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Article ; Online: Isolation of Primary Porcine Bronchial Epithelial Cells for Nipah Virus Infections.

Elvert, Mareike / Sauerhering, Lucie / Heiner, Anja / Maisner, Andrea

Methods in molecular biology (Clifton, N.J.)

2023 Volume 2682, Page(s) 103–120

Abstract: The Malaysian strain of Nipah virus (NiV) first emerged in 1998/99 and caused a major disease outbreak in pigs and humans. While humans developed fatal encephalitis due to a prominent infection of brain microvessels, NiV-infected pigs mostly suffered ... ...

Abstract	The Malaysian strain of Nipah virus (NiV) first emerged in 1998/99 and caused a major disease outbreak in pigs and humans. While humans developed fatal encephalitis due to a prominent infection of brain microvessels, NiV-infected pigs mostly suffered from an acute respiratory disease and efficiently spread the infection via airway secretions. To elucidate the molecular basis of the highly productive NiV replication in porcine airways in vitro, physiologically relevant cell models that have maintained functional characteristics of airway epithelia in vivo are needed. Here, we describe in detail the method of isolating bronchial epithelial cells (PBEpC) from pig lungs that can be used for NiV infection studies. After the dissection of primary bronchia and removal of the mucus and protease digestion, bronchi segments are cut open and epithelial cells are scraped off and seeded on collagen-coated cell culture flasks. With this method, it is possible to isolate about 2 × 10
MeSH term(s)	Humans ; Swine ; Animals ; Henipavirus Infections ; Epithelial Cells ; Epithelium ; Brain ; Bronchi
Language	English
Publishing date	2023-08-22
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-3283-3_8
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Cytokine Induction in Nipah Virus-Infected Primary Human and Porcine Bronchial Epithelial Cells.

Elvert, Mareike / Sauerhering, Lucie / Maisner, Andrea

The Journal of infectious diseases

2019 Volume 221, Issue Suppl 4, Page(s) S395–S400

Abstract: During the Nipah virus (NiV) outbreak in Malaysia, pigs and humans were infected. While pigs generally developed severe respiratory disease due to effective virus replication and associated inflammation processes in porcine airways, respiratory symptoms ... ...

Abstract	During the Nipah virus (NiV) outbreak in Malaysia, pigs and humans were infected. While pigs generally developed severe respiratory disease due to effective virus replication and associated inflammation processes in porcine airways, respiratory symptoms in humans were rare and less severe. To elucidate the reasons for the species-specific differences in NiV airway infections, we compared the cytokine responses as a first reaction to NiV in primary porcine and human bronchial epithelial cells (PBEpC and HBEpC, respectively). In both cell types, NiV infection resulted in the expression of type III interferons (IFN-λ). Upon infection with similar virus doses, viral RNA load and IFN expression were substantially higher in HBEpC. Even if PBEpC expressed the same viral RNA amounts as NiV-infected HBEpC, the porcine cells showed reduced IFN- and IFN-dependent antiviral gene expression. Despite this inherently limited IFN response, the expression of proinflammatory cytokines (IL-6, IL-8) in NiV-infected PBEpC was not decreased. The downregulation of antiviral activity in the presence of a functional proinflammatory cytokine response might be one of the species-specific factors contributing to efficient virus replication and acute inflammation in the lungs of pigs infected with the Malaysian NiV strain.
MeSH term(s)	Animals ; Bronchi ; Cytokines/genetics ; Cytokines/metabolism ; Epithelial Cells/virology ; Gene Expression Regulation/immunology ; Humans ; Nipah Virus/physiology ; Respiratory Mucosa/cytology ; Species Specificity ; Swine
Chemical Substances	Cytokines
Language	English
Publishing date	2019-11-24
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	3019-3
ISSN	1537-6613 ; 0022-1899
ISSN (online)	1537-6613
ISSN	0022-1899
DOI	10.1093/infdis/jiz455
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Book ; Online ; Thesis: Einfluss von Wirtsfaktoren auf die Nipahvirus-Infektion humaner und porciner Bronchial-Epithelzellen

