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PML nuclear bodies (PML-NBs) are dynamic interchromosomal macromolecular complexes implicated in epigenetic regulation as well as antiviral defense. During herpesvirus infection, PML-NBs induce epigenetic silencing of viral genomes, however, this defense is antagonized by viral regulatory proteins such as IE1 of human cytomegalovirus (HCMV). Here, we show that PML-NBs undergo a drastic rearrangement into highly enlarged PML cages upon infection with IE1-deficient HCMV. Importantly, our results demonstrate that dual signaling by interferon and DNA damage response is required to elicit giant PML-NBs. DNA labeling revealed that invading HCMV genomes are entrapped inside PML-NBs and remain stably associated with PML cages in a transcriptionally repressed state. Intriguingly, by correlative light and transmission electron microscopy (EM), we observed that PML cages also entrap newly assembled viral capsids demonstrating a second defense layer in cells with incomplete first line response. Further characterization by 3D EM showed that hundreds of viral capsids are tightly packed into several layers of fibrous PML. Overall, our data indicate that giant PML-NBs arise via combined interferon and DNA damage signaling which triggers entrapment of both nucleic acids and proteinaceous components. This represents a multilayered defense strategy to act in a cytoprotective manner and to combat viral infections.
Myriam Scherer; Clarissa Read; Gregor Neusser; Christine Kranz; Regina Müller; Florian Full; Sonja Wörz; Anna Reichel; Eva-Maria Schilling; Paul Walther; Thomas Stamminger. Dual signaling via interferon and DNA damage response elicits entrapment by giant PML nuclear bodies. 2021, 1 .
AMA StyleMyriam Scherer, Clarissa Read, Gregor Neusser, Christine Kranz, Regina Müller, Florian Full, Sonja Wörz, Anna Reichel, Eva-Maria Schilling, Paul Walther, Thomas Stamminger. Dual signaling via interferon and DNA damage response elicits entrapment by giant PML nuclear bodies. . 2021; ():1.
Chicago/Turabian StyleMyriam Scherer; Clarissa Read; Gregor Neusser; Christine Kranz; Regina Müller; Florian Full; Sonja Wörz; Anna Reichel; Eva-Maria Schilling; Paul Walther; Thomas Stamminger. 2021. "Dual signaling via interferon and DNA damage response elicits entrapment by giant PML nuclear bodies." , no. : 1.
Restriction factors are potent antiviral proteins that constitute a first line of intracellular defense by blocking viral replication and spread. During co-evolution, however, viruses have developed antagonistic proteins to modulate or degrade the restriction factors of their host. To ensure the success of lytic replication, the herpesvirus human cytomegalovirus (HCMV) expresses the immediate-early protein IE1, which acts as an antagonist of antiviral, subnuclear structures termed PML nuclear bodies (PML-NBs). IE1 interacts directly with PML, the key protein of PML-NBs, through its core domain and disrupts the dot-like multiprotein complexes thereby abrogating the antiviral effects. Here we present the crystal structures of the human and rat cytomegalovirus core domain (IE1CORE). We found that IE1CORE domains, also including the previously characterized IE1CORE of rhesus CMV, form a distinct class of proteins that are characterized by a highly similar and unique tertiary fold and quaternary assembly. This contrasts to a marked amino acid sequence diversity suggesting that strong positive selection evolved a conserved fold, while immune selection pressure may have fostered sequence divergence of IE1. At the same time, we detected specific differences in the helix arrangements of primate versus rodent IE1CORE structures. Functional characterization revealed a conserved mechanism of PML-NB disruption, however, primate and rodent IE1 proteins were only effective in cells of the natural host species but not during cross-species infection. Remarkably, we observed that expression of HCMV IE1 allows rat cytomegalovirus replication in human cells. We conclude that cytomegaloviruses have evolved a distinct protein tertiary structure of IE1 to effectively bind and inactivate an important cellular restriction factor. Furthermore, our data show that the IE1 fold has been adapted to maximize the efficacy of PML targeting in a species-specific manner and support the concept that the PML-NBs-based intrinsic defense constitutes a barrier to cross-species transmission of HCMV.
Johannes Schweininger; Myriam Scherer; Franziska Rothemund; Eva-Maria Schilling; Sonja Wörz; Thomas Stamminger; Yves A. Muller. Cytomegalovirus immediate-early 1 proteins form a structurally distinct protein class with adaptations determining cross-species barriers. PLOS Pathogens 2021, 17, 1 .
AMA StyleJohannes Schweininger, Myriam Scherer, Franziska Rothemund, Eva-Maria Schilling, Sonja Wörz, Thomas Stamminger, Yves A. Muller. Cytomegalovirus immediate-early 1 proteins form a structurally distinct protein class with adaptations determining cross-species barriers. PLOS Pathogens. 2021; 17 (8):1.
Chicago/Turabian StyleJohannes Schweininger; Myriam Scherer; Franziska Rothemund; Eva-Maria Schilling; Sonja Wörz; Thomas Stamminger; Yves A. Muller. 2021. "Cytomegalovirus immediate-early 1 proteins form a structurally distinct protein class with adaptations determining cross-species barriers." PLOS Pathogens 17, no. 8: 1.
After emerging in Wuhan, China, in December 2019, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has quickly spread across the globe. A pandemic was declared by the World Health Organization on March 11, 2020. By July 22, 2020, more than 15 million infections had been detected worldwide and the novel coronavirus disease 2019 (COVID-19) had claimed more than 600 000 lives.1 Epidemiological data from the early stage of the pandemic suggested that children show a much milder disease course than adults.2-6 In addition, it matches the epidemiological features of the 2002-2003 SARS pandemic.7 Overall, children younger than 18 years are thought to account for only 1% to 2% of detected COVID-19 cases worldwide.8
Burkhard Tönshoff; Barbara Müller; Roland Elling; Hanna Renk; Peter Meissner; Hartmut Hengel; Sven F. Garbade; Meinhard Kieser; Kathrin Jeltsch; Jürgen Grulich-Henn; Julia Euler; Maximilian Stich; Kristine Chobanyan-Jürgens; Maria Zernickel; Aleš Janda; Lena Wölfle; Thomas Stamminger; Thomas Iftner; Tina Ganzenmueller; Christian Schmitt; Tessa Görne; Vibor Laketa; Sylvia Olberg; Anna Plaszczyca; Mirko Cortese; Ralf Bartenschlager; Constantin Pape; Roman Remme; Daniela Huzly; Marcus Panning; Sebastian Weigang; Sebastian Giese; Kevin Ciminski; Jakob Ankerhold; Georg Kochs; Martin Schwemmle; Rupert Handgretinger; Charlotte M. Niemeyer; Corinna Engel; Winfried V. Kern; Georg Friedrich Hoffmann; Axel R. Franz; Philipp Henneke; Klaus-Michael Debatin; Hans-Georg Kräusslich. Prevalence of SARS-CoV-2 Infection in Children and Their Parents in Southwest Germany. JAMA Pediatrics 2021, 175, 586 .
