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Gerold, Gisa
Publications (10 of 26) Show all publications
Zapatero-Belinchon, F. J., Dietzel, E., Dolnik, O., Doehner, K., Costa, R., Hertel, B., . . . von Hahn, T. (2019). Characterization of the Filovirus-Resistant Cell Line SH-SY5Y Reveals Redundant Role of Cell Surface Entry Factors. Viruses, 11(3), Article ID 275.
Open this publication in new window or tab >>Characterization of the Filovirus-Resistant Cell Line SH-SY5Y Reveals Redundant Role of Cell Surface Entry Factors
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2019 (English)In: Viruses, ISSN 1999-4915, E-ISSN 1999-4915, Vol. 11, no 3, article id 275Article in journal (Refereed) Published
Abstract [en]

Filoviruses infect a wide range of cell types with the exception of lymphocytes. The intracellular proteins cathepsin B and L, two-pore channel 1 and 2, and bona fide receptor Niemann-Pick Disease C1 (NPC1) are essential for the endosomal phase of cell entry. However, earlier steps of filoviral infection remain poorly characterized. Numerous plasma membrane proteins have been implicated in attachment but it is still unclear which ones are sufficient for productive entry. To define a minimal set of host factors required for filoviral glycoprotein-driven cell entry, we screened twelve cell lines and identified the nonlymphocytic cell line SH-SY5Y to be specifically resistant to filovirus infection. Heterokaryons of SH-SY5Y cells fused to susceptible cells were susceptible to filoviruses, indicating that SH-SY5Y cells do not express a restriction factor but lack an enabling factor critical for filovirus entry. However, all tested cell lines expressed functional intracellular factors. Global gene expression profiling of known cell surface entry factors and protein expression levels of analyzed attachment factors did not reveal any correlation between susceptibility and expression of a specific host factor. Using binding assays with recombinant filovirus glycoprotein, we identified cell attachment as the step impaired in filovirus entry in SH-SY5Y cells. Individual overexpression of attachment factors T-cell immunoglobulin and mucin domain 1 (TIM-1), Axl, Mer, or dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin (DC-SIGN) rendered SH-SY5Y cells susceptible to filovirus glycoprotein-driven transduction. Our study reveals that a lack of attachment factors limits filovirus entry and provides direct experimental support for a model of filoviral cell attachment where host factor usage at the cell surface is highly promiscuous.

Place, publisher, year, edition, pages
MDPI, 2019
Keywords
Filovirus cell entry, attachment factors redundancy, SH-SY5Y cell line, host-pathogen teractions
National Category
Microbiology in the medical area
Identifiers
urn:nbn:se:umu:diva-158595 (URN)10.3390/v11030275 (DOI)000464395500003 ()30893855 (PubMedID)
Available from: 2019-05-23 Created: 2019-05-23 Last updated: 2019-05-23Bibliographically approved
Moreno, H., Moeller, R., Fedeli, C., Gerold, G. & Kunz, S. (2019). Comparison of the Innate Immune Responses to Pathogenic and Nonpathogenic Clade B New World Arenaviruses. Journal of Virology, 93(19), Article ID e00148-19.
Open this publication in new window or tab >>Comparison of the Innate Immune Responses to Pathogenic and Nonpathogenic Clade B New World Arenaviruses
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2019 (English)In: Journal of Virology, ISSN 0022-538X, E-ISSN 1098-5514, Vol. 93, no 19, article id e00148-19Article in journal (Refereed) Published
Abstract [en]

