IM-4 Impact of oHSV activated NOTCH signaling in tumor microenvironment and its impact on anti-tumor immunity

Abstract Oncolytic herpes simplex virus-1 (oHSV) is novel FDA-approved immunotherapy for advanced melanoma patients in US. Also, oHSV is recently approved for the treatment of recurrent GBM in Japan. We have shown that oHSV treatment of GBM cells induces NICD cleavage and NOTCH activation in adjacent uninfected glioma cells via HSV-1 microRNA-H16 (Otani Y and Yoo JY, Clin Cancer Res, 2020), however, the consequences of NOTCH on immunotherapy in GBM is unknow. Here we have investigated the impact of oHSV-induced NOTCH signaling on the tumor microenvironment (TME). Analysis of TCGA GBM data and experimental murine models revealed NOTCH induced immunosuppressive myeloid cell recruitment and limited anti-tumor immunity. In oHSV treated tissue, viral infection educated tumor associated macrophages to secrete CCL2 which recruited monocytic myeloid derived suppressor cell (MDSC) that attenuated anti-tumor immunity. Consistent with this, CCL2 induction was also observed in serum of recurrent GBM patients treated with oHSV (NCT03152318). Importantly, blockade of NOTCH signaling reduced the oHSV induced immunosuppressive environment and activated a CD8 dependent anti-tumor memory response. These findings present the opportunities for combination therapies that can help improve therapeutic benefit and anti-tumor immunity in GBM.

Download Full-text

TAMI-11. IMPACT OF oHSV ACTIVATED NOTCH SIGNALING IN TUMOR MICROENVIRONMENT, AND ITS IMPACT ON ANTI-TUMOR IMMUNITY

Neuro-Oncology ◽

10.1093/neuonc/noaa215.900 ◽

2020 ◽

Vol 22 (Supplement_2) ◽

pp. ii215-ii215

Author(s):

Yoshihiro Otani ◽

Ji Young Yoo ◽

Samantha Chao ◽

Toshihiko Shimizu ◽

Cole Lewis ◽

...

Keyword(s):

Tumor Microenvironment ◽

Notch Signaling ◽

Tumor Immunity ◽

Cell Communication ◽

Notch Signaling Pathway ◽

Therapeutic Benefit ◽

Notch Receptor ◽

Negative Regulator ◽

Gamma Secretase ◽

The Impact

Abstract NOTCH signaling is a method of cell-cell communication where membrane bound NOTCH ligands on signal-sending cells can bind to and initiate cleavage of the NOTCH receptor, releasing NICD which can initiate signal transduction in adjacent “signal-receiving” cells. We have recently shown that oHSV treatment of GBM cells induces NICD cleavage and NOTCH activation in adjacent uninfected glioma cells. RNA sequencing of GBM cells post-infection also uncovered Gene Ontology NOTCH signaling pathway to be significantly upregulated. This activation was induced by viral miRNA-H16, which represses FIH-1 expression. FIH-1 was found to be a negative regulator of Mib1, a ubiquitin ligase, which activates NOTCH ligand-mediated activation of adjacent signal-receiving cells bearing the NOTCH receptor (Otani et al Clin. Can. Res. 2020). Here we have investigated the impact of oHSV-induced NOTCH signaling on the tumor microenvironment. Treatment of brain tumors in immune competent mice with oHSV and NOTCH blocking gamma secretase inhibitor (GSI) induced an anti-tumor memory immune response. Long term survivors in mice treated with the combination also completely rejected subsequent tumor re-challenge in the other hemisphere. UMAP of flow cytometry of tumor-bearing hemispheres and functional analysis of isolated cellular fractions from treated mice showed a significant influx of MDSC cells after oHSV treatment that was rescued in mice treated with oHSV and GSI. Ongoing mechanistic studies are uncovering a significant induction of NOTCH in tumor associated macrophages that aids in recruitment of MDSC cells. Overall these studies have uncovered a significant impact of oHSV therapy on GBM tumor microenvironment and presents opportunities for combination therapies that can help improve therapeutic benefit and anti-tumor immunity.

Download Full-text

695 Oral delivery of a microbial extracellular vesicle induces potent anti-tumor immunity in mice

Journal for ImmunoTherapy of Cancer ◽

10.1136/jitc-2020-sitc2020.0695 ◽

2020 ◽

Vol 8 (Suppl 3) ◽

pp. A737-A737

Author(s):

Loise Francisco-Anderson ◽

Mary Abdou ◽

Michael Goldberg ◽

Erin Troy ◽

...