Maisner, Andrea / Sauerhering, Lucie

2015

Title variant	Influence of host factors on Nipahvirus infection of human and porcine bronchial epithelial cells
Author's details	Lucie Sauerhering. Betreuer: Andrea Maisner
Language	German
Size	Online-Ressource
Publisher	Philipps-Universität Marburg
Publishing place	Marburg
Document type	Book ; Online ; Thesis
Thesis / German Habilitation thesis	Philipps-Universität Marburg, Diss.--Marburg, 2014
Note	Lizenzpflichtig
Database	Special collection on veterinary medicine and general parasitology

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Article ; Online: Replication of a Nipah Virus Encoding a Nuclear-Retained Matrix Protein.

Ringel, Marc / Behner, Laura / Heiner, Anja / Sauerhering, Lucie / Maisner, Andrea

The Journal of infectious diseases

2019 Volume 221, Issue Suppl 4, Page(s) S389–S394

Abstract: Nipah virus (NiV) matrix protein (NiV M) plays a major role in virus assembly. It undergoes nuclear transit before accumulating at the plasma membrane and recruiting nucleocapsids to the budding sites. Because nuclear NiV M cannot be detected in all cell ...

Abstract	Nipah virus (NiV) matrix protein (NiV M) plays a major role in virus assembly. It undergoes nuclear transit before accumulating at the plasma membrane and recruiting nucleocapsids to the budding sites. Because nuclear NiV M cannot be detected in all cell types, we wondered whether it can reach the cell surface by bypassing the nucleus. Using an M mutant with a defective nuclear export signal (MNESmut), however, we revealed that the nuclear import of M is ubiquitous, because MNESmut was retained in the nuclei of all cell types tested. Because a functional nuclear transit is a general prerequisite for M surface transport, we wanted to characterize the effect of nuclear-retained M protein in a full viral context and generated a recombinant NiV-MNESmut. Mutant NiV-MNESmut caused increased cell-cell fusion and produced lower virus titers. As expected for an assembly defective NiV, perinuclear inclusions (IBperi) were formed, but inclusions at the plasma membrane (IBPM), which probably represent the viral assembly platforms, were not found. It is interesting to note that the transport-defective MNESmut was recruited to IBperi. This probably prevents overaccumulation of nonfunctional M proteins in the cytoplasm and nuclei of NiV-infected cells and thus provides first evidence that IBperi are functionally relevant aggresome-like compartments.
MeSH term(s)	Animals ; Cell Line ; Gene Expression Regulation, Viral ; Humans ; Nipah Virus/genetics ; Nipah Virus/physiology ; Protein Transport ; Viral Matrix Proteins/genetics ; Viral Matrix Proteins/metabolism ; Viral Proteins/genetics ; Viral Proteins/metabolism ; Virus Replication/physiology
Chemical Substances	Viral Matrix Proteins ; Viral Proteins
Language	English
Publishing date	2019-11-24
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	3019-3
ISSN	1537-6613 ; 0022-1899
ISSN (online)	1537-6613
ISSN	0022-1899
DOI	10.1093/infdis/jiz440
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Article ; Online: Cyclosporin A Reveals Potent Antiviral Effects in Preclinical Models of SARS-CoV-2 Infection.

Sauerhering, Lucie / Kuznetsova, Irina / Kupke, Alexandra / Meier, Lars / Halwe, Sandro / Rohde, Cornelius / Schmidt, Jörg / Morty, Rory E / Danov, Olga / Braun, Armin / Vadász, István / Becker, Stephan / Herold, Susanne

American journal of respiratory and critical care medicine

2022 Volume 205, Issue 8, Page(s) 964–968

MeSH term(s)	Antiviral Agents/therapeutic use ; COVID-19/drug therapy ; Cyclosporine/pharmacology ; Cyclosporine/therapeutic use ; Drug Evaluation, Preclinical ; Humans ; SARS-CoV-2
Chemical Substances	Antiviral Agents ; Cyclosporine (83HN0GTJ6D)
Language	English
Publishing date	2022-04-10
Publishing country	United States
Document type	Letter ; Research Support, Non-U.S. Gov't
ZDB-ID	1180953-x
ISSN	1535-4970 ; 0003-0805 ; 1073-449X
ISSN (online)	1535-4970
ISSN	0003-0805 ; 1073-449X
DOI	10.1164/rccm.202108-1830LE
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Article: Protective CD8+ T Cell Response Induced by Modified Vaccinia Virus Ankara Delivering Ebola Virus Nucleoprotein.