The New World (NW) arenaviruses are a diverse group of zoonotic viruses, including several causative agents of severe hemorrhagic fevers in humans. All known human-pathogenic NW arenaviruses belong to Glade B, where they group into sublineages with phylogenetically closely related nonpathogenic viruses, e.g., the highly pathogenic Junin (JUNV) and Machupo viruses with the nonpathogenic Tacaribe virus (TCRV). Considering the close genetic relationship of nonpathogenic and pathogenic NW arenaviruses, the identification of molecular determinants of virulence is of great importance. The host cell's innate antiviral defense represents a major barrier for zoonotic infection. Here, we performed a side-by-side comparison of the innate immune responses against JUNV and TCRV in human cells. Despite similar levels of viral replication, infection with TCRV consistently induced a stronger type I interferon (IFN-I) response than JUNV infection did. Transcriptome profiling revealed upregulation of a largely overlapping set of interferon-stimulated genes in cells infected with TCRV and JUNV. Both viruses were relatively insensitive to IFN-I treatment of human cells and induced similar levels of apoptosis in the presence or absence of an IFN-I response. However, in comparison to JUNV, TCRV induced stronger activation of the innate sensor double-strand RNA-dependent protein kinase R (PKR), resulting in phosphorylation of eukaryotic translation initiation factor eIF2 alpha. Confocal microscopy studies revealed similar subcellular colocalizations of the JUNV and TCRV viral replication-transcription complexes with PKR. However, deletion of PKR by CRISPR/Cas9 hardly affected JUNV but promoted TCRV multiplication, providing the first evidence for differential innate recognition and control of pathogenic and nonpathogenic NW arenaviruses by PKR.

IMPORTANCE New World (NW) arenaviruses are a diverse family of emerging zoonotic viruses that merit significant attention as important public health problems. The close genetic relationship of nonpathogenic NW arenaviruses with their highly pathogenic cousins suggests that few mutations may be sufficient to enhance virulence. The identification of molecular determinants of virulence of NW arenaviruses is therefore of great importance. Here we undertook a side-by-side comparison of the innate immune responses against the highly pathogenic Junin virus (JUNV) and the related nonpathogenic Tacaribe virus (TCRV) in human cells. We consistently found that TCRV induces a stronger type I interferon (IFN-I) response than JUNV. Transcriptome profiling revealed an overlapping pattern of IFN-induced gene expression and similar low sensitivities to IFN-I treatment. However, the double-stranded RNA (dsRNA)-dependent protein kinase R (PKR) contributed to the control of TCRV, but not JUNV, providing the first evidence for differential innate recognition and control of JUNV and TCRV.

Place, publisher, year, edition, pages
Washington: American Society of Microbiology, 2019
Keywords
arenavirus, innate immunity, interferon, protein kinase R, zoonosis
National Category
Microbiology in the medical area
Identifiers
urn:nbn:se:umu:diva-164384 (URN)10.1128/JVI.00148-19 (DOI)000485789300001 ()31270228 (PubMedID)
Available from: 2019-11-12 Created: 2019-11-12 Last updated: 2019-11-12Bibliographically approved
Herrador, A., Fedeli, C., Radulovic, E., Campbell, K. P., Moreno, H., Gerold, G. & Kunz, S. (2019). Dynamic Dystroglycan Complexes Mediate Cell Entry of Lassa Virus. mBio, 10(2), Article ID e02869-18.
Open this publication in new window or tab >>Dynamic Dystroglycan Complexes Mediate Cell Entry of Lassa Virus
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2019 (English)In: mBio, ISSN 2161-2129, E-ISSN 2150-7511, Vol. 10, no 2, article id e02869-18Article in journal (Refereed) Published
Abstract [en]

Recognition of functional receptors by viruses is a key determinant for their host range, tissue tropism, and disease potential. The highly pathogenic Lassa virus (LASV) currently represents one of the most important emerging pathogens. The major cellular receptor for LASV in human cells is the ubiquitously expressed and evolutionary highly conserved extracellular matrix receptor dystroglycan (DG). In the host, DG interacts with many cellular proteins in a tissue-specific manner. The resulting distinct supramolecular complexes likely represent the functional units for viral entry, and preexisting protein-protein interactions may critically influence DG's function in productive viral entry. Using an unbiased shotgun proteomic approach, we define the largely unknown molecular composition of DG complexes present in highly susceptible epithelial cells that represent important targets for LASV during viral transmission. We further show that the specific composition of cellular DG complexes can affect DG's function in receptor-mediated endocytosis of the virus. Under steady-state conditions, epithelial DG complexes underwent rapid turnover via an endocytic pathway that shared some characteristics with DG-mediated LASV entry. However, compared to steady-state uptake of DG, LASV entry via DG occurred faster and critically depended on additional signaling by receptor tyrosine kinases and the downstream effector p21-activating kinase. In sum, we show that the specific molecular composition of DG complexes in susceptible cells is a determinant for productive virus entry and that the pathogen can manipulate the existing DG-linked endocytic pathway. This highlights another level of complexity of virus-receptor interaction and provides possible cellular targets for therapeutic antiviral intervention.