Keyword(s):

Tumor Microenvironment ◽

Tumor Growth ◽

Extracellular Vesicles ◽

Tumor Immunity ◽

Extracellular Vesicle ◽

The Body ◽

Checkpoint Inhibition ◽

Small Intestinal ◽

The Impact

BackgroundThe small intestinal axis (SINTAX) is a network of anatomic and functional connections between the small intestine and the rest of the body. It acts as an immunosurveillance system, integrating signals from the environment that affect physiological processes throughout the body. The impact of events in the gut in the control of tumor immunity is beginning to be appreciated. We have previously shown that an orally delivered single strain of commensal bacteria induces anti-tumor immunity preclinically via pattern recognition receptor-mediated activation of innate and adaptive immunity. Some bacteria produce extracellular vesicles (EVs) that share molecular content with the parent bacterium in a particle that is roughly 1/1000th the volume in a non-replicating form. We report here an orally-delivered and gut-restricted bacterial EV which potently attenuates tumor growth to a greater extent than whole bacteria or checkpoint inhibition.MethodsEDP1908 is a preparation of extracellular vesicles produced by a gram-stain negative strain of bacterium of the Oscillospiraceae family isolated from a human donor. EDP1908 was selected for its immunostimulatory profile in a screen of EVs from a range of distinct microbial strains. Its mechanism of action was determined by ex vivo analysis of the tumor microenvironment (TME) and by in vitro functional studies with murine and human cells.ResultsOral treatment of tumor-bearing mice with EDP1908 shows superior control of tumor growth compared to checkpoint inhibition (anti-PD-1) or an intact microbe. EDP1908 significantly increased the percentage of IFNγ and TNF producing CD8+ CTLs, NK cells, NKT cells and CD4+ cells in the tumor microenvironment (TME). EDP1908 also increased tumor-infiltrating dendritic cells (DC1 and DC2). Analysis of cytokines in the TME showed significant increases in IP-10 and IFNg production in mice treated with EDP1908, creating an environment conducive to the recruitment and activation of anti-tumor lymphocytes.ConclusionsThis is the first report of striking anti-tumor effects of an orally delivered microbial extracellular vesicle. These data point to oral EVs as a new class of immunotherapeutic drugs. They are particularly effective at harnessing the biology of the small intestinal axis, acting locally on host cells in the gut to control distal immune responses within the TME. EDP1908 is in preclinical development for the treatment of cancer.Ethics ApprovalPreclinical murine studies were conducted under the approval of the Avastus Preclinical Services’ Ethics Board. Human in vitro samples were attained by approval of the IntegReview Ethics Board; informed consent was obtained from all subjects.

Download Full-text

Distinct Tumor Microenvironments Are a Defining Feature of Strain-Specific CRISPR/Cas9-Induced MPNSTs

Genes ◽

10.3390/genes11050583 ◽

2020 ◽

Vol 11 (5) ◽

pp. 583 ◽

Cited By ~ 2

Author(s):

Amanda Scherer ◽

Victoria R. Stephens ◽

Gavin R. McGivney ◽

Wade R. Gutierrez ◽

Emily A. Laverty ◽

...

Keyword(s):

Tumor Microenvironment ◽

Mouse Models ◽

Cancer Biology ◽

Myeloid Cell ◽

Inbred Strains ◽

Peripheral Nerve Sheath Tumors ◽

Tumor Onset ◽

The Impact ◽

Strain Dependent

The tumor microenvironment plays important roles in cancer biology, but genetic backgrounds of mouse models can complicate interpretation of tumor phenotypes. A deeper understanding of strain-dependent influences on the tumor microenvironment of genetically-identical tumors is critical to exploring genotype–phenotype relationships, but these interactions can be difficult to identify using traditional Cre/loxP approaches. Here, we use somatic CRISPR/Cas9 tumorigenesis approaches to determine the impact of mouse background on the biology of genetically-identical malignant peripheral nerve sheath tumors (MPNSTs) in four commonly-used inbred strains. To our knowledge, this is the first study to systematically evaluate the impact of host strain on CRISPR/Cas9-generated mouse models. Our data identify multiple strain-dependent phenotypes, including changes in tumor onset and the immune microenvironment. While BALB/c mice develop MPNSTs earlier than other strains, similar tumor onset is observed in C57BL/6, 129X1 and 129/SvJae mice. Indel pattern analysis demonstrates that indel frequency, type and size are similar across all genetic backgrounds. Gene expression and IHC analysis identify multiple strain-dependent differences in CD4+ T cell infiltration and myeloid cell populations, including M2 macrophages and mast cells. These data highlight important strain-specific phenotypes of genomically-matched MPNSTs that have implications for the design of future studies using similar in vivo gene editing approaches.

