6:00 - 6:40
|
|
Plenary 3 - Viral Entry and Egress
Presenter:
|
Beate Sodeik, Doctor, Professor Institute of Virology, Hannover Medical School, Germany
Biography
Beate Sodeik obtained her Diploma in Biology with a thesis on autophagy in Amoeba proteus (1988, Bonn, Germany). As a DAAD Fellow, she then studied the interaction of human neutrophils with blood clots, and trained in immunology at Columbia University Medical School, New York, USA. In 1989, she moved to the European Molecular Biology Laboratory (EMBL, Heidelberg, Germany), and worked towards her PhD on the assembly of vaccinia virus. In 1993, she joined the group of Ari Helenius at Yale University (USA) as postdoctoral EMBO fellow and then staff scientist to establish novel cell biology and biochemistry approaches to investigate the early phase of herpes simplex virus infection. In 1997, she became lecturer at the Institute of Physiological Chemistry at Hannover Medical School, Germany, and established her own research team. After her "Habilitation" in Biochemistry, she was promoted to Assistant Professor in 2002 and to Full Professor in 2007 at the Institute of Virology at Hannover Medical School. Beate Sodeik holds also affiliations with the German Excellence Cluster RESIST and the German Center of Infection Research (DZIF) at Hannover Medical School, and currently serves her second term on the Study Section Virology of the German Research Foundation (DFG). She teaches virology, cell biology and biochemistry in the MSc and PhD programs of the Hannover Biomedical Research School.
|
A Hitchhikers Guide Through the Host Cell The World According to Capsids of Alphaherpesviruses
Beate Sodeik and her team study the cell biology of herpes simplex virus, a hitchhikers guide to epithelial cells, neurons, and immune cells. They investigate enabling virus-host cell interactions that that are required for efficient HSV-1 cell entry, nuclear targeting of incoming capsids and the release of viral genomes into the nucleoplasm as well as for the assembly of progeny capsids, nuclear capsid egress, and capsid envelopment. Furthermore, they characterize intrinsic and interferon-stimulated restricting host factors that perturb HSV-1 infection, and study HSV-1 proteins that counteract such host defense mechanisms, and have identified several small chemical compounds that interfere with capsid assembly, tegumentation and virion formation. The team uses BAC mutagenesis, state-of-the art biochemistry, life-cell imaging, conventional as well as quantitative immunoelectron microscopy, and mass spectrometry to elucidate the virus-host interactions during HSV-1 cell entry and assembly in keratinocytes, dendritic cells, macrophages, primary neurons ex vivo and in vivo infection of murine skin.
|
|
6:30 - 14:00
|
|
Virtual Help Desk
|
6:40 - 7:05
|
|
Symposium 3A - Viral Entry and Egress
Presenter:
|
Gill Elliott, Professor University of Surrey, United Kingdom
Biography
Gill Elliott is currently Professor of Virology at the University of Surrey, UK, where her group studies the cell biology of HSV1 infection. She obtained her PhD in the molecular biology of mumps virus from Queens University Belfast, and after a postdoctoral position studying molecular aspects of HIV transcription in Oxford, she began her career in herpesvirus research at the University of Leeds. A move to Marie Curie Research Institute, allowed her to pursue her interests in herpesvirus tegument proteins and assembly, initially in the group of Peter O'Hare, and subsequently in her own group where she pioneered the use of fluorescently-tagged viruses to study virus trafficking. In 2007, she moved to Imperial College London as an MRC Senior Fellow, and then to her current position at the University of Surrey in 2013. Her research has focused on virus morphogenesis and spread, with a particular interest in how HSV utilises the cellular secretory pathway to direct the process of assembly, with the aim of identifying cellular molecules crucial to this process. More recently, work on the tegument protein VP22 has also led to new understanding of spatial and temporal control of RNA metabolism during infection. She is keen to develop human keratinocytes as a relevant model for HSV1 infection, and having established a number of tools for this purpose her group is now investigating a range of virus-host interactions in these cells.