Importance: Recognition of cellular receptors allows emerging viruses to break species barriers and is an important determinant for their disease potential. Many virus receptors have complex tissue-specific interactomes, and preexisting protein-protein interactions may influence their function. Combining shotgun proteomics with a biochemical approach, we characterize the molecular composition of the functional receptor complexes used by the highly pathogenic Lassa virus (LASV) to invade susceptible human cells. We show that the specific composition of the receptor complexes affects productive entry of the virus, providing proof-of-concept. In uninfected cells, these functional receptor complexes undergo dynamic turnover involving an endocytic pathway that shares some characteristics with viral entry. However, steady-state receptor uptake and virus endocytosis critically differ in kinetics and underlying signaling, indicating that the pathogen can manipulate the receptor complex according to its needs. Our study highlights a remarkable complexity of LASV-receptor interaction and identifies possible targets for therapeutic antiviral intervention.

Place, publisher, year, edition, pages
American Society for Microbiology, 2019
Keywords
arenavirus, proteomics, tropism, viral entry, virus receptor
National Category
Microbiology in the medical area
Identifiers
urn:nbn:se:umu:diva-158601 (URN)10.1128/mBio.02869-18 (DOI)000465077600041 ()30914516 (PubMedID)
Available from: 2019-05-23 Created: 2019-05-23 Last updated: 2019-05-23Bibliographically approved
Fedeli, C., Torriani, G., Galan-Navarro, C., Moraz, M.-L., Moreno, H., Gerold, G. & Kunz, S. (2018). Axl can serve as entry factor for lassa virus depending on the functional glycosylation of dystroglycan. Journal of Virology, 92(5), Article ID e01613-17.
Open this publication in new window or tab >>Axl can serve as entry factor for lassa virus depending on the functional glycosylation of dystroglycan
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2018 (English)In: Journal of Virology, ISSN 0022-538X, E-ISSN 1098-5514, Vol. 92, no 5, article id e01613-17Article in journal (Refereed) Published
Abstract [en]

The highly pathogenic arenavirus Lassa virus (LASV) represents a serious public health problem in Africa. Although the principal LASV receptor, dystroglycan (DG), is ubiquitously expressed, virus binding critically depends on DG's posttranslational modification, which does not always correlate with tissue tropism. The broadly expressed phosphatidylserine receptor Axl was recently identified as an alternative LASV receptor candidate, but its role in LASV entry is unclear. Here, we investigate the exact role of Axl in LASV entry as a function of DG's posttranslational modification. We found that in the absence of functional DG, Axl can mediate LASV entry via apoptotic mimicry. Productive entry requires virus-induced receptor activation, involves macropinocytosis, and critically depends on LAMP-1. In endothelial cells that express low levels of glycosylated DG, both receptors can promote LASV entry. In sum, our study defines the roles of Axl in LASV entry and provides a rationale for targeting Axl in antiviral therapy.

