Scientific Program

Conference Series Ltd invites all the participants across the globe to attend 10th International Virology Summit Vienna, Austria.

Day 2 :

Keynote Forum

Bryan R Cullen

Duke University Medical Center, USA

Keynote: Viral epitranscriptomics

Time : 09:30-10:00

Conference Series Euro Virology 2018 International Conference Keynote Speaker Bryan R Cullen photo
Biography:

Bryan R. Cullen obtained a B.Sc. in Biochemistry from Warwick University in the UK and a M.Sc. in Virology from the University of Birmingham before moving to the USA, where he obtained a Ph.D. in Microbiology from Rutgers University. In 1987, he was recruited to Duke University Medical Center as a Howard Hughes Medical Institute Investigator. He currently holds a James B. Duke Professorship in the Department of Molecular Genetics and Microbiology at Duke. Dr. Cullen’s research interests have historically revolved around the use of viruses as genetic tools to understand aspects of the biology of the eukaryotic cell, focusing particularly on RNA-sequence mediated gene regulation. Currently, his laboratory is studying the regulation of viral mRNA expression by viral microRNAs and by epitranscriptomic modifications. Dr. Cullen has published over 315 research papers and is on the editorial board of 11 prominent journals.

 

Abstract:

While it has been known for almost 40 years that a wide range of virally encoded RNAs, including mRNAs, are extensively modified by addition of a methyl group to the N6 position of adenosine (m6A), the functional consequences of this epitranscriptomic modification had remained unclear. However, the prevalence of m6A across multiple different viral species, and the recently demonstrated conservation of m6A residues in several distinct isolates of HIV-1, clearly implies that m6A favors some aspect of the viral replication cycle. More recently, the identification of the factors that add m6A to mRNAs, especially the m6A “writer” METTL3, and the definition of factors that bind m6A on mRNAs, including the key m6A “reader” YTHDF2, have recently allowed the phenotypic consequences of m6A addition to be more fully characterized. Using overexpression and/or genetic ablation strategies, as well as by mapping and mutationally inactivating specific viral mRNA m6A addition sites, we have now examined the effect of m6A in the context of three distinct viral families: the lentivirus HIV-1, the paramyxovirus influenza A virus (IAV) and the polyomavirus SV40. In all three cases, we observe that addition of m6A strongly enhances viral mRNA and protein expression, and hence replication, and, in the case of IAV, increases pathogenicity in vivo. In this presentation, I will discuss our current understanding of the mechanistic basis for this phenomenon.

 

Keynote Forum

Adrian Whitehouse

University of Leeds, United Kingdom

Keynote: Targeting herpesvirus ribonucleoprotein particle assembly: A novel antiviral strategy

Time : 9:00 - 11:15

Conference Series Euro Virology 2018 International Conference Keynote Speaker Adrian Whitehouse photo
Biography:

Adrian Whitehouse obtained a BSc in Microbiology from the University of Sheffield in 1991 and DPhil in Molecular Virology from the University of Oxford in 1994. Following Postdoctoral work at the Molecular Medicine Unit, St James’s Hospital in Leeds, he was awarded a Medical Research Council Non-clinical Fellowship in 1998, and joined the School of Molecular & Cellular Biology, University of Leeds, as a Lecturer in 2002, and subsequently appointed to Reader in 2005 and Professor of Molecular Virology in 2010. His research focuses on investigating the virus-host cell interactions which regulate the replication and transformation of human tumour viruses.

 

Abstract:

Human herpesviruses are responsible for a range of debilitating acute and recurrent diseases, including a number of malignancies. Current treatments are limited to targeting the herpesvirus DNA polymerases, but with emerging viral resistance and little efficacy against the oncogenic herpesviruses, there is an urgent need for new antiviral strategies. Here, I will describe a mechanism to inhibit the replication of the oncogenic herpesvirus Kaposi’s sarcoma associated herpesvirus (KSHV), by targeting the ATP-dependent formation of viral ribonucleoprotein particles (vRNPs). I will demonstrate that small-molecule inhibitors that selectively inhibit the ATPase activity of the cellular human transcription/export complex (hTREX) protein UAP56 result in effective inhibition of vRNP formation, viral lytic replication and infectious virion production. Strikingly, as all human herpesviruses use conserved mRNA processing pathways involving hTREX components, we demonstrate the feasibility of this approach for pan-herpesvirus inhibition.

 

Keynote Forum

Taijiao Jiang

Institute of Basic Medical Sciences-CAMS, China

Keynote: Bioinformatic approaches to Infectious Diseases

Time : 10:00-16:30

Conference Series Euro Virology 2018 International Conference Keynote Speaker Taijiao Jiang photo
Biography:

Taijiao Jiang has completed his PhD in Biochemistry and Molecular Biology at the Institute of Biochemistry and Cell Biology-Chinese Academy of Sciences in 1999 and his Master Degree in Computer Science from Yale University in 2003. After his Postdoc researches in Yale Biology Department (1999-2002) and MIT Biology Department (2003-2005), he got a Professorship at Institute of Biophysics Chinese Academy of Sciences. In 2015, he joined at the Chinese Academy of Medical Sciences & Peking Union Medical College and cofounded the Suzhou Institute of Systems Medicine. He is the Director of Center of Biomedical Big Data of Chinese Academy of Medical Sciences. He has published more than 40 papers in reputed journals and has been serving as the President of Bioinformatics Branch of the Chinese Biophysics Society since 2017.