Kupke, Alexandra / Volz, Asisa / Dietzel, Erik / Freudenstein, Astrid / Schmidt, Jörg / Shams-Eldin, Hosam / Jany, Sylvia / Sauerhering, Lucie / Krähling, Verena / Gellhorn Serra, Michelle / Herden, Christiane / Eickmann, Markus / Becker, Stephan / Sutter, Gerd

Vaccines

2022 Volume 10, Issue 4

Abstract: The urgent need for vaccines against Ebola virus (EBOV) was underscored by the large outbreak in West Africa (2014-2016). Since then, several promising vaccine candidates have been tested in pre-clinical and clinical studies. As a result, two vaccines ... ...

Abstract	The urgent need for vaccines against Ebola virus (EBOV) was underscored by the large outbreak in West Africa (2014-2016). Since then, several promising vaccine candidates have been tested in pre-clinical and clinical studies. As a result, two vaccines were approved for human use in 2019/2020, of which one includes a heterologous adenovirus/Modified Vaccinia virus Ankara (MVA) prime-boost regimen. Here, we tested new vaccine candidates based on the recombinant MVA vector, encoding the EBOV nucleoprotein (MVA-EBOV-NP) or glycoprotein (MVA-EBOV-GP) for their efficacy after homologous prime-boost immunization in mice. Our aim was to investigate the role of each antigen in terms of efficacy and correlates of protection. Sera of mice vaccinated with MVA-EBOV-GP were virus-neutralizing and MVA-EBOV-NP immunization readily elicited interferon-γ-producing NP-specific CD8+ T cells. While mock-vaccinated mice succumbed to EBOV infection, all vaccinated mice survived and showed drastically decreased viral loads in sera and organs. In addition, MVA-EBOV-NP vaccinated mice became susceptible to lethal EBOV infection after depletion of CD8+ T cells prior to challenge. This study highlights the potential of MVA-based vaccines to elicit humoral immune responses as well as a strong and protective CD8+ T cell response and contributes to understanding the possible underlying mechanisms.
Language	English
Publishing date	2022-03-29
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2703319-3
ISSN	2076-393X
ISSN	2076-393X
DOI	10.3390/vaccines10040533
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Article ; Online: Proteomic landscape of SARS-CoV-2- and MERS-CoV-infected primary human renal epithelial cells.

Kohli, Aneesha / Sauerhering, Lucie / Fehling, Sarah K / Klann, Kevin / Geiger, Helmut / Becker, Stephan / Koch, Benjamin / Baer, Patrick C / Strecker, Thomas / Münch, Christian

Life science alliance

2022 Volume 5, Issue 5

Abstract: Acute kidney injury is associated with mortality in COVID-19 patients. However, host cell changes underlying infection of renal cells with SARS-CoV-2 remain unknown and prevent understanding of the molecular mechanisms that may contribute to renal ... ...