AMA StyleBurkhard Tönshoff, Barbara Müller, Roland Elling, Hanna Renk, Peter Meissner, Hartmut Hengel, Sven F. Garbade, Meinhard Kieser, Kathrin Jeltsch, Jürgen Grulich-Henn, Julia Euler, Maximilian Stich, Kristine Chobanyan-Jürgens, Maria Zernickel, Aleš Janda, Lena Wölfle, Thomas Stamminger, Thomas Iftner, Tina Ganzenmueller, Christian Schmitt, Tessa Görne, Vibor Laketa, Sylvia Olberg, Anna Plaszczyca, Mirko Cortese, Ralf Bartenschlager, Constantin Pape, Roman Remme, Daniela Huzly, Marcus Panning, Sebastian Weigang, Sebastian Giese, Kevin Ciminski, Jakob Ankerhold, Georg Kochs, Martin Schwemmle, Rupert Handgretinger, Charlotte M. Niemeyer, Corinna Engel, Winfried V. Kern, Georg Friedrich Hoffmann, Axel R. Franz, Philipp Henneke, Klaus-Michael Debatin, Hans-Georg Kräusslich. Prevalence of SARS-CoV-2 Infection in Children and Their Parents in Southwest Germany. JAMA Pediatrics. 2021; 175 (6):586.
Chicago/Turabian StyleBurkhard Tönshoff; Barbara Müller; Roland Elling; Hanna Renk; Peter Meissner; Hartmut Hengel; Sven F. Garbade; Meinhard Kieser; Kathrin Jeltsch; Jürgen Grulich-Henn; Julia Euler; Maximilian Stich; Kristine Chobanyan-Jürgens; Maria Zernickel; Aleš Janda; Lena Wölfle; Thomas Stamminger; Thomas Iftner; Tina Ganzenmueller; Christian Schmitt; Tessa Görne; Vibor Laketa; Sylvia Olberg; Anna Plaszczyca; Mirko Cortese; Ralf Bartenschlager; Constantin Pape; Roman Remme; Daniela Huzly; Marcus Panning; Sebastian Weigang; Sebastian Giese; Kevin Ciminski; Jakob Ankerhold; Georg Kochs; Martin Schwemmle; Rupert Handgretinger; Charlotte M. Niemeyer; Corinna Engel; Winfried V. Kern; Georg Friedrich Hoffmann; Axel R. Franz; Philipp Henneke; Klaus-Michael Debatin; Hans-Georg Kräusslich. 2021. "Prevalence of SARS-CoV-2 Infection in Children and Their Parents in Southwest Germany." JAMA Pediatrics 175, no. 6: 586.
Cytomegalovirus (CMV) infection is a risk factor for bronchiolitis obliterans (BO), one form of chronic lung allograft dysfunction (CLAD). The viral chemokine receptor M33 is essential for successful spread of murine CMV to host salivary glands. In the present study we investigated the impact of M33 on chronic airway rejection. MHC I-mismatched tracheas of C·B10-H2b/LilMcdJ mice were transplanted into BALB/c (H2d) recipients and infected at different dates with wild type (WT) or M33-deleted (delM33) MCMV representing clinical settings of viral recipient (R)-donor (D)-serostatus: (D−/R+) or (D+/R-). Grafts were recovered for gene expression and histological / immunofluorescence analysis, respectively. Evaluations showed significantly increased signs of chronic rejection in WT-infected mice compared to uninfected allografts seen in lower epithelium/lamina propria-ratio (ELR) (ELR 0.46 ± 0.07 [WT post] vs. ELR 0.66 ± 0.10 [non-inf.]; p < 0.05). The rejection in delM33-infected groups was significantly reduced vs. WT-infected groups (0.67 ± 0.04 [delM33 post]; vs. WT post p < 0.05). Furthermore, decreased rejection was observed in WT pre-infected compared to post-infected groups (0.56 ± 0.08 [WT pre]; vs. WT post p < 0.05). CD8+ T cell infiltration was significantly higher in WT-post compared to the delM33 infected or non-infected allografts. These data support the role of the CMV in accelerating CLAD. The deletion of chemokine receptor M33 leads to attenuated rejection.
Isabella Hanka; Thomas Stamminger; Martina Ramsperger-Gleixner; Annika V. Kuckhahn; Regina Müller; Michael Weyand; Christian Heim. Role of CMV chemokine receptor M33 in airway graft rejection in a mouse transplant model. Transplant Immunology 2021, 67, 101415 .
AMA StyleIsabella Hanka, Thomas Stamminger, Martina Ramsperger-Gleixner, Annika V. Kuckhahn, Regina Müller, Michael Weyand, Christian Heim. Role of CMV chemokine receptor M33 in airway graft rejection in a mouse transplant model. Transplant Immunology. 2021; 67 ():101415.
Chicago/Turabian StyleIsabella Hanka; Thomas Stamminger; Martina Ramsperger-Gleixner; Annika V. Kuckhahn; Regina Müller; Michael Weyand; Christian Heim. 2021. "Role of CMV chemokine receptor M33 in airway graft rejection in a mouse transplant model." Transplant Immunology 67, no. : 101415.