Keywords
Axl, Lassa virus, attachment, dystroglycan, entry factor, macropinocytosis, receptor, viral entry
National Category
Clinical Medicine
Identifiers
urn:nbn:se:umu:diva-148138 (URN)10.1128/JVI.01613-17 (DOI)000424744800005 ()29237830 (PubMedID)
Available from: 2018-05-29 Created: 2018-05-29 Last updated: 2018-10-29Bibliographically approved
Banse, P., Moeller, R., Bruening, J., Lasswitz, L., Kahl, S., Khan, A. G., . . . Gerold, G. (2018). CD81 receptor regions outside the large extracellular loop determine hepatitis C virus entry into hepatoma cells. Viruses, 10(4), Article ID 207.
Open this publication in new window or tab >>CD81 receptor regions outside the large extracellular loop determine hepatitis C virus entry into hepatoma cells
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2018 (English)In: Viruses, ISSN 1999-4915, E-ISSN 1999-4915, Vol. 10, no 4, article id 207Article in journal (Refereed) Published
Abstract [en]

Hepatitis C virus (HCV) enters human hepatocytes using four essential entry factors, one of which is human CD81 (hCD81). The tetraspanin hCD81 contains a large extracellular loop (LEL), which interacts with the E2 glycoprotein of HCV. The role of the non-LEL regions of hCD81 (intracellular tails, four transmembrane domains, small extracellular loop and intracellular loop) is poorly understood. Here, we studied the contribution of these domains to HCV susceptibility of hepatoma cells by generating chimeras of related tetraspanins with the hCD81 LEL. Our results show that non-LEL regions in addition to the LEL determine susceptibility of cells to HCV. While closely related tetraspanins (X. tropicalis CD81 and D. rerio CD81) functionally complement hCD81 non-LEL regions, distantly related tetraspanins (C. elegans TSP9 amd D. melanogaster TSP96F) do not and tetraspanins with intermediate homology (hCD9) show an intermediate phenotype. Tetraspanin homology and susceptibility to HCV correlate positively. For some chimeras, infectivity correlates with surface expression. In contrast, the hCD9 chimera is fully surface expressed, binds HCV E2 glycoprotein but is impaired in HCV receptor function. We demonstrate that a cholesterol-coordinating glutamate residue in CD81, which hCD9 lacks, promotes HCV infection. This work highlights the hCD81 non-LEL regions as additional HCV susceptibility-determining factors.

Keywords
CD81, HCV, chimeras, cholesterol-binding residue, hepatitis C virus, receptor, susceptibility-determining domains, tetraspanin, transmembrane domain four
National Category
Infectious Medicine
Identifiers
urn:nbn:se:umu:diva-148131 (URN)10.3390/v10040207 (DOI)000435184400068 ()29677132 (PubMedID)
Available from: 2018-05-29 Created: 2018-05-29 Last updated: 2018-10-29Bibliographically approved
Lasswitz, L., Chandra, N., Arnberg, N. & Gerold, G. (2018). Glycomics and Proteomics Approaches to Investigate Early Adenovirus-Host Cell Interactions. Journal of Molecular Biology, 430(13), 1863-1882
Open this publication in new window or tab >>Glycomics and Proteomics Approaches to Investigate Early Adenovirus-Host Cell Interactions
2018 (English)In: Journal of Molecular Biology, ISSN 0022-2836, E-ISSN 1089-8638, Vol. 430, no 13, p. 1863-1882Article in journal (Refereed) Published
Abstract [en]

Adenoviruses as most viruses rely on glycan and protein interactions to attach to and enter susceptible host cells. The Adenoviridae family comprises more than 80 human types and they differ in their attachment factor and receptor usage, which likely contributes to the diverse tropism of the different types. In the past years, methods to systematically identify glycan and protein interactions have advanced. In particular sensitivity, speed and coverage of mass spectrometric analyses allow for high-throughput identification of glycans and peptides separated by liquid chromatography. Also, developments in glycan microarray technologies have led to targeted, high-throughput screening and identification of glycan-based receptors. The mapping of cell surface interactions of the diverse adenovirus types has implications for cell, tissue, and species tropism as well as drug development. Here we review known adenovirus interactions with glycan- and protein-based receptors, as well as glycomics and proteomics strategies to identify yet elusive virus receptors and attachment factors. We finally discuss challenges, bottlenecks, and future research directions in the field of non-enveloped virus entry into host cells.

