Comparative Transcriptomics in #Ebola #Makona-Infected #Ferrets, Nonhuman #Primates, and #Humans (J Infect Dis., abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

J Infect Dis. 2018 Nov 22;218(suppl_5):S486-S495. doi: 10.1093/infdis/jiy455.

Comparative Transcriptomics in Ebola Makona-Infected Ferrets, Nonhuman Primates, and Humans.

Cross RW1,2, Speranza E3, Borisevich V1,2, Widen SG4, Wood TG4, Shim RS5, Adams RD5, Gerhardt DM5, Bennett RS5, Honko AN5, Johnson JC5, Hensley LE5, Geisbert TW1,2, Connor JH4,3.

Author information: 1 Galveston National Laboratory, University of Texas Medical Branch, Galveston. 2 Departments of Microbiology and Immunology, University of Texas Medical Branch, Galveston. 3 Department of Microbiology, Bioinformatics Program, National Emerging Infectious Disease Laboratories, Boston University, Massachusetts. 4 Departments of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston. 5 Integrated Research Facility at Fort Detrick, Division of Clinical Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Frederick, Maryland.

 

Abstract

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.

PMID: 30476250 DOI: 10.1093/infdis/jiy455

Keywords: Ebola; Ebola-Makona; Animal models.

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Naturally-occurring single #mutations in #Ebola observably impact #infectivity (J Virol., abstract)

[Source: Journal of Virology, full page: (LINK). Abstract, edited.]

Naturally-occurring single mutations in Ebola observably impact infectivity

Gary Wong, Shihua He, Anders Leung, Wenguang Cao, Yuhai Bi, Zirui Zhang, Wenjun Zhu, Liang Wang, Yuhui Zhao, Keding Cheng, Di Liu, Wenjun Liu, Darwyn Kobasa, George F. Gao,Xiangguo Qiu

DOI: 10.1128/JVI.01098-18

 

ABSTRACT

Sequencing of Ebola virus (EBOV) genomes during the 2014-16 epidemic identified several naturally-occurring, dominant mutations potentially impacting virulence or tropism. Here, we characterized EBOV variants carrying one of the following substitutions: A82V in the glycoprotein (GP), R111C in the nucleoprotein (NP), or D759G in the RNA-dependent RNA-polymerase (L). Compared with wild-type EBOV/C07 (WT), NP and L mutants conferred a replication advantage in monkey VeroE6, human A549 and insectivorous bat Tb1.Lu cells, while L displayed a disadvantage in human Huh7 cells. GP mutant replication was significantly delayed in Tb1.Lu and similar to WT in other cells. The L mutant was less virulent, as evidenced by increased survival in mice and a significantly delayed time to death in ferrets, but increased lengths of EBOV shedding may have contributed to the prolonged epidemic. Our results show that single substitutions can have observable impacts on EBOV pathogenicity and provide a framework to study other mutations.

 

IMPORTANCE

During the Ebola virus (EBOV) disease outbreak in West Africa in 2014-16, it was discovered that several mutations in the virus emerged and became prevalent in the human population. This suggests that these mutations may play a role impacting viral fitness. We investigated three of these previously identified mutations (in the glycoprotein (GP), nucleoprotein (NP) or RNA-dependent RNA polymerase (L)) in cell culture as well as in mice and ferrets by generating recombinant viruses (based on an early West African EBOV strain) carrying one of these mutations. The NP and L mutations appear to decrease virulence, whereas GP slightly increases virulence but mainly impacts viral tropism. Our results show that these single mutations can impact EBOV virulence in animals and have implications for the rational design of efficacious antiviral therapies against these infections.

© Crown copyright 2018. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

Keywords: Ebola; Ebola-Makona.

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Comparative #Transcriptomics in #Ebola #Makona-Infected #Ferrets, Nonhuman #Primates, and #Humans (J Infect Dis., abstract)

[Source: Journal of Infectious Diseases, full page: (LINK). Abstract, edited.]

Comparative Transcriptomics in Ebola Makona-Infected Ferrets, Nonhuman Primates, and Humans

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 N Honko, Joshua C Johnson, Lisa E Hensley, Thomas W Geisbert, John H Connor

The Journal of Infectious Diseases, jiy455, https://doi.org/10.1093/infdis/jiy455

Published: 23 August 2018

 

Abstract

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.

animal model, Ebola virus, ferret, transcriptomics, vaccines

Issue Section: Supplement Article

© The Author(s) 2018. Published by Oxford University Press for the Infectious Diseases Society of America.

This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model)

Keywords: Ebola; Ebola-Makona; Animal Models.