Download Full-text

Efficacy and Tolerance of Thymus Vulgaris Extract in Patients with Covid-19

Journal of Physical Medicine Rehabilitation Studies & Reports ◽

10.47363/jpmrs/2021(3)145 ◽

2021 ◽

pp. 1-5

Author(s):

Yiagnigni Mfopou E ◽

Keyword(s):

Hot Water ◽

Thymus Vulgaris ◽

Herpes Simplex Virus 1 ◽

Virus Elimination ◽

Good Tolerance ◽

Negative Results ◽

Simplex Virus ◽

The Impact ◽

Blood Urea Level ◽

Positive Effect

The coronavirus disease 19 (COVID-19) has been a pandemic since February 2020. So far, no effective treatment has been found. WHO has recommended research on medicinal plants as an alternative treatment course. Several studies conducted on Thymus vulgaris have established its antioxidant, antiviral and immunomodulatory properties that induce the elimination of viruses such as Herpes simplex Virus 1 and 2. Following this, we initiated a study entitled Efficacy and tolerance of Thymus vulgaris extract in patients with coronavirus 2019. Material and method: Thymus vulgaris powder was used in this study. A consent letter and a questionnaire about the patients’ symptoms were prepared to be used by a research investigator. According to the statistical calculations of this cohort study, 161 patients testing positive for COVID-19 PCR were consecutively recruited, of which 75 patients were not exposed to Thymus vulgaris and 86 patients were exposed. Information from the questionnaire was gathered from the patients before the initiation of conventional treatment (vitamin C 1000 1 tablet/day, Zinc 20 mg 1tablet/day, Azithromycin 500 1cp day and amoxicillin/clavulamic acid 1g/125 1 tablet per 12 hours for six days in both cohorts) and by combining Thymus vulgaris (1 teaspoon, i.e. 5g, in 100 ml of hot water to be taken every 8 hours) by the patients in the exposed cohort. After three days of this treatment, the evaluative part of the questionnaire was completed to assess the impact of taking or not taking Thymus vulgaris on early symptoms and tolerance; on the 10th day after the start of treatment, the PCR control test was carried out. Thereafter, the various statistical analyses were performed. Results: Statistical evaluation after three days of treatment shows that taking Thymus vulgaris has a statistically significant positive effect on cough (p <0.01), dyspnoea (p <0.001), dizziness (p <0.029), fatigue (p <0.001), anorexia (p <0.001), chest pain (p <0.001), fever (p <0.024), agueusia (p <0.029) and anosmia (p <0.001). There was a significant decrease in neutrophils (p <0.01); in addition, the lymphocyte count increased significantly (p <0.001) as did the serum calcium level (p <0.03). Blood urea level decreased significantly (p <0.01). Significant negative results of the COVID-19 PCR were obtained at Day 10 in the exposed group (p <0.001). In addition, there was no significant change in other biological parameters such as creatinine, blood glucose, aspartate amino transferase. Conclusion: Results of this study show that the use of the powder of Thymus vulgaris, a medicinal plant, with antioxidant, immunomodulatory and antiviral properties, was very effective on coronavirus-induced symptoms and virus elimination. Moreover, there was good tolerance after taking Thymus vulgaris.