Keywords
adenovirus, glycomis, host cell interactions, proteomics, virus entry
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy) Microbiology in the medical area
Identifiers
urn:nbn:se:umu:diva-148152 (URN)10.1016/j.jmb.2018.04.039 (DOI)000436224800004 ()29746851 (PubMedID)2-s2.0-85047296623 (Scopus ID)
Available from: 2018-05-29 Created: 2018-05-29 Last updated: 2018-09-28Bibliographically approved
Bruening, J., Lasswitz, L., Banse, P., Kahl, S., Marinach, C., Vondran, F. W., . . . Gerold, G. (2018). Hepatitis C virus enters liver cells using the CD81 receptor complex proteins calpain-5 and CBLB. PLoS Pathogens, 14(7), Article ID e1007111.
Open this publication in new window or tab >>Hepatitis C virus enters liver cells using the CD81 receptor complex proteins calpain-5 and CBLB
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2018 (English)In: PLoS Pathogens, ISSN 1553-7366, E-ISSN 1553-7374, Vol. 14, no 7, article id e1007111Article in journal (Refereed) Published
Abstract [en]

Hepatitis C virus (HCV) and the malaria parasite Plasmodium use the membrane protein CD81 to invade human liver cells. Here we mapped 33 host protein interactions of CD81 in primary human liver and hepatoma cells using high-resolution quantitative proteomics. In the CD81 protein network, we identified five proteins which are HCV entry factors or facilitators including epidermal growth factor receptor (EGFR). Notably, we discovered calpain-5 (CAPN5) and the ubiquitin ligase Casitas B-lineage lymphoma proto-oncogene B (CBLB) to form a complex with CD81 and support HCV entry. CAPN5 and CBLB were required for a post-binding and pre-replication step in the HCV life cycle. Knockout of CAPN5 and CBLB reduced susceptibility to all tested HCV genotypes, but not to other enveloped viruses such as vesicular stomatitis virus and human coronavirus. Furthermore, Plasmodium sporozoites relied on a distinct set of CD81 interaction partners for liver cell entry. Our findings reveal a comprehensive CD81 network in human liver cells and show that HCV and Plasmodium highjack selective CD81 interactions, including CAPN5 and CBLB for HCV, to invade cells.

National Category
Microbiology in the medical area
Identifiers
urn:nbn:se:umu:diva-150844 (URN)10.1371/journal.ppat.1007111 (DOI)000440351300009 ()30024968 (PubMedID)
Available from: 2018-08-31 Created: 2018-08-31 Last updated: 2018-08-31Bibliographically approved
Gerold, G., Bruening, J., Weigel, B. & Pietschmann, T. (2017). Protein Interactions during the Flavivirus and Hepacivirus Life Cycle. Molecular & Cellular Proteomics, 16(4), 75-91
Open this publication in new window or tab >>Protein Interactions during the Flavivirus and Hepacivirus Life Cycle
2017 (English)In: Molecular & Cellular Proteomics, ISSN 1535-9476, E-ISSN 1535-9484, Vol. 16, no 4, p. 75-91Article in journal (Refereed) Published
Abstract [en]

interaction proteomics and why we believe these challenges should be met.

Place, publisher, year, edition, pages
American Society for Biochemistry and Molecular Biology, 2017
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:umu:diva-148154 (URN)10.1074/mcp.R116.065649 (DOI)000398812800008 ()28077444 (PubMedID)
Note

Supplement: 1

Available from: 2018-05-29 Created: 2018-05-29 Last updated: 2018-10-16Bibliographically approved
Bruening, J., Weigel, B. & Gerold, G. (2017). The Role of Type III Interferons in Hepatitis C Virus Infection and Therapy. Journal of Immunology Research, Article ID 7232361.
Open this publication in new window or tab >>The Role of Type III Interferons in Hepatitis C Virus Infection and Therapy
2017 (English)In: Journal of Immunology Research, ISSN 2314-8861, E-ISSN 2314-7156, article id 7232361Article in journal (Refereed) Published
Abstract [en]