Abstract:

Due to the development of high throughput sequencing technologies, large-scale sequencing of pathogens, such as influenza viruses and HIV, has not only become a routine work in surveillance of infectious diseases, but also become indispensable in identification of newly emerging infectious disease in their outbreaks. Besides, analyses of these large-scale sequence data of pathogens will significantly enhance our understanding towards the evolution of diseases, promoting new strategies for prevention and control of these viruses. His research mainly focuses on modeling the evolution of infectious disease from gene data and epidemiological data. Many computational methods have been developed in his lab to understand the origin and evolution of influenza viruses. They were demonstrated to be helpful in influenza vaccine recommendation, estimating the mortality burden and disease lethality, sourcing newly emerging influenza viruses, and so on. They have made a significant influence in the field of bioinformatics and informatics of infectious diseases. Among them, in collaboration with China CDC, the new methods developed for vaccine recommendation of human influenza viruses have now been in use in China CDC for assisting flu vaccine strains selection. 

  • Speaker Session
Location: vienna
Speaker
Biography:

 

Amina Khatun has completed her PhD in Veterinary Medicine (Microbiology) from Chonbuk National University (CBNU), South Korea. Currently, she is working as a Postdoctoral Researcher in the Department of Veterinary Immunology, College of Veterinary Medicine, CBNU. Her major research mostly focuses on functional genomics of PRRSV largely prospecting of viral pathogenesis to address two major hurdles in PRRSV vaccinology i.e. low genetic fidelity and limited cross-protection. Previously, she has completed her DVM and MS (Veterinary Pathology) from SAU, Sylhet, Bangladesh. She has published more than 10 papers in reputed journals. She has also been serving as a Reviewer of Ecology and Evolution Journal.

 

 

Abstract:

Low genetic fidelity and limited cross-protection are two major hurdles faced in porcine reproductive and respiratory syndrome virus (PRRSV) vaccinology, which is the most challenging threat to the swine industries worldwide. MLV vaccines are commonly used for homologous protection though there have been safety concerns as vaccine viruses reverted to virulence reported in fields. RVRp22, a ribavirin-resistant attenuated phenotype of PRRSV had high genetic stability during sequential replication in pigs reported in previous study. Wherein, ORF1a of RVRp22 genome was assumed to be involved in enhanced genetic stability and viral virulence. Therefore, the present study constructed four chimeric viruses based on ORF1a, 1b, 1ab and nsp2 regions of RVRp22 genome named as RVRp22-1a, 1b, 1ab and nsp2, respectively into VR2332  backbone to see the potential role of ORF1a in viral virulence and genome stability during serial passages in pigs following in vitro evaluation in MARC-145 cells and PAMs. In results, the replication of RVRp22-1a, 1ab and nsp2 was significantly suppressed in PAMs but replicated efficiently in MARC-145 like RVRp22 (attenuated strain). RVRp22-1b replicated in both of PAMs and MARC-145 cells with the similar trend as found in VR2332 (prototype strain). Consistently, RVRp22-1a, 1ab and nsp2 challenged pigs had lower viral loads in sera and lungs than RVRp22-1b, which was maintained even after third passage. Furthermore, RVRp22-1a, 1ab and nsp2 showed significantly a lower mutation frequency in nsp2 (most variable region in PRRSV genome) compared to RVRp22-1b. In conclusion, the present study indicated that ORF1a of RVRp22 genome might be critically involved in PRRSV virulence and the attenuated replication was maintained during serial passages in pigs which were enhanced with high genetic stability.

 

Biography:

Remziye Nalcacioglu has completed her PhD in 2003 from Karadeniz Technical University, in Turkey. She has been working in the same universirty as an academician. She is working on insect viruses including baculoviruses, iridoviruses.
 

 

Abstract:

A broad survey of the fall webworm, Hyphantria cunea (Lepidoptera: Arctiidae) populations in agricultural and forested areas in the central black sea region of Turkey led to the detection of granuloviruses (GVs). Thirty insect cadavers were collected from different locations, 8 of which contained GVs. All of them were determined to be Hyphantria cunea granulovirus (HycuGV) by tissue PCR and sequence analysis. A selected isolate (Hc1) was characterized and tested against third instar larvae of H. cunea. Electron microscopy confirmed typical GV morphology with ovoid granules of approximately 318-546 nm × 174-240 nm. Each granule contained a single rod-shaped virion with a mean size of 35-51 nm × 202-341 nm. The genome was estimated to be ~123 kb by restriction endonuclease analysis. Partial sequencing of the granulin ,late expression factor-8 (lef-8) and late expression factor-9 (lef-9) genes also confirmed the identity of the virus as HycuGV. A phylogenetic analysis based on these conserved genes, HycuGV-Hc1 grouped together with the previous HycuGV isolate (A5-1) and Estigmene acrea granulovirus (EsacGV) isolate from the same family. The LC50 of Hc1 isolate was 2.6×104 occlusion bodies (OBs/ml) against third instar H. cunea larvae. HycuGV-Hc1 caused 80.17% mortality with 109 OBs/ml in pot experiments performed in a greenhouse. This is the first study to describe a novel Turkish HycuGV-Hc1 isolate and preliminary data suggest the virus have a significant potential as an effective biopesticide for H. cunea control.