A growing body of evidence addressing the involvement of human cytomegalovirus (HCMV) in malignancies had directed attention to the oncomodulation paradigm. HCMV-DB infected human mammary epithelial cells (HMECs) in culture showed the emergence of clusters of rapidly proliferating, spheroid-shaped transformed cells named CTH (CMV-Transformed HMECs) cells. CTH cells assessment suggests a direct contribution of HCMV to oncogenesis, from key latent and lytic genes activating oncogenic pathways to fueling tumor evolution. We hypothesized that the presence of HCMV genome in CTH cells is of pivotal importance for determining its oncogenic potential. We previously reported the detection of a long non-coding (lnc) RNA4.9 gene in CTH cells. Therefore, we assessed here the presence of UL69 gene, located nearby and downstream of the lncRNA4.9 gene, in CTH cells. The HCMV UL69 gene in CTH cells was detected using polymerase chain reaction (PCR) and sequencing of UL69 gene was performed using Sanger method. The corresponding amino acid sequence was then blasted against the UL69 sequence derived from HCMV-DB genome using NCBI Protein BLAST tool. A 99% identity was present between the nucleotide sequence present in CTH cells and HCMV-DB genome. UL69 transcript was detected in RNA extracts of CTH cells, using a reverse transcription polymerase chain reaction (RT-PCR) assay, and pUL69 protein was identified in CTH lysates using western blotting. Ganciclovir-treated CTH cells showed a decrease in UL69 gene detection and cellular proliferation. In CTH cells, the knockdown of UL69 with siRNA was assessed by RT-qPCR and western blot to reveal the impact of pUL69 on HCMV replication and CTH cell proliferation. Finally, UL69 gene was detected in breast cancer biopsies. Our results indicate a close link between the UL69 gene detected in the HCMV-DB isolate used to infect HMECs, and the UL69 gene present in transformed CTH cells and tumor biopsies, further highlighting a direct role for HCMV in breast tumor development.
Sandy Haidar Ahmad; Fatima Al Moussawi; Ranim El Baba; Zeina Nehme; Sébastien Pasquereau; Amit Kumar; Chloé Molimard; Franck Monnien; Marie-Paule Algros; Racha Karaky; Thomas Stamminger; Mona Diab Assaf; Georges Herbein. Identification of UL69 Gene and Protein in Cytomegalovirus-Transformed Human Mammary Epithelial Cells. Frontiers in Oncology 2021, 11, 1 .
AMA StyleSandy Haidar Ahmad, Fatima Al Moussawi, Ranim El Baba, Zeina Nehme, Sébastien Pasquereau, Amit Kumar, Chloé Molimard, Franck Monnien, Marie-Paule Algros, Racha Karaky, Thomas Stamminger, Mona Diab Assaf, Georges Herbein. Identification of UL69 Gene and Protein in Cytomegalovirus-Transformed Human Mammary Epithelial Cells. Frontiers in Oncology. 2021; 11 ():1.
Chicago/Turabian StyleSandy Haidar Ahmad; Fatima Al Moussawi; Ranim El Baba; Zeina Nehme; Sébastien Pasquereau; Amit Kumar; Chloé Molimard; Franck Monnien; Marie-Paule Algros; Racha Karaky; Thomas Stamminger; Mona Diab Assaf; Georges Herbein. 2021. "Identification of UL69 Gene and Protein in Cytomegalovirus-Transformed Human Mammary Epithelial Cells." Frontiers in Oncology 11, no. : 1.
Flap endonuclease 1 (FEN1) is a member of the family of structure-specific endonucleases implicated in regulation of DNA damage response and DNA replication. So far, knowledge on the role of FEN1 during viral infections is limited. Previous publications indicated that poxviruses encode a conserved protein that acts in a manner similar to FEN1 to stimulate homologous recombination, double-strand break (DSB) repair and full-size genome formation. Only recently, cellular FEN1 has been identified as a key component for hepatitis B virus cccDNA formation. Here, we report on a novel functional interaction between Flap endonuclease 1 (FEN1) and the human cytomegalovirus (HCMV) immediate early protein 1 (IE1). Our results provide evidence that IE1 manipulates FEN1 in an unprecedented manner: we observed that direct IE1 binding does not only enhance FEN1 protein stability but also phosphorylation at serine 187. This correlates with nucleolar exclusion of FEN1 stimulating its DSB-generating gap endonuclease activity. Depletion of FEN1 and inhibition of its enzymatic activity during HCMV infection significantly reduced nascent viral DNA synthesis demonstrating a supportive role for efficient HCMV DNA replication. Furthermore, our results indicate that FEN1 is required for the formation of DSBs during HCMV infection suggesting that IE1 acts as viral activator of FEN1 in order to re-initiate stalled replication forks. In summary, we propose a novel mechanism of viral FEN1 activation to overcome replication fork barriers at difficult-to-replicate sites in viral genomes.
Eva-Maria Schilling; Myriam Scherer; Franziska Rothemund; Thomas Stamminger. Functional regulation of the structure-specific endonuclease FEN1 by the human cytomegalovirus protein IE1 suggests a role for the re-initiation of stalled viral replication forks. PLOS Pathogens 2021, 17, e1009460 .
AMA StyleEva-Maria Schilling, Myriam Scherer, Franziska Rothemund, Thomas Stamminger. Functional regulation of the structure-specific endonuclease FEN1 by the human cytomegalovirus protein IE1 suggests a role for the re-initiation of stalled viral replication forks. PLOS Pathogens. 2021; 17 (3):e1009460.
Chicago/Turabian StyleEva-Maria Schilling; Myriam Scherer; Franziska Rothemund; Thomas Stamminger. 2021. "Functional regulation of the structure-specific endonuclease FEN1 by the human cytomegalovirus protein IE1 suggests a role for the re-initiation of stalled viral replication forks." PLOS Pathogens 17, no. 3: e1009460.
The catabolic program of autophagy represents a powerful immune defense against viruses that is, however, counteracted by antagonizing viral factors. Understanding the exact interplay between autophagy and HCMV infection is of major importance since autophagy-related proteins emerged as promising targets for pharmacologic intervention.
Patrick König; Adriana Svrlanska; Clarissa Read; Sabine Feichtinger; Thomas Stamminger. The Autophagy-Initiating Protein Kinase ULK1 Phosphorylates Human Cytomegalovirus Tegument Protein pp28 and Regulates Efficient Virus Release. Journal of Virology 2021, 95, 1 .
AMA StylePatrick König, Adriana Svrlanska, Clarissa Read, Sabine Feichtinger, Thomas Stamminger. The Autophagy-Initiating Protein Kinase ULK1 Phosphorylates Human Cytomegalovirus Tegument Protein pp28 and Regulates Efficient Virus Release. Journal of Virology. 2021; 95 (6):1.
Chicago/Turabian StylePatrick König; Adriana Svrlanska; Clarissa Read; Sabine Feichtinger; Thomas Stamminger. 2021. "The Autophagy-Initiating Protein Kinase ULK1 Phosphorylates Human Cytomegalovirus Tegument Protein pp28 and Regulates Efficient Virus Release." Journal of Virology 95, no. 6: 1.