Abstract	Acute kidney injury is associated with mortality in COVID-19 patients. However, host cell changes underlying infection of renal cells with SARS-CoV-2 remain unknown and prevent understanding of the molecular mechanisms that may contribute to renal pathology. Here, we carried out quantitative translatome and whole-cell proteomics analyses of primary renal proximal and distal tubular epithelial cells derived from human donors infected with SARS-CoV-2 or MERS-CoV to disseminate virus and cell type-specific changes over time. Our findings revealed shared pathways modified upon infection with both viruses, as well as SARS-CoV-2-specific host cell modulation driving key changes in innate immune activation and cellular protein quality control. Notably, MERS-CoV infection-induced specific changes in mitochondrial biology that were not observed in response to SARS-CoV-2 infection. Furthermore, we identified extensive modulation in pathways associated with kidney failure that changed in a virus- and cell type-specific manner. In summary, we provide an overview of the effects of SARS-CoV-2 or MERS-CoV infection on primary renal epithelial cells revealing key pathways that may be essential for viral replication.
MeSH term(s)	Biomarkers ; COVID-19/metabolism ; COVID-19/virology ; Cell Nucleus/genetics ; Cell Nucleus/metabolism ; Cells, Cultured ; Computational Biology/methods ; Coronavirus Infections/metabolism ; Coronavirus Infections/virology ; Epithelial Cells/metabolism ; Epithelial Cells/virology ; Gene Expression Regulation ; Host-Pathogen Interactions/genetics ; Host-Pathogen Interactions/immunology ; Humans ; Kidney ; Kidney Tubules, Distal ; Kidney Tubules, Proximal ; Middle East Respiratory Syndrome Coronavirus/physiology ; Mitochondria/genetics ; Mitochondria/metabolism ; Primary Cell Culture ; Proteome ; Proteomics/methods ; SARS-CoV-2/physiology ; Virus Replication
Chemical Substances	Biomarkers ; Proteome
Language	English
Publishing date	2022-02-02
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	2575-1077
ISSN (online)	2575-1077
DOI	10.26508/lsa.202201371
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Crystal structure of SARS-CoV-2 main protease provides a basis for design of improved α-ketoamide inhibitors.

Zhang, Linlin / Lin, Daizong / Sun, Xinyuanyuan / Curth, Ute / Drosten, Christian / Sauerhering, Lucie / Becker, Stephan / Rox, Katharina / Hilgenfeld, Rolf

Science (New York, N.Y.)

2020 Volume 368, Issue 6489, Page(s) 409–412

Abstract: The coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2) is a global health emergency. An attractive drug target among coronaviruses is the main protease ( ... ...

Abstract	The coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2) is a global health emergency. An attractive drug target among coronaviruses is the main protease (M
MeSH term(s)	Amides/chemistry ; Amides/metabolism ; Amides/pharmacology ; Animals ; Antiviral Agents/chemistry ; Antiviral Agents/metabolism ; Antiviral Agents/pharmacokinetics ; Antiviral Agents/pharmacology ; Betacoronavirus/drug effects ; Betacoronavirus/enzymology ; Binding Sites ; Cell Line, Tumor ; Coronavirus 3C Proteases ; Crystallography, X-Ray ; Cysteine Endopeptidases/chemistry ; Cysteine Endopeptidases/metabolism ; Drug Design ; Half-Life ; Humans ; Lung/metabolism ; Mice ; Models, Molecular ; Protease Inhibitors/chemistry ; Protease Inhibitors/metabolism ; Protease Inhibitors/pharmacokinetics ; Protease Inhibitors/pharmacology ; Protein Domains ; Protein Multimerization ; Pyridones/chemistry ; SARS-CoV-2 ; Viral Nonstructural Proteins/chemistry ; Viral Nonstructural Proteins/metabolism ; Virus Replication/drug effects
Chemical Substances	Amides ; Antiviral Agents ; Protease Inhibitors ; Pyridones ; Viral Nonstructural Proteins ; Cysteine Endopeptidases (EC 3.4.22.-) ; Coronavirus 3C Proteases (EC 3.4.22.28)
Keywords	covid19
Language	English
Publishing date	2020-03-20
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	128410-1
ISSN	1095-9203 ; 0036-8075
ISSN (online)	1095-9203
ISSN	0036-8075
DOI	10.1126/science.abb3405
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Article ; Online: Diastereomeric Resolution Yields Highly Potent Inhibitor of SARS-CoV-2 Main Protease.