 

Yael Fridmann Sirkis,

Weizmann Institute of Science, Israel

Title: Mimivirus on a bad hair day
Speaker
Biography:

Yael Fridmann Sirkis has her expertise in Structural Biology as well as characterizing protein-protein interactions. She is currently working in the protein analysis unit at the Weizmann Institute of Science in Israel.

 

Abstract:

Acanthamoeba polyphaga mimivirus (APMV) is the first giant virus discovered almost 15 years ago. It has uncommon characteristics such as a stargate shape in one of its vertices through which its dsDNA is released into the cytoplasm. It also has an external thick fibril layer that was shown to be important for adhesion. This finding was based on mimivirus strain that suffered a drastic reduction in the number of its fibrils after a third of its genes lost activity during continuous passaging in germ-free amoebae (PMID:21646533). Here, we subcultured mimiviruses under normal conditions and continually passed them through 0.45 mm filter. Thus, a population of hair deficient (HD) viruses was enriched and particles were cloned and imaged. Genomic analysis of the filtered viruses revealed three mutations that affected only three genes. One of the mutations showed an in-frame deletion in L71 gene, a collagen-like protein that eliminated almost all of its collagen motif sequences. The resulting HD viruses revealed a significant reduction in their infection titer as well as substantially reduced virus yield. HD-infected amoebae also burst less readily.

 

Speaker
Biography:

 

Francielle Tramontini Gomes de Sousa has her expertise on dengue pathogenesis and evaluation of antiviral activity and mechanism of action of natural and semi-synthetic compounds. She has developed an in vitro co-culture model of endothelial cells and monocytes to screen compounds with therapeutic potential for management of endothelial extravasation in severe dengue. In a previous work, she had shown that serum from acute severe dengue patients differentially affects endothelial cells barrier function in vitro and found some correlations between immunomediators and vascular leakage. Her current interests include studying the pathogenic mechanism of dengue vascular leak as well as searching for therapeutic candidates against dengue virus by using in vitro (endothelial cells) and in vivo (murine) models.

 

Abstract:

Introduction: Vascular leakage is an adverse outcome in response to dengue virus (DENV) infection, resulting in depleted intravascular volume and hypotension, which may evolve into hypovolemic shock. Dengue non-structural protein 1 (DENV NS1) has been shown to contribute to pathogenesis by directly triggering endothelial glycocalyx layer (EGL) degeneration, in a cytokine independent pathway, as well as by inducing the release of vasoactive cytokines from PBMCs, both leading to plasma leakage. Agaricus brasiliensis is a basidiomycete fungus native to Brazil widely consumed and studied due its therapeutic properties, most of them related to its polysaccharidic content. A (1→6)-(1→3)-beta-D-glucan isolated from A. brasiliensis fruiting bodies (FR) was chemically modified to produce its corresponding sulfated derivative (FR-S). Since there is no specific treatment for DENV and the current available Sanofi dengue vaccine does not confer full protection to the disease, reversing or preventing EGL degeneration has therapeutic potential on severe dengue.

 

Methodology & Theoretical Orientation: FR-S was tested against DENV NS-1 induced endothelial barrier disruption by measuring trans-endothelial electrical resistance (TEER). The effect of FR-S on NS-1 binding to endothelial cells was evaluated by immunohistochemistry.

 

Findings: Figure 1 show that FR-S completely inhibited TEER reduction induced by DENV-2 NS1 treatment on HPMECs at the two higher concentrations tested (0.25 and 0.12 μg/mL). At the two lower tested concentrations (0.06 and 0.03 μg/mL), no protection against NS1-induced TEER reduction was observed. Results in figure 2 show that the mechanism by which FR-S prevents endothelial barrier dysfunction includes inhibition of NS1 binding to HPMECs at a concentration dependent manner.

 

Conclusion & Significance: The findings indicate that FR-S inhibits NS1 binding to endothelial cells and prevents NS1 induced endothelial dysfunction. FR-S may have an anti-vascular leak effect since NS1 is in part responsible for the plasma leakage occurring in patients with severe dengue.

 

Speaker
Biography:

Tae Jin Choi has completed his PhD from University of California, Berkeley in 1993 and 2 years Postdoctoral studies from University of Wisconsin, Madison. Since 1995 he is working as a Professor in Pukyong National University in Busan Korea. He had published over 70 papers on the viruses of plant, fish, shrimp and vaccines for aquatic viruses. Recently he is working on the development of microalgae as bioreactor for recombinant protein production and bacteriophage biocontrol of water melon.

 

Abstract:

Statement of Problems: Bacterial fruit blotch (BFB) is an economically important bacterial disease that has caused huge economic losses in melon and watermelon crops around the world. There is no commercially available cultivars resistance to this disease caused by bacteria Acidovorax citrulli. Mainly, two genotypes (genotype I and II) are reported in A. citrulli, in which genotype II is the main causal agent of BFB in water melon that is major problem in Korea.

 

Methodology & Theoretical Orientation: We have isolated more than 50 bacteriophages infecting A. citrulli, from watermelon leaf samples which were collected from different parts of Korea. Two of the isolated phages with large plaque size named as ACP17 and ACPHW were further characterized and used for phage biocontrol.