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#Serological #Investigation of Laboratory-Confirmed and Suspected #Ebola Virus Disease #Patients During the Late Phase of the #Ebola #Outbreak in #SierraLeone (Virol Sin., abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

Virol Sin. 2018 Jul 31. doi: 10.1007/s12250-018-0044-z. [Epub ahead of print]

Serological Investigation of Laboratory-Confirmed and Suspected Ebola Virus Disease Patients During the Late Phase of the Ebola Outbreak in Sierra Leone.

Liu Y1,2, Sun Y3,2, Wu W1, Li A1, Yang X4,2, Zhang S1, Li C1, Su Q1,2, Cai S5,2, Sun D6,2, Hu H7,2, Zhang Z8,2, Yang X8,2, Kamara I2,9, Koroma S2,9, Bangura G2,9, Tia A2,9, Kamara A9, Lebby M9, Kargbo B9, Li J1, Wang S1, Dong X1, Shu Y1, Xu W1, Gao GF1,10, Wu G1, Li D1, Liu WJ11,12, Liang M13.

Author information: 1 Key Laboratory of Medical Virology and Viral Diseases, Ministry of Health, National Institute for Viral Disease Control and Prevention (IVDC), Chinese Center for Disease Control and Prevention (China CDC), Beijing, 102206, China. 2 Sierra Leone-China Friendship Biological Safety Laboratory (SLE-CHN Bio-safety Lab), Freetown, 999127, Sierra Leone. 3 Beijing Center for Disease Control and Prevention (Beijing CDC, China), Beijing, 100013, China. 4 Jilin Provincial Center for Disease Control and Prevention (Jilin CDC, China), Jilin, 130021, China. 5 Fujian Provincial Center for Disease Control and Prevention (Fujian CDC, China), Fuzhou, 350001, China. 6 Shandong Provincial Center for Disease Control and Prevention (Shandong CDC, China), Jinan, 250014, China. 7 Jiangsu Provincial Center for Disease Control and Prevention (Jiangsu CDC, China), Nanjing, 210009, China. 8 Beijing Institute of Biotechnology, Beijing, 100071, China. 9 The Ministry of Health and Sanitation (MoHS, SL), Freetown, 999127, Sierra Leone. 10 CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China. 11 Key Laboratory of Medical Virology and Viral Diseases, Ministry of Health, National Institute for Viral Disease Control and Prevention (IVDC), Chinese Center for Disease Control and Prevention (China CDC), Beijing, 102206, China. liujun@ivdc.chinacdc.cn. 12 Sierra Leone-China Friendship Biological Safety Laboratory (SLE-CHN Bio-safety Lab), Freetown, 999127, Sierra Leone. liujun@ivdc.chinacdc.cn. 13 Key Laboratory of Medical Virology and Viral Diseases, Ministry of Health, National Institute for Viral Disease Control and Prevention (IVDC), Chinese Center for Disease Control and Prevention (China CDC), Beijing, 102206, China. mifangl@163.com.

 

Abstract

This study aimed to investigate the serological characteristics of Ebola virus (EBOV) infection during the late phase of the Ebola outbreak in Sierra Leone. In total, 877 blood samples from 694 suspected Ebola virus disease (EVD) cases assessed from March to December 2015, were analyzed via real-time reverse transcription polymerase chain reaction (RT-PCR) for viral RNA and enzyme-linked immunosorbent assay (ELISA) and Luminex to detect antibodies against EBOV. Viral load and EBOV-specific IgM/IgG titers displayed a declining trend during March to December 2015. Viral RNA load decreased rapidly at earlier stages after disease onset, while EBOV-specific IgM and IgG still persisted in 58.1% (18/31) and 93.5% (29/31) of the confirmed EVD patients and in 3.8% (25/663) and 17.8% (118/663) of the RNA-negative suspected patients in the later phase, respectively. Dynamic analysis of longitudinally collected samples from eight EVD patients revealed typically reversed trends of declining viral load and increasing IgM and/or IgG titers in response to the EBOV infection. The present results indicate that certain populations of Sierra Leone developed immunity to an EBOV infection in the late phase of the outbreak, providing novel insights into the risk assessment of EBOV infections among human populations.

KEYWORDS: Ebola virus (EBOV); IgG; IgM; Late phase; Serologic investigation

PMID: 30066045 DOI: 10.1007/s12250-018-0044-z

Keywords: Ebola; Ebola-Makona; Sierra Leone; Seroprevalence.

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#Persistence of #Ebola virus after the end of widespread #transmission in #Liberia: an #outbreak #report (Lancet Infect Dis., abstract)

[Source: The Lancet Infectious Diseases, full page: (LINK). Abstract, edited.]