The human interferon (IFN) response is a key innate immune mechanism to fight virus infection. IFNs are host-encoded secreted proteins, which induce IFN-stimulated genes (ISGs) with antiviral properties. Among the three classes of IFNs, type III IFNs, also called IFN lambdas (IFNLs), are an essential component of the innate immune response to hepatitis C virus (HCV). In particular, human polymorphisms in IFNL gene loci correlate with hepatitis C disease progression and with treatment response. To date, the underlying mechanisms remain mostly elusive; however it seems clear that viral infection of the liver induces IFNL responses. As IFNL receptors show a more restricted tissue expression than receptors for other classes of IFNs, IFNL treatment has reduced side effects compared to the classical type I IFN treatment. In HCV therapy, however, IFNL will likely not play an important role as highly effective direct acting antivirals (DAA) exist. Here, we will review our current knowledge on IFNL gene expression, protein properties, signaling, ISG induction, and its implications on HCV infection and treatment. Finally, we will discuss the lessons learnt from the HCV and IFNL field for virus infections beyond hepatitis C.

Place, publisher, year, edition, pages
Hindawi Publishing Corporation, 2017
National Category
Immunology in the medical area
Identifiers
urn:nbn:se:umu:diva-148155 (URN)10.1155/2017/7232361 (DOI)000394872000001 ()28255563 (PubMedID)
Available from: 2018-05-29 Created: 2018-05-29 Last updated: 2018-10-16Bibliographically approved
Vieyres, G., Welsch, K., Gerold, G., Gentzsch, J., Kahl, S., Vondran, F. W., . . . Pietschmann, T. (2016). ABHD5/CGI-58, the Chanarin-Dorfman Syndrome Protein, Mobilises Lipid Stores for Hepatitis C Virus Production. PLoS Pathogens, 12(4), Article ID e1005568.
Open this publication in new window or tab >>ABHD5/CGI-58, the Chanarin-Dorfman Syndrome Protein, Mobilises Lipid Stores for Hepatitis C Virus Production
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2016 (English)In: PLoS Pathogens, ISSN 1553-7366, E-ISSN 1553-7374, Vol. 12, no 4, article id e1005568Article in journal (Refereed) Published
Abstract [en]

Hepatitis C virus (HCV) particles closely mimic human very-low-density lipoproteins (VLDL) to evade humoral immunity and to facilitate cell entry. However, the principles that govern HCV association with VLDL components are poorly defined. Using an siRNA screen, we identified ABHD5 (α/β hydrolase domain containing protein 5, also known as CGI-58) as a new host factor promoting both virus assembly and release. ABHD5 associated with lipid droplets and triggered their hydrolysis. Importantly, ABHD5 Chanarin-Dorfman syndrome mutants responsible for a rare lipid storage disorder in humans were mislocalised, and unable to consume lipid droplets or support HCV production. Additional ABHD5 mutagenesis revealed a novel tribasic motif that does not influence subcellular localization but determines both ABHD5 lipolytic and proviral properties. These results indicate that HCV taps into the lipid droplet triglyceride reservoir usurping ABHD5 lipase cofactor function. They also suggest that the resulting lipid flux, normally devoted to VLDL synthesis, also participates in the assembly and release of the HCV lipo-viro-particle. Altogether, our study provides the first association between the Chanarin-Dorfman syndrome protein and an infectious disease and sheds light on the hepatic manifestations of this rare genetic disorder as well as on HCV morphogenesis.

Place, publisher, year, edition, pages
Public Library Science, 2016
National Category
Microbiology in the medical area
Identifiers
urn:nbn:se:umu:diva-148140 (URN)10.1371/journal.ppat.1005568 (DOI)000378156900049 ()27124600 (PubMedID)
Available from: 2018-05-29 Created: 2018-05-29 Last updated: 2018-10-15Bibliographically approved
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