Human fibroblasts represent the most extensively used cell type for the investigation of lytic human cytomegalovirus (HCMV) replication. However, analyzing the function of specific proteins during infection can be challenging since primary cells are difficult to transfect. An alternative approach is the use of lentiviral transduction with vectors for stable or inducible shRNA expression. This approach provides a versatile tool to study the role of host cell factors during HCMV infection. The essential steps to achieve an efficient target protein knockdown are shRNA design, cloning, generation of transgenic lentiviral particles, and, finally, transduction of the cells. However, these steps are highly dependent on the selected vector system. Here we focus on two different vector systems and describe how to successfully generate stable and inducible knockdown fibroblasts. Additionally, we demonstrate different methods to validate the knockdown of the target protein.
Anne-Charlotte Stilp; Patrick König; Myriam Scherer; Thomas Stamminger. Stable and Inducible Gene Knockdown in Primary Human Fibroblasts: A Versatile Tool to Study the Role of Human Cytomegalovirus Host Cell Factors. Methods in Molecular Biology 2021, 2244, 115 -132.
AMA StyleAnne-Charlotte Stilp, Patrick König, Myriam Scherer, Thomas Stamminger. Stable and Inducible Gene Knockdown in Primary Human Fibroblasts: A Versatile Tool to Study the Role of Human Cytomegalovirus Host Cell Factors. Methods in Molecular Biology. 2021; 2244 ():115-132.
Chicago/Turabian StyleAnne-Charlotte Stilp; Patrick König; Myriam Scherer; Thomas Stamminger. 2021. "Stable and Inducible Gene Knockdown in Primary Human Fibroblasts: A Versatile Tool to Study the Role of Human Cytomegalovirus Host Cell Factors." Methods in Molecular Biology 2244, no. : 115-132.
Cellular restriction factors (RFs) act as important constitutive innate immune barriers against viruses. In 2006, the promyelocytic leukemia protein was described as the first RF against human cytomegalovirus (HCMV) infection which is antagonized by the viral immediate early protein IE1. Since then, at least 15 additional RFs against HCMV have been identified, including the chromatin regulatory protein SPOC1, the cytidine deaminase APOBEC3A and the dNTP triphosphohydrolase SAMHD1. These RFs affect distinct steps of the viral replication cycle such as viral entry, gene expression, the synthesis of progeny DNA or egress. This review summarizes our current knowledge on intrinsic immune mechanisms restricting HCMV replication as well as on the viral strategies to counteract the inhibitory effects of RFs. Detailed knowledge on the interplay between host RFs and antagonizing viral factors will be fundamental to develop new approaches to combat HCMV infection.
Eva-Maria Schilling; Myriam Scherer; Thomas Stamminger. Intrinsic Immune Mechanisms Restricting Human Cytomegalovirus Replication. Viruses 2021, 13, 179 .
AMA StyleEva-Maria Schilling, Myriam Scherer, Thomas Stamminger. Intrinsic Immune Mechanisms Restricting Human Cytomegalovirus Replication. Viruses. 2021; 13 (2):179.
Chicago/Turabian StyleEva-Maria Schilling; Myriam Scherer; Thomas Stamminger. 2021. "Intrinsic Immune Mechanisms Restricting Human Cytomegalovirus Replication." Viruses 13, no. 2: 179.
Nuclear egress is a common herpesviral process regulating nucleocytoplasmic capsid release. For human cytomegalovirus (HCMV), the nuclear egress complex (NEC) is determined by the pUL50-pUL53 core that regulates multicomponent assembly with NEC-associated proteins and capsids. Recently, NEC crystal structures were resolved for α-, β- and γ-herpesviruses, revealing profound structural conservation, which was not mirrored, however, by primary sequence and binding properties. The NEC binding principle is based on hook-into-groove interaction through an N-terminal hook-like pUL53 protrusion that embraces an α-helical pUL50 binding groove. So far, pUL50 has been considered as the major kinase-interacting determinant and massive phosphorylation of pUL50-pUL53 was assigned to NEC formation and functionality. Here, we addressed the question of phenotypical changes of ORF-UL50-mutated HCMVs. Surprisingly, our analyses did not detect a predominant replication defect for most of these viral mutants, concerning parameters of replication kinetics (qPCR), viral protein production (Western blot/CoIP) and capsid egress (confocal imaging/EM). Specifically, only the ORF-UL50 deletion rescue virus showed a block of genome synthesis during late stages of infection, whereas all phosphosite mutants exhibited marginal differences compared to wild-type or revertants. These results (i) emphasize a rate-limiting function of pUL50 for nuclear egress, and (ii) demonstrate that mutations in all mapped pUL50 phosphosites may be largely compensated. A refined mechanistic concept points to a multifaceted nuclear egress regulation, for which the dependence on the expression and phosphorylation of pUL50 is discussed.
Sigrun Häge; Eric Sonntag; Adriana Svrlanska; Eva Maria Borst; Anne-Charlotte Stilp; Deborah Horsch; Regina Müller; Barbara Kropff; Jens Milbradt; Thomas Stamminger; Ursula Schlötzer-Schrehardt; Manfred Marschall. Phenotypical Characterization of the Nuclear Egress of Recombinant Cytomegaloviruses Reveals Defective Replication upon ORF-UL50 Deletion but Not pUL50 Phosphosite Mutation. Viruses 2021, 13, 165 .
AMA StyleSigrun Häge, Eric Sonntag, Adriana Svrlanska, Eva Maria Borst, Anne-Charlotte Stilp, Deborah Horsch, Regina Müller, Barbara Kropff, Jens Milbradt, Thomas Stamminger, Ursula Schlötzer-Schrehardt, Manfred Marschall. Phenotypical Characterization of the Nuclear Egress of Recombinant Cytomegaloviruses Reveals Defective Replication upon ORF-UL50 Deletion but Not pUL50 Phosphosite Mutation. Viruses. 2021; 13 (2):165.
Chicago/Turabian StyleSigrun Häge; Eric Sonntag; Adriana Svrlanska; Eva Maria Borst; Anne-Charlotte Stilp; Deborah Horsch; Regina Müller; Barbara Kropff; Jens Milbradt; Thomas Stamminger; Ursula Schlötzer-Schrehardt; Manfred Marschall. 2021. "Phenotypical Characterization of the Nuclear Egress of Recombinant Cytomegaloviruses Reveals Defective Replication upon ORF-UL50 Deletion but Not pUL50 Phosphosite Mutation." Viruses 13, no. 2: 165.