Persistence of Ebola virus after the end of widespread transmission in Liberia: an outbreak report

Emily Kainne Dokubo, MD*, Annika Wendland, MPH*, Suzanne E Mate, PhD*, Jason T Ladner, PhD*, Esther L Hamblion, PhD, Philomena Raftery, MSc, David J Blackley, DrPH, A Scott Laney, PhD, Nuha Mahmoud, MD, Gloria Wayne-Davies, MPH, Lisa Hensley, PhD, Eric Stavale, BSc, Lawrence Fakoli, BSc, Christopher Gregory, MD, Tai-Ho Chen, MD, Augustine Koryon, BSc, Denise Roth Allen, PhD, Jennifer Mann, MPH, Andrew Hickey, PhD, John Saindon, DrHSc, Mehboob Badini, MBBS, April Baller, MD, Peter Clement, MD, Fatorma Bolay, PhD, Yatta Wapoe, MD, Michael R Wiley, PhD, James Logue, BSc, Bonnie Dighero-Kemp, BSc, Elizabeth Higgs, MD, Alex Gasasira, MBChB, Desmond E Williams, MD, Bernice Dahn, MD, Francis Kateh, MD, Tolbert Nyenswah, MPH, Gustavo Palacios, PhD†, Mosoka P Fallah, PhD†

*Joint first authors

†Contributed equally

Published: 23 July 2018 / DOI: https://doi.org/10.1016/S1473-3099(18)30417-1

© 2018 Elsevier Ltd. All rights reserved.

 

Summary

Background

Outbreak response efforts for the 2014–15 Ebola virus disease epidemic in west Africa brought widespread transmission to an end. However, subsequent clusters of infection have occurred in the region. An Ebola virus disease cluster in Liberia in November, 2015, that was identified after a 15-year-old boy tested positive for Ebola virus infection in Monrovia, raised the possibility of transmission from a persistently infected individual.

Methods

Case investigations were done to ascertain previous contact with cases of Ebola virus disease or infection with Ebola virus. Molecular investigations on blood samples explored a potential linkage between Ebola virus isolated from cases in this November, 2015, cluster and epidemiologically linked cases from the 2014–15 west African outbreak, according to the national case database.

Findings

The cluster investigated was the family of the index case (mother, father, three siblings). Ebola virus genomes assembled from two cases in the November, 2015, cluster, and an epidemiologically linked Ebola virus disease case in July, 2014, were phylogenetically related within the LB5 sublineage that circulated in Liberia starting around August, 2014. Partial genomes from two additional individuals, one from each cluster, were also consistent with placement in the LB5 sublineage. Sequencing data indicate infection with a lineage of the virus from a former transmission chain in the country. Based on serology and epidemiological and genomic data, the most plausible scenario is that a female case in the November, 2015, cluster survived Ebola virus disease in 2014, had viral persistence or recurrent disease, and transmitted the virus to three family members a year later.

Interpretation

Investigation of the source of infection for the November, 2015, cluster provides evidence of Ebola virus persistence and highlights the risk for outbreaks after interruption of active transmission. These findings underscore the need for focused prevention efforts among survivors and sustained capacity to rapidly detect and respond to new Ebola virus disease cases to prevent recurrence of a widespread outbreak.

Funding

US Centers for Disease Control and Prevention, Defense Threat Reduction Agency, and WHO.

Keywords: Ebola; Ebola-Makona; Liberia; West Africa.

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Can #Ebola virus re-emerge from #survivors’ body #fluids other than #semen? (Lancet Infect Dis., abstract)

[Source: The Lancet Infectious Diseases, full page: (LINK). Abstract, edited.]

Can Ebola virus re-emerge from survivors’ body fluids other than semen?

Lorenzo Subissi

Published: 23 July 2018 / DOI: https://doi.org/10.1016/S1473-3099(18)30435-3

© 2018 Elsevier Ltd. All rights reserved.

 

Summary

The cohort of survivors of the 2013–16 Ebola virus disease (Ebola) epidemic in west Africa—comprising more than 17 000 people—has exceeded the total number of survivors from all previous Ebola outbreaks combined. This high number of survivors has quickly created a new research agenda in the area of filoviruses. The potential persistence of Ebola virus in body fluids was first described during the 1995 Kikwit outbreak in the Democratic Republic of Congo.1 Evidence has grown rapidly during and after the 2013–16 west African outbreak, when different studies reported Ebola virus persistence in survivors’ immunoprivileged sites.

Keywords: Ebola; Ebola-Makona; West Africa.