|
New Perspectives on the Nectin1-Glycoprotein D (gD) Complex from HSV1 Infected Human Keratinocytes
HSV1 infects the stratified epithelia of the epidermis, oral and genital mucosa, where the main cell type is the keratinocyte. In recent years, we have established human nTERT keratinocytes as a physiologically relevant model for studying HSV1 infection and have shown that HSV1 enters these cells within 5 minutes of attachment. This rapid entry correlates with high expression of the glycoprotein D (gD) receptor nectin1 on the cell surface of these cells, allowing the study of endogenous cell surface receptor by flow cytometry and immunofluorescence, rather than overexpressed nectin1. Using CRISPR-Cas9 knockout, we have confirmed that nectin1 is the major route of HSV1 and HSV2 entry into these cells, although an alternative, as yet unidentified route allows slower entry in to a small subset in the absence of nectin1. We have also found that during HSV1 or HSV2 infection, nectin1, but not nectin2, is re-organised, clustered and subsequently removed from the cell surface. Virus protein expression, and in particular, de novo expression of gD, is essential and sufficient for this nectin1 downregulation. Unusually for a membrane protein, this process involves proteosomal, not lysosomal degradation, but does not require the virus-encoded E3 ubiquitin ligase, ICP0. Consequently, we are currently screening cellular E3 ligases for their role in gD-induced nectin1 depletion during HSV1 infection. In this presentation I will further discuss the work we have been doing to investigate the purpose of HSV1-induced nectin1 downmodulation. Of note, work from ourselves and others has shown that additional viruses, including vaccinia virus and SARS-CoV2 also downregulate nectin1 during infection, suggesting that the studies presented here have implications for a wide range of viruses, and potentially highlight a novel, broadly applicable virus-host cell interaction.
|
|
7:05 - 7:30
|
|
Symposium 3B - Viral Entry and Egress
Presenter:
|
Jens von Einem, Professor Institute of Virology, Ulm University Hospital, Germany
Biography
Jens von Einem is a Professor at the Institute of Virology at the Ulm University Hospital. He obtained his PhD in Virology at the University of Greifswald for his research on herpesviruses at the Federal Research Institute for Animal Health. He continued his research on herpesvirus morphogenesis and pathogenesis as well as vaccine candidate development as a postdoctoral fellow at Cornell University with Klaus Osterrieder. In 2006, he took over the position of young group leader at the Institute of Virology in Ulm, where he was appointed Assistant Professor in 2011. His lab has been instrumental in using advanced electron microscopy to study herpesvirus morphogenesis, focusing on human cytomegalovirus. His group is investigating the molecular mechanisms underlying the assembly of infectious virus particles to identify critical steps that could serve as novel targets to limit herpesvirus dissemination.
|
Mechanistic Insights Into Secondary Envelopment
Virion morphogenesis is completed in the cytoplasm when tegumented capsids acquire their envelope by budding into cellular vesicles equipped with membrane embedded viral glycoproteins. This envelopment process, known as secondary envelopment, is crucial for the generation of infectious virions and very similar in all herpesviruses. Mechanistically, envelopment appears to be initiated, driven and completed by intertwined protein-protein and protein-membrane interactions. Although the general steps of human cytomegalovirus (HCMV) morphogenesis are well understood, the detailed molecular mechanisms are often not known. In our past and ongoing research, we have investigated several viral proteins, including tegument proteins (e.g. pUL71) and glycoproteins (e.g. gM and gN), and thoroughly characterized their role for HCMV secondary envelopment. These studies clearly support the idea of a sequential envelopment process that is regulated by different sets of viral proteins. The identification of several viral key players of secondary envelopment now provides the basis for studying regulatory functions and functional protein domains. This presentation will highlight some of our recent findings on the role of palmitoylation of viral proteins in the secondary envelopment process, indicating that palmitoylation is a central regulatory signal of different envelopment stages.
|
|
7:30 - 8:00
|
|
Networking Break/Meet the Speakers
|
8:00 - 9:30 - Concurrent Session
|
|
Concurrent Session 2A - Virus/Host Interactions 2
|
|
Concurrent Session 2B - Gene Expression and Replication 2
|
|
Concurrent Session 2C - Vaccines, Therapeutics and EPI
|
9:30 - 9:45
|
|
Networking Break
|
9:45 - 10:30
|
|
VZV Lecture
Presenter:
|
Trine Mogensen, Professor Department of Clinical Medicine Department of Infectious Diseases, Aarhus University, Denmark
Biography
Trine H. Mogensen obtained her Medical degree from Aarhus University (AU) in 2002, a PhD degree in 2003, and a Doctor of Medical Sciences degree (DMSc) in 2009. International training includes studies in Biochemistry and in Medicine in Paris, Descartes University and Necker Medical Faculty, Research fellow in the laboratory of Professor Bryan Williams at the Cleveland Clinic, Ohio, and a Diploma from the London School of Hygiene and Tropical Medicine. In 2014, she obtained her certificate as specialist doctor in infectious diseases. She became a professor in Infection Immunology at Department of Biomedicine, AU in 2017. Her current affiliations include Dept. of Biomedicine (AU) and Dept. of Infectious Diseases, Aarhus University Hospital, where she directs her research group and treats patients, respectively. Her clinical work and research focus on the genetic and immunological basis of primary immunodeficiencies and severe infectious diseases in humans, most notably viral infections in the central nervous system. Through a translational approach combining whole exome sequencing of patient samples with functional studies in molecular immunology and virology, her group aims at delineating the pathogenesis of infectious diseases, including herpes simplex encephalitis, recurrent HSV-2 meningitis, and VZV encephalitis with the ultimate goal to improve diagnosis, prevention, and treatment of these viral infections in the brain.