Numerous studies suggest that cytomegalovirus (CMV) infection may act as isolated risk factor in the development of cardiac allograft vasculopathy (CAV). Viral G protein-coupled receptors (GPCRs) are thought to contribute to the pathogenic changes associated with CMV infection. The aim of this study was to investigate the role of murine cytomegalovirus GPCR M33 in the development of CAV in a murine aortic allograft model. MHC I-mismatched aortas of C.B10 (H2b) mice were transplanted into BALB/c (H2d) recipients, which were either mock-infected, infected with wild type (WT) MCMV or MCMV with a deleted M33-receptor gene (delM33). Persistence of cytomegalovirus infection was confirmed by qPCR and by luciferase assay to ensure active viral replication. Grafts were harvested on days 21 and 37 for intragraft mRNA expression and histological analysis. Active viral replication was demonstrated and MCMV presence was ensured by PCR within spleen, liver, salivary glands, lung and the aortic transplant. Infection with delM33 resulted in significantly less intimal proliferation compared to WT-MCMV but more pronounced proliferation than in mock-infected allografts (32.19% [delM33] vs. 41.71% [WT-MCMV] vs. 24.33% [MCMV-]). Intragraft expression of most analyzed genes was significantly increased in infected mice. VCAM-1, ICAM-1, PDGFβ, CXCR3 and Granzyme B were distinctly less expressed in grafts of delM33 infected compared to WT infected mice. Cellular infiltration revealed reduced dendritic cells and T cells in grafts infected with delM33 compared to WT MCMV. These data suggest that the MCMV encoded receptor M33 plays an important role as a viral effector mechanism contributing to the development of CAV in a murine aortic transplant model.
Niklas M. Fritz; Thomas Stamminger; Martina Ramsperger-Gleixner; Annika V. Kuckhahn; Regina Müller; Michael Weyand; Christian Heim. Cytomegalovirus chemokine receptor M33 knockout reduces chronic allograft rejection in a murine aortic transplant model. Transplant Immunology 2020, 64, 101359 .
AMA StyleNiklas M. Fritz, Thomas Stamminger, Martina Ramsperger-Gleixner, Annika V. Kuckhahn, Regina Müller, Michael Weyand, Christian Heim. Cytomegalovirus chemokine receptor M33 knockout reduces chronic allograft rejection in a murine aortic transplant model. Transplant Immunology. 2020; 64 ():101359.
Chicago/Turabian StyleNiklas M. Fritz; Thomas Stamminger; Martina Ramsperger-Gleixner; Annika V. Kuckhahn; Regina Müller; Michael Weyand; Christian Heim. 2020. "Cytomegalovirus chemokine receptor M33 knockout reduces chronic allograft rejection in a murine aortic transplant model." Transplant Immunology 64, no. : 101359.
Bernd Jahrsdörfer; Joris Kroschel; Carolin Ludwig; Victor Max Corman; Tatjana Schwarz; Sixten Körper; Markus Rojewski; Ramin Lotfi; Christof Weinstock; Christian Drosten; Erhard Seifried; Thomas Stamminger; Hans Jürgen Groß; Hubert Schrezenmeier. Independent side-by-side validation and comparison of four serological platforms for SARS-CoV-2 antibody testing. 2020, 1 .
AMA StyleBernd Jahrsdörfer, Joris Kroschel, Carolin Ludwig, Victor Max Corman, Tatjana Schwarz, Sixten Körper, Markus Rojewski, Ramin Lotfi, Christof Weinstock, Christian Drosten, Erhard Seifried, Thomas Stamminger, Hans Jürgen Groß, Hubert Schrezenmeier. Independent side-by-side validation and comparison of four serological platforms for SARS-CoV-2 antibody testing. . 2020; ():1.
Chicago/Turabian StyleBernd Jahrsdörfer; Joris Kroschel; Carolin Ludwig; Victor Max Corman; Tatjana Schwarz; Sixten Körper; Markus Rojewski; Ramin Lotfi; Christof Weinstock; Christian Drosten; Erhard Seifried; Thomas Stamminger; Hans Jürgen Groß; Hubert Schrezenmeier. 2020. "Independent side-by-side validation and comparison of four serological platforms for SARS-CoV-2 antibody testing." , no. : 1.
Pandemic SARS-CoV-2 infection has rapidly developed into a socioeconomic and humanitarian catastrophe. Basic principles to prevent SARS-CoV-2 transmission are social distancing, face masks, contact tracing and early detection of SARS-CoV-2. To meet these requirements, virtually unlimited test capacities delivering results in a rapid and reliable manner are a prerequisite. Here, we provide and validate such a rapid, convenient and efficient kit-independent detection of SARS-CoV-2 RNA, termed COVID-quick-DET. This straightforward method operates with simple proteinase K treatment and repetitive heating steps with a sensitivity of 94.6% in head-to-head comparisons with kit-based isolation methods. This result is supported by data obtained from serially diluted SARS-CoV-2 virus stocks. Given its cost- and time-effective operation, COVID-quick-DET might be best suited for countries with general shortage or temporary acute scarcity of resources and equipment.
Detlef Michel; Karin M. Danzer; Rüdiger Groß; Carina Conzelmann; Janis A. Müller; Axel Freischmidt; Jochen H. Weishaupt; Sandra Heller; Jan Münch; Manuela Michel; Thomas Stamminger; Alexander Kleger; Markus Otto. Rapid, convenient and efficient kit-independent detection of SARS-CoV-2 RNA. Journal of Virological Methods 2020, 286, 113965 -113965.
AMA StyleDetlef Michel, Karin M. Danzer, Rüdiger Groß, Carina Conzelmann, Janis A. Müller, Axel Freischmidt, Jochen H. Weishaupt, Sandra Heller, Jan Münch, Manuela Michel, Thomas Stamminger, Alexander Kleger, Markus Otto. Rapid, convenient and efficient kit-independent detection of SARS-CoV-2 RNA. Journal of Virological Methods. 2020; 286 ():113965-113965.
Chicago/Turabian StyleDetlef Michel; Karin M. Danzer; Rüdiger Groß; Carina Conzelmann; Janis A. Müller; Axel Freischmidt; Jochen H. Weishaupt; Sandra Heller; Jan Münch; Manuela Michel; Thomas Stamminger; Alexander Kleger; Markus Otto. 2020. "Rapid, convenient and efficient kit-independent detection of SARS-CoV-2 RNA." Journal of Virological Methods 286, no. : 113965-113965.