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Analysis of #CD8+ T cell response during the 2013–2016 #Ebola #epidemic in West #Africa (Proc Natl Acad Sci USA, abstract)

[Source: Proceedings of the National Academy of Sciences of the United States of America, full page: (LINK). Abstract, edited.]

Analysis of CD8+ T cell response during the 2013–2016 Ebola epidemic in West Africa

Saori Sakabe, Brian M. Sullivan, Jessica N. Hartnett, Refugio Robles-Sikisaka, Karthik Gangavarapu, Beatrice Cubitt, Brian C. Ware, Dylan Kotliar, Luis M. Branco, Augustine Goba, Mambu Momoh, John Demby Sandi, Lansana Kanneh, Donald S. Grant, Robert F. Garry, Kristian G. Andersen, Juan Carlos de la Torre, Pardis C. Sabeti, John S. Schieffelin, and Michael B. A. Oldstone

PNAS July 23, 2018. 201806200; published ahead of print July 23, 2018. https://doi.org/10.1073/pnas.1806200115

Contributed by Michael B. A. Oldstone, June 18, 2018 (sent for review April 13, 2018; reviewed by Arturo Casadevall and Bruce D. Walker)

 

Significance

Zaire ebolavirus (EBOV) is a viral pathogen of significant global health concern best exemplified by more than 28,000 human infections during the recent West African epidemic. Examining immunity in EBOV disease survivors has been historically difficult due to the occurrence of only small outbreaks in remote regions of central Africa. Consequently, little data exist describing EBOV-specific T cell responses during human infection. We examined virus-specific CD8+ T cell immunity in 32 Sierra Leonean survivors of the 2013–2016 epidemic. CD8+ T cells against the nucleoprotein dominated the EBOV-specific responses in this group, while a minority of individuals harbored memory CD8+ T cells against the EBOV-GP. Our data have implications in designing EBOV vaccines that can elicit cell-mediated immunity in a large group of individuals.

 

Abstract

The recent Ebola epidemic exemplified the importance of understanding and controlling emerging infections. Despite the importance of T cells in clearing virus during acute infection, little is known about Ebola-specific CD8+ T cell responses. We investigated immune responses of individuals infected with Ebola virus (EBOV) during the 2013–2016 West Africa epidemic in Sierra Leone, where the majority of the >28,000 EBOV disease (EVD) cases occurred. We examined T cell memory responses to seven of the eight Ebola proteins (GP, sGP, NP, VP24, VP30, VP35, and VP40) and associated HLA expression in survivors. Of the 30 subjects included in our analysis, CD8+ T cells from 26 survivors responded to at least one EBOV antigen. A minority, 10 of 26 responders (38%), made CD8+ T cell responses to the viral GP or sGP. In contrast, 25 of the 26 responders (96%) made response to viral NP, 77% to VP24 (20 of 26), 69% to VP40 (18 of 26), 42% (11 of 26) to VP35, with no response to VP30. Individuals making CD8+ T cells to EBOV VP24, VP35, and VP40 also made CD8+ T cells to NP, but rarely to GP. We identified 34 CD8+ T cell epitopes for Ebola. Our data indicate the immunodominance of the EBOV NP-specific T cell response and suggest that its inclusion in a vaccine along with the EBOV GP would best mimic survivor responses and help boost cell-mediated immunity during vaccination.

Ebola – virus-specific – CD8 T cells – epitopes – HLA

 

Footnotes

1 S.S. and B.M.S. contributed equally to this work.

2 To whom correspondence may be addressed. Email: bsully@scripps.edu or mbaobo@scripps.edu.

Author contributions: S.S., B.M.S., R.F.G., K.G.A., J.C.d.l.T., P.C.S., J.S.S., and M.B.A.O. designed research; S.S., B.M.S., J.N.H., R.R.-S., K.G., B.C., B.C.W., D.K., L.M.B., A.G., M.M., J.D.S., L.K., D.S.G., R.F.G., and J.S.S. performed research; S.S., B.M.S., R.R.-S., K.G., D.K., R.F.G., K.G.A., and M.B.A.O. analyzed data; and S.S., B.M.S., and M.B.A.O. wrote the paper.

Reviewers: A.C., The Johns Hopkins Bloomberg School of Public Health; and B.D.W., Ragon Institute of Massachusetts General Hospital, Massachusetts Institute of Technology, and Harvard University.

The authors declare no conflict of interest.

Data deposition: The data from this study have been deposited in the Immune Epitope Database, www.iedb.org/SubID/1000771.

This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1806200115/-/DCSupplemental.

Published under the PNAS license.

Keywords: Ebola; Ebola-Makona; Immunology; Vaccines.

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