 

Findings: Based on electron microscope observations, ACP17 belongs to Myoviridae family with head diameter 100±5 nm and a tail length of 150±5 nm while ACPHW has a head size of 60±5 nm and tail size 180±5 nm which belongs to Siphoviridae family. Among forty A. citrulli strains, ACP17 can lyse 27 strains of which most belongs to genotype I, and ACPHW can lyse 39 strains containing group I and II. In planta assay showed that the germination rate of watermelon seeds coated with the bacteriophages was up to 80% in the presence of A. citrulli contrast to untreated seed showing no germination. Also, these germinated plants showed 100% survival in A. citrulli treated soil.

Conclusion & Significance: These results suggest the possible use of these phages as an effective bio control agent for BFB.

 

Speaker
Biography:

Henry Memczak studied nanotechnology at the University of Kassel, Germany and completed his PhD in biochemistry in 2014 at the University of Potsdam, Germany and the Fraunhofer Institute for Cell Therapy and Immunology, Germany. He has worked on the development of analytical biosensors for influenza detection and methods for peptide-based biointeraction analysis for several years, published several papers, holds two patents and co-founded the company qpa bioanalytics GmbH for the commercialization of novel peptide biochips. For his dedicated translational research he received several awards and scholarships.

 

Abstract:

The only cost-effective protection against influenza is vaccination. Due to rapid mutation continuously new subtypes appear, what requires annual reimmunization. For a correct vaccination recommendation, the circulating influenza strains have to be detected promptly and exactly and characterized regarding their antigenic properties. Due to recurring incidents of vaccine mismatches, there is a great need to speed up the process chain from identifying the right vaccine strains to their administration. The monitoring of subtypes as part of this process chain is carried out by national reference laboratories within the WHO Global Influenza Surveillance and Response System (GISRS). To this end, thousands of viruses from patient samples (e.g. throat smears) are isolated and analyzed each year. Currently this analysis involves complex and time-intensive (several weeks) animal experiments to produce specific hyperimmune sera in ferrets, which are necessary for the determination of the antigen profiles of circulating virus strains. These tests also bear difficulties in standardization and reproducibility, which restricts the significance of the results.

To replace this test a peptide-based assay for influenza virus subtyping is developed. The differentiation of the viruses takes place by a set of specifically designed peptidic recognition molecules which interact differently with the different influenza virus subtypes. The differentiation of influenza subtypes is performed by pattern recognition guided by machine learning algorithms, without any animal experiments.

Speaker
Biography:

 

Diana Leibman completed her PhD in 2012 from the Hebrew University in Jerusalem. She works as a Research Engineer in the Agricultural Research Organization, The Volcani Center. The research topics in which she is involved are: The role of RNA-dependent RNA polymerase 1 in plant defense against viruses; CRISPR/Cas9 genome editing technology for crop improvement; development of resistance to RNA and DNA viruses in cucurbits and tomato by transgenic approaches; identification of plant genes associated with disease symptom development to plant virus infection and; genetic engineering of attenuated ZYMV-AG as a plant virus vector, for gene expression and epitope presentation. 

 

Abstract:

RNA-dependent RNA polymerases (RDR) play an important role in virus protection and plant gene control. Their activity is based on the synthesis of double stranded RNA that is a template for the gene silencing system. Four RDR1 (RDR1a, b, c1, c2) genes were identified in cucumber having different expression levels before and after infection by various viruses. The CsRDR1a, b, c genes have a homology of 60-58% at the amino acid level, while CsRDR1c1 and c2 are almost identical (97% homology), but have different promoter sequences. A high expression level of CsRDR1b was characterized in cucumber strains with partial resistance to a number of viruses, while CsRDR1c1 and c2 not expressed in healthy plants. Cucumber plants infected by several different viruses showed a moderate expression level of CsRDR1b and a dramatic level of CsRDR1c1, c2. The expression level of CsRDR1a, CsRDR2 and CsRDR6 did not increase following virus infection. The differential expression of CsRDR1b and CsRDR1c1, c2 indicates dissimilar control of these genes. The relationship between the high levels of expression of CsRDR1b in resistant cucumber cultivars reinforces the assumption that this gene is unique for virus resistance. Using the CRISPR/Cas9 system, we created a variety of mutants in the rdr1b and rdr1c1 and c2 genes. Knockdown of these genes increased virus accumulation. Plants with a homozygous mutation in the rdr1b genes (deletion of 34 nucleotides) and rdr1c (2 and 1 nucleotides in the rdr1c1 and rdr1c2 genes respectively) showed increased susceptibility to Cucumber mosaic virus 4 dpi, especially rdr1c mutants plants which collapse 6 days after infection. The viral symptoms increased also after Cucurbit vein yellowing virus infection, whereas the susceptibility to Zucchini yellow mosaic virus infection was less. Notably, CsRDR1b expression in healthy rdr1b mutants was less than in non-mutant plants. The control of CsRDR1b and CsRDR1c1, c2 is under study in our laboratory to help understand the silencing mechanism of plants defense.

 

Speaker
Biography:

Dr. Slobodan Paessler, is a Professor in the Department of Pathology and Director of Galveston National Laboratory Preclinical Studies Core. Dr. Paessler is a co-principle investigator on the Universal Influenza Vaccine project funded by an NIAID grant at Etubics Corporation. He serves as the Director of Animal Biosafety Level 3 for the Institute of Human Infections and Immunity. He has been Member of Scientific Advisory Board at Etubics Corporation since July 2015. He serves as a Member of the Center for Biodefense & Emerging Infectious Diseases. He received a Dr. Med Vet (D.V.M) at Ludwig-Maximilian University and a Ph.D in Experimental Pathology from UTMB.

