response during infection with respiratory syncytial virus (RSV) both in vitro and in vivo. However, the mechanism governing cbDVG generation remains unknown, thereby limiting our ability to manipulate cbDVG content in order to modulate the host response to infection. Here we report a specific genomic signal that mediates the generation of RSV cbDVGs. Using a customized bioinformatics tool, we identified regions in the RSV genome frequently used to generate cbDVGs during infection. We then created a minigenome system to validate the function of one of these sequences and to determine if specific nucleotides were essential for cbDVG generation at that position. Further, we created a recombinant virus that selectively produced a specific cbDVG based on variations introduced in this sequence. The identified sequence was also found as a common site for cbDVG generation during natural RSV infections, and common cbDVGs generated at this sequence were found among samples from various infected patients. These data demonstrate that sequences encoded in the viral genome are critical determinants of the location of cbDVG generation and, therefore, this is not a stochastic process. Most importantly, these findings open the possibility of genetically manipulating cbDVG formation to modulate infection outcome.Author summaryCopy-back defective viral genomes (cbDVGs) regulate infection and pathogenesis of Mononegavirales. cbDVG are believed to arise from random errors that occur during virus replication and the predominant hypothesis is that the viral polymerase is the main driver of cbDVG generation. Here we describe a specific genomic sequence in the RSV genome that is necessary for the generation of a large proportion of the cbDVG population present during infection. We identified specific nucleotides that when modified altered cbDVG generation at this position, and we created a recombinant virus that selectively produced cbDVGs based on mutations in this sequence. These data demonstrate that the generation of RSV cbDVGs is regulated by specific viral sequences and that these sequences can be manipulated to alter the content and quality of cbDVG generated during infection.

ACS Style

Yan Sun; Eun Ji Kim; Sébastien A. Felt; Louis J. Taylor; Divyansh Agarwal; Gregory R. Grant; Carolina B. López; Emily Speranza; Dawn Gerhardt; Richard Bennett; John Connor. A specific sequence in the genome of respiratory syncytial virus regulates the generation of copy-back defective viral genomes. 2018, 349001 .

AMA Style

Yan Sun, Eun Ji Kim, Sébastien A. Felt, Louis J. Taylor, Divyansh Agarwal, Gregory R. Grant, Carolina B. López, Emily Speranza, Dawn Gerhardt, Richard Bennett, John Connor. A specific sequence in the genome of respiratory syncytial virus regulates the generation of copy-back defective viral genomes. . 2018; ():349001.

Chicago/Turabian Style

Yan Sun; Eun Ji Kim; Sébastien A. Felt; Louis J. Taylor; Divyansh Agarwal; Gregory R. Grant; Carolina B. López; Emily Speranza; Dawn Gerhardt; Richard Bennett; John Connor. 2018. "A specific sequence in the genome of respiratory syncytial virus regulates the generation of copy-back defective viral genomes." , no. : 349001.

Research article
Published: 22 March 2018 in PLoS ONE
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Identifying effective antivirals for treating Ebola virus disease (EVD) and minimizing transmission of such disease is critical. A variety of cell-based assays have been developed for evaluating compounds for activity against Ebola virus. However, very few reports discuss the variable assay conditions that can affect the results obtained from these drug screens. Here, we describe variable conditions tested during the development of our cell-based drug screen assays designed to identify compounds with anti-Ebola virus activity using established cell lines and human primary cells. The effect of multiple assay readouts and variable assay conditions, including virus input, time of infection, and the cell passage number, were compared, and the impact on the effective concentration for 50% and/ or 90% inhibition (EC50, EC90) was evaluated using the FDA-approved compound, toremifene citrate. In these studies, we show that altering cell-based assay conditions can have an impact on apparent drug potency as measured by the EC50. These results further support the importance of developing standard operating procedures for generating reliable and reproducible in vitro data sets for potential antivirals.

ACS Style

Elena Postnikova; Yu Cong; Lisa Evans Dewald; Julie Dyall; Shuiqing Yu; Brit J. Hart; Huanying Zhou; Robin Gross; James Logue; Yingyun Cai; Nicole Deiuliis; Julia Michelotti; Anna Honko; Richard Bennett; Michael R. Holbrook; Gene Olinger; Lisa Hensley; Peter B. Jahrling. Testing therapeutics in cell-based assays: Factors that influence the apparent potency of drugs. PLoS ONE 2018, 13, e0194880 .

AMA Style

Elena Postnikova, Yu Cong, Lisa Evans Dewald, Julie Dyall, Shuiqing Yu, Brit J. Hart, Huanying Zhou, Robin Gross, James Logue, Yingyun Cai, Nicole Deiuliis, Julia Michelotti, Anna Honko, Richard Bennett, Michael R. Holbrook, Gene Olinger, Lisa Hensley, Peter B. Jahrling. Testing therapeutics in cell-based assays: Factors that influence the apparent potency of drugs. PLoS ONE. 2018; 13 (3):e0194880.

Chicago/Turabian Style

Elena Postnikova; Yu Cong; Lisa Evans Dewald; Julie Dyall; Shuiqing Yu; Brit J. Hart; Huanying Zhou; Robin Gross; James Logue; Yingyun Cai; Nicole Deiuliis; Julia Michelotti; Anna Honko; Richard Bennett; Michael R. Holbrook; Gene Olinger; Lisa Hensley; Peter B. Jahrling. 2018. "Testing therapeutics in cell-based assays: Factors that influence the apparent potency of drugs." PLoS ONE 13, no. 3: e0194880.