|
Inborn Errors of Immunity Predisposing to HSV Infections in the CNS
Herpesvirus infections can lead to a number of severe diseases, particularly when involving the central nervous system. However, understanding of the factors conferring susceptibility to these diseases and their complications remains incomplete. Previous studies have uncovered that defects in the innate Toll-like receptor 3 pathways in children and adults predispose to herpes simplex encephalitis by herpes simplex virus (HSV)1. Subsequently, our group described rare genetic variants/mutations in RNA polymerase III in children with varicella zoster virus (VZV) encephalitis demonstrating the importance of this innate cytosolic DNA sensor in antiviral defense against VZV.
In the present work we describe two adult patients with recurrent HSV2 lymphocytic (Mollarets) meningitis each carrying monoallelic rare potentially disease-causing variants in autophagy genes (ATG4A and LC3). Using a neuronal cell line we found that HSV2 was captured in autophagosomes, and blockage of autophagy by disruption of the ATG5 gene resulted in enhanced viral replication and cell death. Importantly, HSV2-activated autophagy was impaired in patient fibroblasts, which also exhibited significantly increased viral replication together with enhanced cell death. We found that HSV2-induced autophagy required viral late gene expression but was independent of the signaling adaptor molecule STING. Finally, reconstitution of wt allele expression of autophagy genes in cells from both patients restored virus-activated autophagy and the ability to control HSV2 replication. Altogether, we suggest an important role for autophagy in anti-HSV2 immunity in the nervous system. This represents the first description of an autophagy defect causing a primary immunodeficiency conferring increased susceptibility to viral CNS infection in humans.
|
|
10:30 - 11:10
|
|
Plenary 4 - Gene Expression and Epigenetics
Presenter:
|
Neal DeLuca, Professor University of Pittsburgh School of Medicine, United States
Biography
Neal DeLuca did his undergraduate studies at Lehigh University where he received a B.S. in Fundamental Sciences in 1977. He performed his graduate studies at The Pennsylvania State University and received M.S. and PhD degrees in Biophysics in 1979 and 1981, respectively. Dr. DeLuca went on to do postdoctoral research with Dr. Pricilla Schaffer at the Dana Farber Cancer Institute and Harvard Medical School from 1982-1986. He stayed on as an Assistant Professor in Department of Microbiology and Molecular Genetics Harvard Medical School, at the Dana Farber Cancer Institute from 1986-1991. In 1991 Dr. DeLuca moved to the University of Pittsburgh School of Medicine where he is currently a Professor of Microbiology and Molecular Genetics. Dr. DeLuca has served as a member of the Virology and Virology A Study Sections, The NIH Recombinant DNA Advisory Committee, and the NIAID MID Study Section, as Chair. He is a Fellow of the American Academy of Microbiology.
|
HSV-1 ICP4 as Viral Nucleoprotein and Transcription Factor
The gene encoding ICP4 is one of five HSV immediate early genes that are transcribed in the absence of prior viral protein synthesis by virtue of the virion transcription factor VP16. ICP4 is required for progression to the next phases of viral transcription. Early in infection, ICP4 densely binds to the viral genome, forming a type of nucleoprotein. This nucleoprotein is distinct from the chromatin structure that one observes for the cellular genome. As viral DNA replication proceeds more ICP4 binds to the genome, albeit less densely. How these changes affect the transcription of the genome is not known, however the result is the activation of late transcription and the shutoff of most cellular transcription. In the absence of ICP4, early and late transcription is greatly attenuated and genome replication doesnt occur. Viral mutants that lack functional ICP4 can be propagated in cells that supply ICP4 in trans allowing for the study of the requirement for ICP4. However, these virus contain wt ICP4 in their tegument, and the fate of this source of ICP4 is unknown. Another fate of ICP4 expressed in infected cells is the binding to euchromatic sites in the cellular genome. ICP4 does not bind to heterochromatic sites, and as viral DNA replication progresses the binding of ICP4 to the cellular genome is reduced. If and how ICP4 contributes to the differential cellular gene expression seen early in infection is not known.
|
|
11:10 - 11:35
|
|
Symposium 4A - Gene Expression and Epigenetics
Presenter:
|
Matthew Reeves, Doctor University College London, United Kingdom
Biography
Matthew Reeves currently leads a research laboratory based at the Institute of Immunity & Transplantation, UCL, UK where his lab investigates the molecular details of the host:pathogen interaction with a view to understanding pathogenesis in vivo and developing strategies to prevent it.