Abstract:

Flu epidemics and potential pandemics pose great challenges to public health institutions, scientists and vaccine producers. Creating right vaccine composition for different parts of the world is not trivial and has been historically very problematic. This often resulted in decrease in vaccinations and reduced trust in public health officials. To improve future protection of population against flu we urgently need new methods for vaccine efficacy prediction and vaccine virus selection. Recently, novel bioinformatics platform based on electronic biology was successfully utilized for real-time monitoring of influenza A viruses as well as for prediction of vaccine efficacy in Australia and USA in 2017 and 2018. Here we present wEB platform and its usage in identifying functional biological changes in haemagglutinin protein of influenza A viruses and how this knowledge can be applied for vaccine design, prediction of future vaccine efficacy and real-time virus evolution monitoring.

 

Speaker
Biography:

Cha Gyun Shin has completed his PhD from The Ohio State University and Postdoctoral studies from Dana Faber Cancer Institute. He is a Professor at Department of Systems Biotechnology, Chung-Ang University, Republic of Korea.

 

 

Abstract:

Integrase is the retroviral protein responsible for incorporating a double-stranded viral DNA copy into the host chromosome. Single amino acid substitution in integrase (IN) active site is replication-defective, and reduces viral infectivity. This study is to investigate whether mutation in DD(35)E motif and residues near the active site of catalytic core domain is critical for feline foamy viral integration.  In vitro enzymatic activities of IN mutant proteins were analyzed by using p32-radiolabeled oligonucleotide substrates and 15% polyacrylamide gels. DDE mutation almost lost their IN activities. But Q165A, Y191A, and S195A showed reduced effects. Although DDE mutants produced replication-defective virions by one cycle transfection, Q165A, Y191A, and S19A mutants had infectivity on their natural host cells. Known as the immature virions containing mutated IN executed integration aberrantly and had trouble in producing viral DNAs for new virion particles, in the case of DDE mutants no integrated viral DNAs were detected at 24 h and 48 h post infected host chromosomal DNAs. However, integration of viral DNA whose virions have IN mutants in the amino acid residues present near the active site such as Q165A and Y191A were detected, and then quantitated by competitive PCR. Nonsynonymous substitutions in highly conserved region of feline foamy viral IN resulted in viruses with replication-defective. Defects in viral DNA synthesis, viral production, and integration processing were observed for all of the replication-defective mutants. Especially, feline foamy viral natural host cells are not infected with replication-defective DDE mutant viruses.

 

Speaker
Biography:

Dr. Kim has completed his PhD from the Department of Plant Pathology, North Carolina State University (NCSU) and postdoctoral studies from NCSU Department of Biochemistry. He is the director of Plant Clinic, Seoul National University. He has published more than 100 papers in reputed journals and has been serving as an editorial board member of Virology, Virus Research, and Scientific Reports.

 

Abstract:

Soybean mosaic virus, a member of the genus Potyvirus, significantly reduces soybean production worldwide. Rsv3, which confers strain-specific resistance to SMV, was previously mapped between the markers A519F/R and M3Satt in chromosome 14 of the soybean [Glycine max (L.) Merr.] genotype L29. Analysis of the soybean genome database revealed that five different NBS-LRR sequences exist between the flanking markers. Among these candidate Rsv3 genes, the full-length cDNA of the Glyma.14g204700 was successfully cloned from L29. Over-expression of Glyma.14g204700 in leaves inoculated with SMV inhibited viral infection in a soybean genotype lacking Rsv3. In addition, the transient silencing of the candidate gene caused a high accumulation of a virulent strain in L29 carrying Rsv3. Our results therefore provide additional line of evidence to support that Glyma.14g204700 is likely Rsv3 gene that confers strain-specific resistance to SMV.

 

 

Speaker
Biography:

Sherwin Morgan completed his respiratory care training from Malcolm X College of Respiratory Care in Chicago, IL. He is an advanced respiratory care practitioner with the National Board for Respiratory Care in the United States. He is Clinical Practice and Development /Educator/Research Coordinator for the Department of Respiratory Care Services, Section of Pulmonary and Critical Care Medicine at the University of Chicago Medicine. He has published more than 25 peer review papers in multiple medical journals. He has designed, engineered, and collaborated with a number of research studies with the pulmonary medicine department.

Abstract:

The mechanics of flu related respiratory illness is not completely implicit as it includes; influenza, zoonotic and non-influenza pathogens. Precise diagnosis is difficult as it often mimics asthma out of control which has perplexed researchers for decades. This has led to treatment confusion and an underestimation that the primary cause of breathing difficulties is related to bronchiolitis-bronchiectasis. A microbiology respiratory viral panel (RVP) test via polymerase chain reaction (PCR) can identify whether there is a co-existing viral lung infection that may worsen the lung function. Viral flu-related respiratory infections are highly transmittable and may increase morbidity and mortality in patients with premorbid pulmonary disease and weakened immune systems. The symptoms of flu include dyspnea and coughing; after usual treatment with steroids and asthma medications, continue to have worsening symptoms causing re-hospitalization. Chest radiography for patients with respiratory distress due to flu are notable for; bronchial wall thickening, bronchiectasis and sub-segmental atelectasis, related air-flow obstruction. Rhinoviruses (RV) – enterovirus (EV) for example is under recognized as the leading cause of hospitalization for viral outbreaks. Respiratory Enterovirus is responsible for 10 to 15 million hospitalizations annually. Enterovirus (D-68) was attributed to 14 deaths in 2014 in the United States (USA) and 70 deaths in the 2011 Philippines D68 outbreak. Ever since the 2014 D68 outbreak, there has been a drastic increase in the number of patients hospitalized and re-hospitalized for flu symptoms associated with severe acute respiratory distress on the pediatric and oncology wards. Zoonotic agents such as coronavirus (HCoV) are passed bi-directionally between animals and humans and capable of joining with other viral agents. All this has created undefined burden on global clinical resources. More research is needed to understand the pathogenesis of viral bronchiolitis and bronchiectasis related respiratory illness to assist clinicians with recognition and treatment of this highly morbid disease.