Cooper, Mark S / Zhang, Linlin / Ibrahim, Mohamed / Zhang, Kaixuan / Sun, Xinyuanyuan / Röske, Judith / Göhl, Matthias / Brönstrup, Mark / Cowell, Justin K / Sauerhering, Lucie / Becker, Stephan / Vangeel, Laura / Jochmans, Dirk / Neyts, Johan / Rox, Katharina / Marsh, Graham P / Maple, Hannah J / Hilgenfeld, Rolf

Journal of medicinal chemistry

2022 Volume 65, Issue 19, Page(s) 13328–13342

Abstract: SARS-CoV-2 is the causative agent behind the COVID-19 pandemic. The main protease ( ... ...

Abstract	SARS-CoV-2 is the causative agent behind the COVID-19 pandemic. The main protease (M
MeSH term(s)	Humans ; Antiviral Agents/chemistry ; Antiviral Agents/pharmacology ; Coronavirus 3C Proteases ; COVID-19 Drug Treatment ; Cysteine Endopeptidases/metabolism ; Pandemics ; Polyproteins ; Protease Inhibitors/chemistry ; Protease Inhibitors/pharmacology ; RNA, Viral ; SARS-CoV-2 ; Viral Nonstructural Proteins/metabolism
Chemical Substances	3C-like proteinase, SARS-CoV-2 (EC 3.4.22.-) ; Antiviral Agents ; Coronavirus 3C Proteases (EC 3.4.22.28) ; Cysteine Endopeptidases (EC 3.4.22.-) ; Polyproteins ; Protease Inhibitors ; RNA, Viral ; Viral Nonstructural Proteins ; SARS-CoV-2-IN-1
Language	English
Publishing date	2022-09-30
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	218133-2
ISSN	1520-4804 ; 0022-2623
ISSN (online)	1520-4804
ISSN	0022-2623
DOI	10.1021/acs.jmedchem.2c01131
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Article ; Online: Self-amplifying RNA vaccine protects mice against lethal Ebola virus infection.

Krähling, Verena / Erbar, Stephanie / Kupke, Alexandra / Nogueira, Sara S / Walzer, Kerstin C / Berger, Hendrik / Dietzel, Erik / Halwe, Sandro / Rohde, Cornelius / Sauerhering, Lucie / Aragão-Santiago, Letícia / Moreno Herrero, Jorge / Witzel, Sonja / Haas, Heinrich / Becker, Stephan / Sahin, Ugur

Molecular therapy : the journal of the American Society of Gene Therapy

2022 Volume 31, Issue 2, Page(s) 374–386

Abstract: Emerging and re-emerging viruses, such as Zaire Ebola virus (EBOV), pose a global threat and require immediate countermeasures, including the rapid development of effective vaccines that are easy to manufacture. Synthetic self-amplifying RNAs (saRNAs) ... ...

Abstract	Emerging and re-emerging viruses, such as Zaire Ebola virus (EBOV), pose a global threat and require immediate countermeasures, including the rapid development of effective vaccines that are easy to manufacture. Synthetic self-amplifying RNAs (saRNAs) attend to these needs, being safe and strong immune stimulators that can be inexpensively produced in large quantities, using cell-free systems and good manufacturing practice. Here, the first goal was to develop and optimize an anti-EBOV saRNA-based vaccine in terms of its antigen composition and route of administration. Vaccinating mice with saRNAs expressing the EBOV glycoprotein (GP) alone or in combination with the nucleoprotein (NP) elicited antigen-specific immune responses. GP-specific antibodies showed neutralizing activity against EBOV. Strong CD4
MeSH term(s)	Animals ; Mice ; Hemorrhagic Fever, Ebola/prevention & control ; Antibodies, Viral ; Antibodies, Neutralizing ; Ebolavirus/genetics ; Glycoproteins/genetics ; Ebola Vaccines/genetics
Chemical Substances	Antibodies, Viral ; Antibodies, Neutralizing ; Glycoproteins ; Ebola Vaccines
Language	English
Publishing date	2022-10-27
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2010592-7
ISSN	1525-0024 ; 1525-0016
ISSN (online)	1525-0024
ISSN	1525-0016
DOI	10.1016/j.ymthe.2022.10.011
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