His longstanding interest in HCMV concerns the biology of latency and, specifically, the importance of chromatin and cell signalling in dendritic cells as a driver of reactivation. Additionally, Matthew has investigated the role of cell death in both lytic and latent infection and developed an interest in gB through these studies. Recent work in the lab is centred on trying to understand the humoral immune response to the gB vaccine alongside trying to decipher key components of tissue resident immunity in transplanted organs.
|
Inflammatory Signaling, Chromatin and Human Cytomegalovirus Reactivation
Human cytomegalovirus (HCMV) latency and reactivation is regulated by a complex interplay of host cell signaling, epigenetics and cell identity. An important site of HCMV latency is the CD34+ haematopoietic progenitor cell which, upon differentiation to a myeloid dendritic cell (DC) or macrophage, can support viral reactivation. Silencing of the major immediate early (MIE) locus has long been considered crucial for the establishment of latency and, by extension, the reversal of this silencing required for reactivation.
In this overview, I will discuss the contributions of many research groups that have led to the current model of HCMV reactivation and then focus on our own studies seeking to understand how inflammatory signals promote HCMV reactivation in DCs through a complex interplay of multiple signal pathways. A particular emphasis will be on the key role dynamic regulation of chromatin plays in this process and thus consider why cell identity is important for mediating these changes with a view to novel therapeutic approaches to abrogate the reactivation of this important medical pathogen.
|
|
11:35 - 12:00
|
|
Symposium 4B - Gene Expression and Epigenetics
Presenter:
|
Italo Tempera, Associate Professor The Wistar Institute, United States
Biography
Italo Tempera is an Associate Professor at The Wistar Institute in Philadelphia. He obtained his Ph.D. degree in Biochemistry from the University of Rome La Sapienza where he studied the role of histone modifications in EBV infection. Upon completing his Ph.D., he continued his training in the laboratory of Dr. Paul Lieberman at The Wistar Institute, where he studied how the three-dimensional structure of the viral genome controls EBV gene expression during latency. In 2012, he started his independent laboratory at the Fels Institute for Cancer Research at Temple University. In 2020, he moved to the Wistar Institute, where his lab continues to study how EBV and host each attempt to alter the 3-dimensional structure and the composition of the others chromatin to achieve their own ends.
|
3D Genome Remodeling During EBV Latency
In latent infection, EBV expresses distinct subsets of viral genes in various patterns, referred to as latency types. These different latencies reflect gene programs that transition during EBV infection and the establishment of latency. Epigenetic modifications play an essential role in regulating EBV gene expression during latency, and the pattern of epigenetic markers that decorate the viral episome differs between latency types. Studies from others and our group demonstrated that these epigenetic patterns are maintained across the EBV genome by the cellular factor CTCF. CTCF is a highly conserved DNA binding protein that regulates several processes, including chromatin insulation and looping. We have previously shown that viral latent promoters can form chromatin loops through CTCF in B cells, suggesting that chromatin looping is an essential regulator of EBV latency. To better understand the role of 3D genome remodeling in regulating EBV latency, we have developed an EBV capture-Hi-C assay that allows us to generate 3D maps of the EBV genome at 5kb resolution. By systematically applying our methods to the viral episome in epithelial and infected B cells, we generated the first complete 3D map of the EBV genome in different latency programs. We find that the 3D genome organization of EBV is dynamic and correlates with the viral transcription and origin of infected cells. We observe that the maintenance and folding of these viral 3D chromatin structures requires the binding of CTCF which is regulated through PARP1. Changes in PARP1 activity or metabolic pathways linked to PARP1 functions alter the EBV 3D genome. By extending our approach to the host, we mechanistically observe similar dynamic fluctuations of the 3D genome due to CTCF and PARP1 interactions. Overall, these data illustrate the essential role of 3D genome remodeling in regulating EBV latency and provide novel insights into cellular mechanisms that epigenetically regulate EBV infection.
|
|
12:00 - 12:30
|
|
Meet the Speakers
|
12:00 - 14:00
|
|
Poster Session 1
|