Speaker
Biography:

 

Christopher Chadwick is a Technical Officer in the Global Action Plan for Influenza Vaccines Secretariat at World Health Organization. Previously, he was a Global Health Officer in the office of Global Affairs at the US Department of Health and Human Services. He received a Master of Science degree in Public Health with a concentration in microbiology and emerging infectious diseases from Milken Institute School of Public Health at George Washington University and a Bachelor of Science in Microbiology from Louisiana State University.

 

Abstract:

Purpose: The World Health Organization’s Global Action Plan for Influenza Vaccine (GAP) was a 10-year initiative dedicated to reducing the global shortage and inequitable access to influenza vaccines in the event of an influenza pandemic. The overarching goal of the GAP was to develop the capacity to produce enough vaccines to immunize 70% of the global population with two doses of vaccines. The GAP aimed to achieve this goal by increasing evidence based seasonal influenza vaccine use; developing influenza vaccine production and regulatory capacity in 14 low and middle income countries (LMICs) and; encouraging the development of improved influenza vaccines.

 

Methods: Between 2006 and 2016, the WHO collaborated with member states and key stakeholders to address the global shortage of and increase equitable access to pandemic influenza vaccines in the event of an outbreak.

 

Results: The outcomes of the GAP include: A dramatic increase in countries with a seasonal influenza policy in place (115 member states by 2014 from a baseline of 74 in 2006); the development of 8 licensed pandemic influenza vaccines and 3 licensed seasonal influenza vaccines in 6 LMICs and; A global expansion of pandemic vaccine production capacity, especially in LMICs (potential global capacity of 6.4 billion doses estimated in 2015).

 

Discussion: Following the conclusion of the GAP in 2016, priorities for influenza vaccine preparedness moving forward are to sustain the production capacity of influenza manufacturers in LMICs, promote and stimulate innovative influenza vaccine research and development, identify root causes of influenza vaccine hesitancy, generate more evidence on vaccine effectiveness in specific risk groups, and identify innovative ways of addressing global pandemic influenza preparedness.

 

  • poster session
Location: vienna
Speaker
Biography:

Hyoun Sub Lim was trained for his PhD in University of Illinois at Urbana-Champaign and he continued Postdoctoral studies in University of California at Berkeley. His researches have mainly focused on plant viral movement in plant cell for more than 20 years and more than fifty published papers proved his field in Plant Virology. Currently he is a Professor in Chungnam National University, Korea and has worked as an Editorial Board Member of Plant Pathology journal.
 

 

Abstract:

In Asia, Chinese cabbage (Brassica rapa) is mainly grown and utilized in various ways as a healthful food source. However, increasing virus damage resulting from changes in trade of agriculture products including seedlings and seeds, as well as climate change and repeated cultivation, has reduced Chinese cabbage production. According to recent research, three plant viruses - Turnip mosaic virus (TuMV), Cucumber mosaic virus (CMV), and Youcai mosaic virus (YoMV) - are reported to affect Chinese cabbage yields. Recently we detected new isolates of YoMV in Korean radish fields, and full-length infectious clones of two isolates were generated in the dual 35S /T7 promoter driven pJY binary vector. Four amino acid differences (V383I, M492I in 125kDa, T1245M in 182kDa and M17T in CP) between two isolates resulted in either severe or mild symptom development in Nicotiana benthamiana. In order to reveal the amino acids related to severe pathogenesis, four hybrid constructs were generated through by gene exchanges between the isolates. Hybrid constructs maintaining CP residue 17 as threonine in the MP/CP overlap region developed severe symptoms. Further analysis expressing CPM(17)T from a Potato virus X vector produced differential symptoms in N. tabacum cv. Xanthi, inducing HR (T17) and mild symptoms (M17) respectively.