Journal article
Published: 14 December 2017 in The Journal of Infectious Diseases
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The first reported outbreak of Ebola virus disease occurred in 1976 in Yambuku, Democratic Republic of Congo. Antibody responses in survivors 11 years after infection have been documented. However, this report is the first characterization of anti-Ebola virus antibody persistence and neutralization capacity 40 years after infection. Using ELISAs we measured survivor’s immunological response to Ebola virus Zaire (EBOV) glycoprotein and nucleoprotein, and assessed VP40 reactivity. Neutralization of EBOV was measured using a pseudovirus approach and plaque reduction neutralization test with live EBOV. Some survivors from the original EBOV outbreak still harbor antibodies against all 3 measures. Interestingly, a subset of these survivors’ serum antibodies could still neutralize live virus 40 years postinitial infection. These data provide the longest documentation of both anti-Ebola serological response and neutralization capacity within any survivor cohort, extending the known duration of response from 11 years postinfection to at least 40 years after symptomatic infection.

ACS Style

Anne W Rimoin; Kai Lu; Matthew S Bramble; Imke Steffen; Reena H Doshi; Nicole A Hoff; Patrick Mukadi; Bradly P Nicholson; Vivian H Alfonso; Gerrard Olinger; Cyrus Sinai; Lauren K Yamamoto; Christina M Ramirez; Emile Okitolonda Wemakoy; Benoit Kebela Illunga; James Pettitt; James Logue; Richard S Bennett; Peter Jahrling; David L Heymann; Peter Piot; Jean Jacques Muyembe-Tamfum; Lisa E Hensley; Graham Simmons. Ebola Virus Neutralizing Antibodies Detectable in Survivors of theYambuku, Zaire Outbreak 40 Years after Infection. The Journal of Infectious Diseases 2017, 217, 223 -231.

AMA Style

Anne W Rimoin, Kai Lu, Matthew S Bramble, Imke Steffen, Reena H Doshi, Nicole A Hoff, Patrick Mukadi, Bradly P Nicholson, Vivian H Alfonso, Gerrard Olinger, Cyrus Sinai, Lauren K Yamamoto, Christina M Ramirez, Emile Okitolonda Wemakoy, Benoit Kebela Illunga, James Pettitt, James Logue, Richard S Bennett, Peter Jahrling, David L Heymann, Peter Piot, Jean Jacques Muyembe-Tamfum, Lisa E Hensley, Graham Simmons. Ebola Virus Neutralizing Antibodies Detectable in Survivors of theYambuku, Zaire Outbreak 40 Years after Infection. The Journal of Infectious Diseases. 2017; 217 (2):223-231.

Chicago/Turabian Style

Anne W Rimoin; Kai Lu; Matthew S Bramble; Imke Steffen; Reena H Doshi; Nicole A Hoff; Patrick Mukadi; Bradly P Nicholson; Vivian H Alfonso; Gerrard Olinger; Cyrus Sinai; Lauren K Yamamoto; Christina M Ramirez; Emile Okitolonda Wemakoy; Benoit Kebela Illunga; James Pettitt; James Logue; Richard S Bennett; Peter Jahrling; David L Heymann; Peter Piot; Jean Jacques Muyembe-Tamfum; Lisa E Hensley; Graham Simmons. 2017. "Ebola Virus Neutralizing Antibodies Detectable in Survivors of theYambuku, Zaire Outbreak 40 Years after Infection." The Journal of Infectious Diseases 217, no. 2: 223-231.

Journal article
Published: 19 July 2017 in Scientific Reports
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The recent epidemic of Ebola virus disease in West Africa resulted in an unprecedented number of cases and deaths. Due to the scope of the outbreak combined with the lack of available approved treatment options, there was strong motivation to investigate any potential drug which had existing data reporting anti-Ebola activity. Drugs with demonstrated antiviral activity in the nonhuman primate models already approved for another indication or for which there was existing safety data were considered to be priorities for evaluation by the World Health Organization. Sertraline hydrochloride was reported to have anti-Ebola activity in vitro alone and in combination with other approved drugs. Although the efficacy was less than 100% in the murine model, the established safety profile of this product, the potential benefit alone and in combination, as well as the lack of other available options prioritized this compound for testing in the Ebola virus intramuscular rhesus macaque challenge model. Using a blinded dosing strategy, we demonstrated that high dose sertraline monotherapy provided no benefit for the prevention of Ebola virus disease in rhesus macaques with regards to reduction of viral load, morbidity, or survival highlighting the challenges of translating results between in vitro and in vivo models.

ACS Style

Anna N. Honko; Joshua C. Johnson; Jonathan S. Marchand; Louis Huzella; Ricky D. Adams; Nicholas Oberlander; Lisa M. Torzewski; Richard S. Bennett; Lisa Hensley; Peter B. Jahrling; Gene G. Olinger. High dose sertraline monotherapy fails to protect rhesus macaques from lethal challenge with Ebola virus Makona. Scientific Reports 2017, 7, 5886 .

AMA Style

Anna N. Honko, Joshua C. Johnson, Jonathan S. Marchand, Louis Huzella, Ricky D. Adams, Nicholas Oberlander, Lisa M. Torzewski, Richard S. Bennett, Lisa Hensley, Peter B. Jahrling, Gene G. Olinger. High dose sertraline monotherapy fails to protect rhesus macaques from lethal challenge with Ebola virus Makona. Scientific Reports. 2017; 7 (1):5886.

Chicago/Turabian Style

Anna N. Honko; Joshua C. Johnson; Jonathan S. Marchand; Louis Huzella; Ricky D. Adams; Nicholas Oberlander; Lisa M. Torzewski; Richard S. Bennett; Lisa Hensley; Peter B. Jahrling; Gene G. Olinger. 2017. "High dose sertraline monotherapy fails to protect rhesus macaques from lethal challenge with Ebola virus Makona." Scientific Reports 7, no. 1: 5886.