Download Full-text

Transcriptome Analysis of the Cerebellum of Mice Fed a Manganese-Deficient Diet

Frontiers in Genetics ◽

10.3389/fgene.2020.558725 ◽

2020 ◽

Vol 11 ◽

Author(s):

Young Ah Seo ◽

Eun-Kyung Choi ◽

Luisa Aring ◽

Molly Paschall ◽

Shigeki Iwase

Keyword(s):

Gene Expression ◽

Cerebellar Atrophy ◽

Epidemiological Studies ◽

Receptor Interaction ◽

Herpes Simplex Virus 1 ◽

Deficient Diet ◽

Mn Deficiency ◽

Metal Levels ◽

Simplex Virus ◽

The Impact

Manganese (Mn), primarily acquired through diet, is required for brain function and development. Epidemiological studies have found an association between both low and high levels of Mn and impaired neurodevelopment in children. Recent genetic studies have revealed that patients with congenital Mn deficiency display severe psychomotor disability and cerebral and cerebellar atrophy. Although the impact of Mn on gene expression is beginning to be appreciated, Mn-dependent gene expression remains to be explored in vertebrate animals. The goal of this study was to use a mouse model to define the impact of a low-Mn diet on brain metal levels and gene expression. We interrogated gene expression changes in the Mn-deficient mouse brain at the genome-wide scale by RNA-seq analysis of the cerebellum of mice fed low or normal Mn diets. A total of 137 genes were differentially expressed in Mn-deficient cerebellums compared with Mn-adequate cerebellums (Padj < 0.05). Mn-deficient mice displayed downregulation of key pathways involved with “focal adhesion,” “neuroactive ligand-receptor interaction,” and “cytokine-cytokine receptor interaction” and upregulation of “herpes simplex virus 1 infection,” “spliceosome,” and “FoxO signaling pathway.” Reactome pathway analysis identified upregulation of the splicing-related pathways and transcription-related pathways, as well as downregulation of “metabolism of carbohydrate,” and “extracellular matrix organization,” and “fatty acid metabolism” reactomes. The recurrent identifications of splicing-related pathways suggest that Mn deficiency leads to upregulation of splicing machineries and downregulation of diverse biological pathways.

Download Full-text

Endocytic Internalization of Herpes Simplex Virus 1 in Human Keratinocytes at Low Temperature

Journal of Virology ◽

10.1128/jvi.02195-20 ◽

2020 ◽

Author(s):

Nydia De La Cruz ◽

Dagmar Knebel-Mörsdorf

Keyword(s):

Plasma Membrane ◽

Herpes Simplex Virus ◽

Herpes Simplex ◽

Low Temperature ◽

Human Keratinocytes ◽

Herpes Simplex Virus 1 ◽

Successful Infection ◽

Simplex Virus ◽

The Impact ◽

Hsv 1

Herpes simplex virus 1 (HSV-1) can adopt a variety of pathways to accomplish cellular internalization. In human keratinocytes representing the natural target cell of HSV-1, both direct plasma membrane fusion and endocytic uptake have been found. The impact of either pathway in successful infection, however, remains to be fully understood. To address the role of each internalization mode, we performed infection studies at low temperature as a tool to interfere with endocytic pathways. Interestingly, successful HSV-1 entry in primary human keratinocytes and HaCaT cells was observed even at 7°C, although delayed compared to infection at 37°C. Moreover, ex vivo infection of murine epidermis demonstrated that virus entry at 7°C is not only accomplished in cultured cells but also in tissue. Control experiments with cholera toxin B confirmed a block of endocytic uptake at 7°C. In addition, uptake of dextran by macropinosomes and phagocytic uptake of latex beads was also inhibited at 7°C. Infection of nectin-1-deficient murine keratinocytes affirmed that the entry at 7°C was receptor-dependent. Strikingly, the lysosomotropic agent, ammonium chloride, strongly inhibited HSV-1 entry suggesting a role for endosomal acidification. Ultrastructural analyses in turn revealed free capsids in the cytoplasm as well as virus particles in vesicles after infection at 7°C supporting both plasma membrane fusion and endocytic internalization as already observed at 37°C. Overall, entry of HSV-1 at 7°C suggests that the virus can efficiently adopt nectin-1-dependent unconventional vesicle uptake mechanisms in keratinocytes strengthening the role of endocytic internalization for successful infection. IMPORTANCE The human pathogen herpes simplex virus 1 (HSV-1) relies on multiple internalization pathways to initiate infection. Our focus is on the entry in human keratinocytes, the major in vivo target during primary and recurrent infection. While antivirals reduce the severity of clinical cases, there is no cure or vaccine against HSV. To develop strategies that interfere with virus penetration, we need to understand the various parameters and conditions that determine virus entry. Here, we addressed the impact of virus internalization via vesicles by blocking endocytic processes at low temperature. Intriguingly, we detected entry of HSV-1 even at 7°C which led to infection of primary keratinocytes and epidermal tissue. Moreover, electron microscopy of human keratinocytes at 7°C support that internalization is based on fusion of the viral envelope with the plasma membrane as well as vesicle membranes. These results provide novel insights into conditions that still allow endocytic internalization of HSV-1.