 

Speaker
Biography:

Mu Kuan Chen is currently the Superintendent of Changhua Christian Hospital. He is a prolific author in Head and Neck Medicine. He has served on numerous Editorial Boards of several international journals, been invited to be the book chapter writer. His main interests are focused on development of new techniques of minimally invasive surgery in otorhinolaryngology, head and surgery including endoscopic skull base surgery, endoscopic nasopharyngectomy, endoscope assisted parotidectomy and ablation of submandibular gland, and endoscopic sinonasal tumor excision. Besides, he has published many articles which focused on basic research of oral cancer and nasopharyngeal cancer

Abstract:

Oral squamous cell carcinoma (OSCC) is the most common primary malignancy occurring in the head and neck. It is the fourth most common male cancer and the fourth leading cause of male cancer death in Taiwan. Metastatic tumors are extremely common in the late stages of cancer. Treatment for metastatic cancer aims to decelerate the growth or spread of cancer. The extracellular matrix degradation involves various proteases such as matrix metalloproteinase (MMPs), are calcium-dependent zinc-containing endopeptidases. MMP-2 is type-IV collagenases of MMPs, and correlated with lymph node metastasis and advanced tumor stage groups. Pinosylvin, a preinfectious stilbenoid toxin, is used to study its properties as a fungi toxin and therapeutic agent. Pinosylvin is used as a representative stilbene to study its biological actions and therapeutic value in processes such as cell survival, apoptosis and cell mobility. However, the pharmacological activities of pinosylvin in anti-metastasis remain unclear. In this study, we used wound closure assay and transwell assay to determine the effects of pinosylvin on oral cancer cell migration and invasion. Pinosylvin treatment significantly inhibited the migration and invasion abilities of oral cancer cells in vitro. Gelatin zymography results revealed that pinosylvin inhibited MMP-2 activity. In addition, pinosylvin suppressed carcinoma-associated epithelial–mesenchymal transition in oral cancer cells. Pinosylvin inhibits the invasion of human oral cancer cells and is a potential chemopreventive agent against oral cancer metastasis.

 

 

Speaker
Biography:

Boyoun Moon completed her phD thesis in college of veterinary medicine of Seoul National Univeirsity. She is charge of the diagnosis and prevention of companion animal viral disesases in animal disease diagnostic division(ADDD), Animal and plant qurantine agency (APQA) in South Korea.

 

Abstract:

Canine parvovirus2 (CPV2) is an etiological agent that causes acute hemorrhagic enteritis and fatal myocarditis in carnivore species. CPV2 evolved from feline parvoviurs by host transmission and has contineously been replaced by the genetic variants of capsid protein (VP2). In this study, it was investigated that the genetic diversity and phylogenetic relatedness of the VP2 gene encoding capsid protein in CPV2 obtained from symptomatic domestic dogs obtained ranging from 2011 to 2017 in Korea. Positively selected sites on VP2 of CPV2 variants during a decade was identified and the multiple proteins were evaulated with the theoretical homology modeling using SWISS-MODEL and PyMOL. To investigate the subtype of CPV circulating in Korea, a total of 28 partial VP2 sequences were analyzed. Both CPV2a (15/28, 53.6%), and  CPV2b (12/28, 42.9%) were dominant subtypes and Korea CPV2c in 2017 was first detected in this study. Of them, eighteen complete VP2 nucleotide sequences (1755bp) and the amino acid were constructed with phylogenetic tree. A clustering of our variants was closely related to China and Vietnam strains, but in different groups from US, Eruope and vaccine strains.Non-synonymous mutations, Phe267Ala, Tyr324Ile and Thr440Ala have been increased during last decade and minor 10 mutations have been also observed. A codon-based maximum likelihood analysis of dN/dS (ω) revealed that specific amino acid 324 (ω=3.518), 426 (ω=3.517) and 440 (ω=3.459) in VP2 gene was under strong positive selection of Korea CPV2. These results support that dynamic process of CPV2 in Korea results in the locally adaptation.

 

Biography:

Abstract:

Introduction: Hepatitis C virus (HCV) replication is closely linked to lipid metabolism. Therefore, lipidomic analysis of HCV infected hepatic cells can offer insights into the pathogenesis of HCV infection and identify molecular targets that can serve as potential targets for new treatments.

 

Aim: The aim of this project was to study the effects of HCV infection on intracellular lipid homeostasis and turnover in hepatic cells. We have previously reported HCV-induced changes in neutral lipids; so now present data on phospholipids.

 

Methods: Huh7 cells were infected with HCV (JFH1 strain) and cultured until they were 90% infected. Cellular lipids were separated by high performance thin layer chromatography (HP-TLC) to measure changes in the major phospholipid species. The rate limiting enzymes of phosphatidylcholine metabolism were knocked down and the effects on HCV replication were measured. 

 

Results: HP-TLC showed increased amounts of phosphatidylcholine in lipid extracts from whole cells and from ER fractions of JFH1 infected Huh-7 cells. PC was the only phospholipid species detected in purified lipid droplets, and was significantly increased in infected cells. PYTC1 (CTP: phosphocholine cytidylyltransferase) and PEMT (phosphatidylethanolamine N-methyltransferase) are the rate limiting enzymes of PC biosynthesis in hepatocytes. Silencing PYTC1 had no effect on HCV replication or infectivity. However, when PEMT was silenced, both viral replication and infectivity were decreased by more than 50%, and less lipids accumulation was observed.

 

Conclusion: Our previous data reveal global changes in lipid abundance, particularly in the ER, which are predicted to impact the HCV life cycle and pathogenesis. We now report increased PC content in the ER and in lipid droplets. Our data suggest that in HCV infected cells the minor PC synthesis pathway is most important, as inhibiting PEMT inhibits replication and production of infectious virus.

 

Speaker
Biography:

Shu Hui Lin has her expertise in gene mutation test, cancer diagnosis and cancer research. She is a Leader of Molecular Pathology Department of Changhua Christian Hospital in Taiwan. She has PhD of Institute of Medicine. She is committed to cytological diagnosis and establishment technical of gene mutations test. Her research field focuses on oral squamous cell carcinoma and she hopes that she can find novel therapeutic targets in oral cancer.