To avoid innate sensing and immune control, human immunodeficiency virus type 1 (HIV-1) has to prevent the accumulation of viral complementary DNA species. Here, we show that the late HIV-1 accessory protein Vpu hijacks DNA repair mechanisms to promote degradation of nuclear viral cDNA in cells that are already productively infected. Vpu achieves this by interacting with RanBP2–RanGAP1*SUMO1–Ubc9 SUMO E3-ligase complexes at the nuclear pore to reprogramme promyelocytic leukaemia protein nuclear bodies and reduce SUMOylation of Bloom syndrome protein, unleashing end degradation of viral cDNA. Concomitantly, Vpu inhibits RAD52-mediated homologous repair of viral cDNA, preventing the generation of dead-end circular forms of single copies of the long terminal repeat and permitting sustained nucleolytic attack. Our results identify Vpu as a key modulator of the DNA repair machinery. We show that Bloom syndrome protein eliminates nuclear HIV-1 cDNA and thereby suppresses immune sensing and proviral hyper-integration. Therapeutic targeting of DNA repair may facilitate the induction of antiviral immunity and suppress proviral integration replenishing latent HIV reservoirs.
Meta Volcic; Konstantin M. J. Sparrer; Lennart Koepke; Dominik Hotter; Daniel Sauter; Christina M. Stürzel; Myriam Scherer; Thomas Stamminger; Thomas G. Hofmann; Nathalie J. Arhel; Lisa Wiesmüller; Frank Kirchhoff. Vpu modulates DNA repair to suppress innate sensing and hyper-integration of HIV-1. Nature Microbiology 2020, 5, 1247 -1261.
AMA StyleMeta Volcic, Konstantin M. J. Sparrer, Lennart Koepke, Dominik Hotter, Daniel Sauter, Christina M. Stürzel, Myriam Scherer, Thomas Stamminger, Thomas G. Hofmann, Nathalie J. Arhel, Lisa Wiesmüller, Frank Kirchhoff. Vpu modulates DNA repair to suppress innate sensing and hyper-integration of HIV-1. Nature Microbiology. 2020; 5 (10):1247-1261.
Chicago/Turabian StyleMeta Volcic; Konstantin M. J. Sparrer; Lennart Koepke; Dominik Hotter; Daniel Sauter; Christina M. Stürzel; Myriam Scherer; Thomas Stamminger; Thomas G. Hofmann; Nathalie J. Arhel; Lisa Wiesmüller; Frank Kirchhoff. 2020. "Vpu modulates DNA repair to suppress innate sensing and hyper-integration of HIV-1." Nature Microbiology 5, no. 10: 1247-1261.
Nuclear egress is a rate-limiting step of herpesviral replication, restricting the nucleocytoplasmic transport of viral capsids. The process is regulated by two viral nuclear egress proteins (core NEC pUL50-pUL53), which recruit additional cellular and viral proteins. The multicomponent NEC mediates disassembly of the nuclear lamina barrier and the docking of nuclear capsids. The quantitation of nuclear egress has been accomplished by electron microscopic analysis, but is generally hampered by the low number of detectable cytoplasmic capsids. A newly established method for the quantitation of viral nuclear egress improves the characterization of viral mutants, host cell permissiveness and antiviral drug efficacy. In this study, various strains of human cytomegalovirus (HCMV) were used to measure the replication efficiencies in primary human fibroblasts, applying methods of cell fractionation, DNase digestion, sucrose cushions and quantitative PCR. Several stages of optimization led to a reliable quantitative assay that allowed the characterization of viral nuclear egress efficacy. Using this assay, recovery of the nuclear egress of a NEC-defective HCMV mutant was quantitatively assessed by applying an inducible NEC-expressing fibroblast culture for trans-complementation. This novel assay system can be further used to accurately quantitate and characterize the functionality of nuclear egress of HCMV or other herpesviruses.
Sigrun Häge; Deborah Horsch; Anne-Charlotte Stilp; Jintawee Kicuntod; Regina Müller; Stuart T. Hamilton; Ece Egilmezer; William D. Rawlinson; Thomas Stamminger; Eric Sonntag; Manfred Marschall. A quantitative nuclear egress assay to investigate the nucleocytoplasmic capsid release of human cytomegalovirus. Journal of Virological Methods 2020, 283, 113909 .
AMA StyleSigrun Häge, Deborah Horsch, Anne-Charlotte Stilp, Jintawee Kicuntod, Regina Müller, Stuart T. Hamilton, Ece Egilmezer, William D. Rawlinson, Thomas Stamminger, Eric Sonntag, Manfred Marschall. A quantitative nuclear egress assay to investigate the nucleocytoplasmic capsid release of human cytomegalovirus. Journal of Virological Methods. 2020; 283 ():113909.
Chicago/Turabian StyleSigrun Häge; Deborah Horsch; Anne-Charlotte Stilp; Jintawee Kicuntod; Regina Müller; Stuart T. Hamilton; Ece Egilmezer; William D. Rawlinson; Thomas Stamminger; Eric Sonntag; Manfred Marschall. 2020. "A quantitative nuclear egress assay to investigate the nucleocytoplasmic capsid release of human cytomegalovirus." Journal of Virological Methods 283, no. : 113909.