Download Full-text

Impact of Amino Acid Substitutions in Region II and Helix K of HSV-1 and HCMV DNA Polymerases on Resistance to Foscarnet

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00390-21 ◽

2021 ◽

Author(s):

Karima Zarrouk ◽

Xiaojun Zhu ◽

Van Dung Pham ◽

Nathalie Goyette ◽

Jocelyne Piret ◽

...

Keyword(s):

Amino Acid ◽

Conformational Changes ◽

Recombinant Dna ◽

Three Dimensional ◽

Amino Acid Substitutions ◽

Herpes Simplex Virus 1 ◽

Simplex Virus ◽

The Impact ◽

Region Ii ◽

Hsv 1

Amino acid substitutions conferring resistance of herpes simplex virus 1 (HSV-1) and human cytomegalovirus (HCMV) to foscarnet (PFA) are respectively located in UL30 and UL54 genes encoding the DNA polymerase (pol). In this study, we analyzed the impact of substitutions located in helix K and region II that are involved in the conformational changes of the DNA pol. Theoretical substitutions were identified by sequences alignment of the helix K and region II of human herpesviruses (susceptible to PFA) and bacteriophages (resistant to PFA) and introduced in viral genomes by recombinant phenotyping. We characterized the susceptibility of HSV-1 and HCMV mutants to PFA. In UL30, substitutions I619K (helix K), V715S and A719T (both in region II) increased mean PFA EC50 by 2.5-, 5.6- and 2.0-fold compared to wild type (WT), respectively. In UL54, substitution Q579I (helix K) conferred hypersusceptibility to PFA (0.17-fold change) whereas substitutions Q697P, V715S and A719T (all in region II) increased mean PFA EC50 values by 3.8-, 2.8- and 2.5-fold compared to WT, respectively. These results were confirmed by enzymatic assays using recombinant DNA pol harboring these substitutions. Three-dimensional modeling suggests that substitutions conferring resistance/hypersusceptibility to PFA located in helix K and region II of UL30 and UL54 DNA pol favor an open/closed conformation of these enzymes resulting in a lower/higher drug affinity for the proteins. Thus, this study shows that both regions of UL30 and UL54 DNA pol are involved in the conformational changes of these proteins and can influence the susceptibility of both viruses to PFA.

Download Full-text

The Impact of Radiation on the Tumor Microenvironment: Effect of Dose and Fractionation Schedules

Cancer Growth and Metastasis ◽

10.1177/1179064418761639 ◽

2018 ◽

Vol 11 ◽

pp. 117906441876163 ◽

Cited By ~ 39

Author(s):

Kimberly M Arnold ◽

Nicole J Flynn ◽

Adam Raben ◽

Lindsay Romak ◽

Yan Yu ◽

...

Keyword(s):

Tumor Microenvironment ◽

External Beam Radiotherapy ◽

Tumor Stroma ◽

Therapeutic Benefit ◽

Normal Tissues ◽

Technological Advances ◽

Fractionation Schedule ◽

Effects Of Radiation ◽

Hypofractionated Radiation ◽

The Impact

In addition to inducing lethal DNA damage in tumor and stromal cells, radiation can alter the interactions of tumor cells with their microenvironment. Recent technological advances in planning and delivery of external beam radiotherapy have allowed delivery of larger doses per fraction (hypofractionation) while minimizing dose to normal tissues with higher precision. The effects of radiation on the tumor microenvironment vary with dose and fractionation schedule. In this review, we summarize the effects of conventional and hypofractionated radiation regimens on the immune system and tumor stroma. We discuss how these interactions may provide therapeutic benefit in combination with targeted therapies. Understanding the differential effects of radiation dose and fractionation can have implications for choice of combination therapies.

Download Full-text

Entry Mechanisms of Herpes Simplex Virus 1 into Murine Epidermis: Involvement of Nectin-1 and Herpesvirus Entry Mediator as Cellular Receptors

Journal of Virology ◽

10.1128/jvi.02917-14 ◽

2014 ◽

Vol 89 (1) ◽

pp. 262-274 ◽

Cited By ~ 23

Author(s):

Philipp Petermann ◽

Katharina Thier ◽

Elena Rahn ◽

Frazer J. Rixon ◽

Wilhelm Bloch ◽

...