Human cytomegalovirus (HCMV) is the most frequent viral cause of congenital defects and can trigger devastating disease in immune-suppressed patients. Cytotoxic lymphocytes (CD8+ T cells and NK cells) control HCMV infection by releasing interferon-γ and five granzymes (GrA, GrB, GrH, GrK, GrM), which are believed to kill infected host cells through cleavage of intracellular death substrates. However, it has recently been demonstrated that the in vivo killing capacity of cytotoxic T cells is limited and multiple T cell hits are required to kill a single virus-infected cell. This raises the question whether cytotoxic lymphocytes can use granzymes to control HCMV infection in a noncytotoxic manner. Here, we demonstrate that (primary) cytotoxic lymphocytes can block HCMV dissemination independent of host cell death, and interferon-α/β/γ. Prior to killing, cytotoxic lymphocytes induce the degradation of viral immediate-early (IE) proteins IE1 and IE2 in HCMV-infected cells. Intriguingly, both IE1 and/or IE2 are directly proteolyzed by all human granzymes, with GrB and GrM being most efficient. GrB and GrM cleave IE1 after Asp398 and Leu414, respectively, likely resulting in IE1 aberrant cellular localization, IE1 instability, and functional impairment of IE1 to interfere with the JAK-STAT signaling pathway. Furthermore, GrB and GrM cleave IE2 after Asp184 and Leu173, respectively, resulting in IE2 aberrant cellular localization and functional abolishment of IE2 to transactivate the HCMV UL112 early promoter. Taken together, our data indicate that cytotoxic lymphocytes can also employ noncytotoxic ways to control HCMV infection, which may be explained by granzyme-mediated targeting of indispensable viral proteins during lytic infection. Human cytomegalovirus (HCMV) is the leading viral cause of congenital defects, can trigger disease in immune-compromised patients, and plays roles in cancer development. Cytotoxic lymphocytes kill HCMV-infected cells via releasing a set of five cytotoxic serine proteases called granzymes. However, the killing capacity of cytotoxic cells is limited and multiple T cell hits are required to kill a single virus-infected cell. This raises the question whether cytotoxic lymphocytes can use granzymes to control HCMV infection in a noncytotoxic manner. Here, we show that cytotoxic lymphocytes can also use granzymes to inhibit HCMV replication in absence of cell death. All five granzymes cleave and inactivate both viral immediate-early (IE1/2) proteins, which are essential players for initiating HCMV infection. Our data support the model that cytotoxic cells employ granzymes to dampen HCMV replication prior to accumulation of sufficient hits to kill the infected cell.
Liling Shan; Shuang Li; Jan Meeldijk; Bernadet Blijenberg; Astrid Hendriks; Karlijn J. W. M. Van Boxtel; Sara P. H. Van Den Berg; Ian J. Groves; Martin Potts; Adriana Svrlanska; Thomas Stamminger; Mark R. Wills; Niels Bovenschen. Killer cell proteases can target viral immediate-early proteins to control human cytomegalovirus infection in a noncytotoxic manner. PLOS Pathogens 2020, 16, e1008426 .
AMA StyleLiling Shan, Shuang Li, Jan Meeldijk, Bernadet Blijenberg, Astrid Hendriks, Karlijn J. W. M. Van Boxtel, Sara P. H. Van Den Berg, Ian J. Groves, Martin Potts, Adriana Svrlanska, Thomas Stamminger, Mark R. Wills, Niels Bovenschen. Killer cell proteases can target viral immediate-early proteins to control human cytomegalovirus infection in a noncytotoxic manner. PLOS Pathogens. 2020; 16 (4):e1008426.
Chicago/Turabian StyleLiling Shan; Shuang Li; Jan Meeldijk; Bernadet Blijenberg; Astrid Hendriks; Karlijn J. W. M. Van Boxtel; Sara P. H. Van Den Berg; Ian J. Groves; Martin Potts; Adriana Svrlanska; Thomas Stamminger; Mark R. Wills; Niels Bovenschen. 2020. "Killer cell proteases can target viral immediate-early proteins to control human cytomegalovirus infection in a noncytotoxic manner." PLOS Pathogens 16, no. 4: e1008426.
The multifunctional regulatory protein pUL69 of human cytomegalovirus acts as a viral RNA export factor with a critical role in efficient replication. Here, we identify serine/threonine phosphorylation sites for cellular and viral kinases within pUL69. We demonstrate that the pUL97/CDK phosphosites within alpha-helix 2 of pUL69 are crucial for its cis/trans isomerization by the cellular protein Pin1. Thus, we identified pUL69 as the first HCMV-encoded protein that is phosphorylated by cellular and viral serine/threonine kinases in order to serve as a substrate for Pin1. Furthermore, our study revealed that betaherpesviral mRNA export proteins contain extended binding motifs for the cellular mRNA adaptor proteins UAP56/URH49 harboring phosphorylated serines that are critical for efficient viral replication. Knowledge of the phosphorylation sites of pUL69 and the processes regulated by these posttranslational modifications is important in order to develop antiviral strategies based on a specific interference with pUL69 phosphorylation.
Marco Thomas; Regina Müller; Georg Horn; Boris Bogdanow; Koshi Imami; Jens Milbradt; Mirjam Steingruber; Manfred Marschall; Eva-Maria Schilling; Torgils Fossen; Thomas Stamminger. Phosphosite Analysis of the Cytomegaloviral mRNA Export Factor pUL69 Reveals Serines with Critical Importance for Recruitment of Cellular Proteins Pin1 and UAP56/URH49. Journal of Virology 2020, 94, 1 .
AMA StyleMarco Thomas, Regina Müller, Georg Horn, Boris Bogdanow, Koshi Imami, Jens Milbradt, Mirjam Steingruber, Manfred Marschall, Eva-Maria Schilling, Torgils Fossen, Thomas Stamminger. Phosphosite Analysis of the Cytomegaloviral mRNA Export Factor pUL69 Reveals Serines with Critical Importance for Recruitment of Cellular Proteins Pin1 and UAP56/URH49. Journal of Virology. 2020; 94 (8):1.
Chicago/Turabian StyleMarco Thomas; Regina Müller; Georg Horn; Boris Bogdanow; Koshi Imami; Jens Milbradt; Mirjam Steingruber; Manfred Marschall; Eva-Maria Schilling; Torgils Fossen; Thomas Stamminger. 2020. "Phosphosite Analysis of the Cytomegaloviral mRNA Export Factor pUL69 Reveals Serines with Critical Importance for Recruitment of Cellular Proteins Pin1 and UAP56/URH49." Journal of Virology 94, no. 8: 1.
Infections with human herpesviruses share several molecular characteristics, but the diversified medical outcomes are distinct to viral subfamilies and species. Notably, both clinical and molecular correlates of infection are a challenging field and distinct patterns of virus–host interaction have rarely been defined; this study therefore focuses on the search for virus-specific molecular indicators. As previous studies have demonstrated the impact of herpesvirus infections on changes in host signalling pathways, we illustrate virus-modulated expression levels of individual cellular protein kinases. Current data reveal (i) α-, β- and γ-herpesvirus-specific patterns of kinase modulation as well as (ii) differential levels of up-/downregulated kinase expression and phosphorylation, which collectively suggest (iii) defined signalling patterns specific for the various viruses (VSS) that may prove useful for defining molecular indicators. Combined, the study confirms the correlation between herpesviral replication and modulation of signalling kinases, possibly exploitable for the in vitro characterization of viral infections.
Manfred Marschall; Hanife Strojan; Richard Kiener; Christina Wangen; Eric Sonntag; Regina Müller; Isabel Zeitträger; Sabrina Wagner; Thomas Stamminger; Jens Milbradt; Uta Behrends; Nina Körber; Tanja Bauer; Silke Schrödel; Christian Thirion; Ralf Wagner; Corina Hutterer. Differential upregulation of host cell protein kinases by the replication of α-, β- and γ-herpesviruses provides a signature of virus-specific signalling. Journal of General Virology 2020, 101, 284 -289.
AMA StyleManfred Marschall, Hanife Strojan, Richard Kiener, Christina Wangen, Eric Sonntag, Regina Müller, Isabel Zeitträger, Sabrina Wagner, Thomas Stamminger, Jens Milbradt, Uta Behrends, Nina Körber, Tanja Bauer, Silke Schrödel, Christian Thirion, Ralf Wagner, Corina Hutterer. Differential upregulation of host cell protein kinases by the replication of α-, β- and γ-herpesviruses provides a signature of virus-specific signalling. Journal of General Virology. 2020; 101 (3):284-289.
Chicago/Turabian StyleManfred Marschall; Hanife Strojan; Richard Kiener; Christina Wangen; Eric Sonntag; Regina Müller; Isabel Zeitträger; Sabrina Wagner; Thomas Stamminger; Jens Milbradt; Uta Behrends; Nina Körber; Tanja Bauer; Silke Schrödel; Christian Thirion; Ralf Wagner; Corina Hutterer. 2020. "Differential upregulation of host cell protein kinases by the replication of α-, β- and γ-herpesviruses provides a signature of virus-specific signalling." Journal of General Virology 101, no. 3: 284-289.
The thrombopoietin receptor agonist eltrombopag was successfully used against human cytomegalovirus (HCMV)-associated thrombocytopenia refractory to immunomodulatory and antiviral drugs. These effects were ascribed to the effects of eltrombopag on megakaryocytes. Here, we tested whether eltrombopag may also exert direct antiviral effects. Therapeutic eltrombopag concentrations inhibited HCMV replication in human fibroblasts and adult mesenchymal stem cells infected with six different virus strains and drug-resistant clinical isolates. Eltrombopag also synergistically increased the anti-HCMV activity of the mainstay drug ganciclovir. Time-of-addition experiments suggested that eltrombopag interfered with HCMV replication after virus entry. Eltrombopag was effective in thrombopoietin receptor-negative cells, and the addition of Fe3+ prevented the anti-HCMV effects, indicating that it inhibits HCMV replication via iron chelation. This may be of particular interest for the treatment of cytopenias after hematopoietic stem cell transplantation, as HCMV reactivation is a major reason for transplantation failure. Since therapeutic eltrombopag concentrations are effective against drug-resistant viruses, and synergistically increase the effects of ganciclovir, eltrombopag is also a drug-repurposing candidate for the treatment of therapy-refractory HCMV disease.
Jens-Uwe Vogel; Sophie Schmidt; Daniel Schmidt; Florian Rothweiler; Benjamin Koch; Patrick Baer; Holger Rabenau; Detlef Michel; Thomas Stamminger; Martin Michaelis; Jindrich Cinatl; Schmidt. The Thrombopoietin Receptor Agonist Eltrombopag Inhibits Human Cytomegalovirus Replication Via Iron Chelation. Cells 2019, 9, 31 .
AMA StyleJens-Uwe Vogel, Sophie Schmidt, Daniel Schmidt, Florian Rothweiler, Benjamin Koch, Patrick Baer, Holger Rabenau, Detlef Michel, Thomas Stamminger, Martin Michaelis, Jindrich Cinatl, Schmidt. The Thrombopoietin Receptor Agonist Eltrombopag Inhibits Human Cytomegalovirus Replication Via Iron Chelation. Cells. 2019; 9 (1):31.
Chicago/Turabian StyleJens-Uwe Vogel; Sophie Schmidt; Daniel Schmidt; Florian Rothweiler; Benjamin Koch; Patrick Baer; Holger Rabenau; Detlef Michel; Thomas Stamminger; Martin Michaelis; Jindrich Cinatl; Schmidt. 2019. "The Thrombopoietin Receptor Agonist Eltrombopag Inhibits Human Cytomegalovirus Replication Via Iron Chelation." Cells 9, no. 1: 31.
Wedelolactone (WDL) is a coumestan present in the plants Eclipta prostrata and Wedelia calendulacea which are used for treatment of a multitude of health problems in traditional medicine. It has previously been shown that WDL exerts antiviral activity against human immunodeficiency virus and hepatitis C virus. In this study, we investigated the effect of WDL on lytic human cytomegalovirus (HCMV) infection. We demonstrate a strong interference with HCMV replication as analyzed in multi-round replication settings. A more detailed analysis of the underlying mechanisms revealed that WDL acts at two distinct steps of the viral replication cycle. During immediate early (IE) times, we observe an inhibition of IE1/IE2 expression. Although WDL was reported to interfere with NF-κB signaling our results suggest the existence of additional mechanisms that impede viral IE expression. During later time points of infection, WDL induced a disruption of the interaction between EZH2 and EED, components of the virus-supportive polycomb repressive complex 2 (PRC2). Thereby, the stability of the PRC2 complex as well as the related complex PRC1 was disturbed leading to diminished viral DNA synthesis. Taken together, we identify WDL as a potent agent against HCMV which interferes at two distinct steps of viral replication.
Adriana Svrlanska; Anna Ruhland; Manfred Marschall; Nina Reuter; Thomas Stamminger. Wedelolactone inhibits human cytomegalovirus replication by targeting distinct steps of the viral replication cycle. Antiviral Research 2019, 174, 104677 .
AMA StyleAdriana Svrlanska, Anna Ruhland, Manfred Marschall, Nina Reuter, Thomas Stamminger. Wedelolactone inhibits human cytomegalovirus replication by targeting distinct steps of the viral replication cycle. Antiviral Research. 2019; 174 ():104677.
Chicago/Turabian StyleAdriana Svrlanska; Anna Ruhland; Manfred Marschall; Nina Reuter; Thomas Stamminger. 2019. "Wedelolactone inhibits human cytomegalovirus replication by targeting distinct steps of the viral replication cycle." Antiviral Research 174, no. : 104677.