Keyword(s):

Herpes Simplex Virus ◽

Herpes Simplex ◽

Ex Vivo ◽

Target Tissue ◽

Herpes Simplex Virus 1 ◽

Simplex Virus ◽

The Impact ◽

Herpesvirus Entry Mediator ◽

Hsv 1

ABSTRACTSkin keratinocytes represent a primary entry site for herpes simplex virus 1 (HSV-1)in vivo. The cellular proteins nectin-1 and herpesvirus entry mediator (HVEM) act as efficient receptors for both serotypes of HSV and are sufficient for disease development mediated by HSV-2 in mice. How HSV-1 enters skin and whether both nectin-1 and HVEM are involved are not known. We addressed the impact of nectin-1 during entry of HSV-1 into murine epidermis and investigated the putative contribution of HVEM. Usingex vivoinfection of murine epidermis, we showed that HSV-1 entered the basal keratinocytes of the epidermis very efficiently. In nectin-1-deficient epidermis, entry was strongly reduced. Almost no entry was observed, however, in nectin-1-deficient keratinocytes grown in culture. This observation correlated with the presence of HVEM on the keratinocyte surface in epidermis and with the lack of HVEM expression in nectin-1-deficient primary keratinocytes. Our results suggest that nectin-1 is the primary receptor in epidermis, while HVEM has a more limited role. For primary murine keratinocytes, on which nectin-1 acts as a single receptor, electron microscopy suggested that HSV-1 can enter both by direct fusion with the plasma membrane and via endocytic vesicles. Thus, we concluded that nectin-1 directs internalization into keratinocytes via alternative pathways. In summary, HSV-1 entry into epidermis was shown to strongly depend on the presence of nectin-1, but the restricted presence of HVEM can potentially replace nectin-1 as a receptor, illustrating the flexibility employed by HSV-1 to efficiently invade tissuein vivo.IMPORTANCEHerpes simplex virus (HSV) can cause a range of diseases in humans, from uncomplicated mucocutaneous lesions to life-threatening infections. The skin is one target tissue of HSV, and the question of how the virus overcomes the protective skin barrier and penetrates into the tissue to reach its receptors is still open. Previous studies analyzing entry into cells grownin vitrorevealed nectin-1 and HVEM as HSV receptors. To explore the contributions of nectin-1 and HVEM to entry into a natural target tissue, we established anex vivoinfection model. Using nectin-1- or HVEM-deficient mice, we demonstrated the distinct involvement of nectin-1 and HVEM for HSV-1 entry into epidermis and characterized the internalization pathways. Such advances in understanding the involvement of receptors in tissue are essential preconditions for unraveling HSV invasion of skin, which in turn will allow the development of antiviral reagents.

Download Full-text

Spatiotemporal analysis of host cell modification during herpes simplex virus 1 replication

10.1101/2019.12.22.872002 ◽

2019 ◽

Cited By ~ 1

Author(s):

Katharina M. Scherer ◽

James D. Manton ◽

Timothy K. Soh ◽

Luca Mascheroni ◽

Viv Connor ◽

...

Keyword(s):

Herpes Simplex Virus ◽

Herpes Simplex ◽

Viral Replication ◽

Morphological Changes ◽

Reporter Virus ◽

Infection Cycle ◽

Herpes Simplex Virus 1 ◽

Fluorescent Reporter ◽

Simplex Virus ◽

The Impact

AbstractHerpesviruses are large and complex viruses, and have a long evolutionary history with their host species. One poorly understood, but important, area in virus-host interaction is the extensive remodelling of intracellular organelles and morphology that occurs during active viral replication. We have constructed a recombinant reporter virus that allowed us to classify four distinct stages in the infection cycle of herpes simplex virus 1. Single-cell analysis of the impact of the viral load on replication dynamics demonstrates the utility of this times-tamping method for tracking the infection cycle. High-resolution microscopy analysis of cellular structures in live and fixed cells in concert with our dual-fluorescent reporter virus have enabled us to generate a detailed overview over the spatial and temporal organisation of the cytoskeleton and organelles during viral replication. This provides the first systems-level analysis of the morphological changes involved in viral replication.

Download Full-text