 

 

Abstract:

Background: Oral cancer is the fourth highest incidence of malignancy in males and the seventh highest in the general population of Taiwan. Oral squamous cell carcinoma (OSCC) is the main subtype of oral cancer, which accounts for >95% of all cases of oral cancer. Discoidin domain receptors (DDRs), a collagen receptor tyrosine kinase, play a major role in cancer progression. DDR2 has been suggested as a prognostic marker in several cancer types; however, the correlation between DDR2 expression and clinical outcome of oral cancer patients in Taiwan population has not been investigated.

 

Material & Methods: In the present study we sought to determine the clinical significance of discoidin domain receptor tyrosine kinase 2 (DDR2) expression in OSCC patients. We examined DDR2 expression in OSCC specimens by immunohistochemistry and then we analyzed the association of DDR2 expression with clinicopathological factors in OSCC.

 

Result: We divided 254 OSCC cases into two groups based on DDR2 expression levels and compared with several clinicopathological factors and their overall survival. The group with high DDR2 expression had significantly higher frequencies of lymph node metastasis (P=0.0094) and AJCC stage (P=0.0058) compared to the group with low DDR2 expression. Furthermore, the lymph node metastasis oral cancer patients with high DDR2 expression had low survival rate than low DDR2 group (P=0.0458).

 

Conclusions: Our data indicate that DDR2 is a potent biomarker that can be used as an effective therapeutic target for treating OSCC patients with lymph node metastasis.

 

Speaker
Biography:

In Sook Cho has completed her PhD degree of Plant Pathology from Chungnam National University, South Korea. Her researches have mainly focused on viral diseases of fruit trees for more than 10 years and more than forty published papers in Plant Virology. Currently she has worked as a Research Plant Virologist in National Institute of Horticultural and Herbal Science, Rural Development Administration, South Korea.
 

 

Abstract:

Apple is an economically important fruit crop, covering an area of 23,355 ha with an annual production of about 545,000 tonnes in South Korea. Apple trees are affected by at least six viruses and viroid diseases, which cause economic losses depend on the plant species and virus strains. Among these viruses, Apple chlorotic leafspot virus (ACLSV), Apple stem grooving virus (ASGV) and Apple stem pitting virus (APSV) commonly occur in Korean commercial apple orchards. In 2017, we observed several apple trees (Malus domestica Borkh.)  showing mosaic patterns on leaves cv. Shinano sweet or dappling and crinkling on fruits cv. Gamhong. To apply next generation sequencing (NGS) for apple virome of symptomatic apple trees, total RNA was extracted and used for constructing a cDNA library followed by high throughput sequencing (HTS). From the HTS data, ACLSV, ASGV, ASPV and Apple scar skin viroid (ASSVd) were identified, all of which have been previously reported in South Korea. In addition, Apple necrotic mosaic virus (ApNMV) recently described a novel ilarvirus was identified. To confirm the presence of ApNMV, RT-PCR was performed using specific primer pairs ApNMV-F and ApNMV-R. The sequence showed 94% identity with the CP region of the previously published ApNMV (LC108995) from the samples showing leaf mosaic symptoms. The ApNMV positive apple trees were also co-infected with ACLSV, ASGV and ASPV. The symptomatic fruit samples showing dappling and crinkling were infected with ACLSV, ASGV, ASPV and ASSVd. ASSVd is recognized as the causal agent of the dapple apple disease. ASSVd showed 99% nt. sequence identity with apple reference variants. NGS was a valuable and powerful tool for detection and identification of known and unknown virome in infected apple trees.

 

 

Speaker
Biography:

Hyoun Sub Lim was trained for his PhD in University of Illinois at Urbana-Champaign and he continued Postdoctoral studies in University of California at Berkeley. His researches have mainly focused on plant viral movement in plant cell for more than 20 years and more than fifty published papers proved his field in Plant Virology. Currently he is Professor in Chungnam National University, Korea and has worked for an Editorial Board Member of Plant Pathology journal.

 

Abstract:

As trading of agricultural products with neighboring countries has increased new plant diseases have been reported in Korea. Specifically, virus-contaminated imported seeds have damaged vegetable and fruit production. Little seed is now produced in Korea, and it is therefore very important to ensure that imported seed is not a source of new viruses. In order to investigate seed transmitted viruses we surveyed pepper fields nationwide and detected Pepper mild mottle virus (PMMoV) in the main pepper production regions of Sangcheng and Jeongson. We have generated full length clones of two isolates, named Sangcheong 47 (S-47, KX399390) and Jeongsong 76 (J-76, KX399389) respectively, in a T7 promoter-driven vector; sequencing revealed that these isolates shared ~99% nucleotide and amino acid identity, and are closely related to Japanese and Chinese isolates at the nucleotide level. Amino acid sequence comparisons revealed 99.73, 99.81, 98.44, and 100% identity in ORF1, ORF2, MP, and CP, respectively, between S-47 and J-76. Both isolates induced severe symptoms in Nicotiana benthamiana, but mild symptoms in Capsicum annuum. Each ORF was expressed as a GFP fusion from a binary vector, and no differences in subcellular localization were detected except for the 126 kDa proteins; the J-76 126 kDa clearly formed intracellular aggregates not observed with S-47 126 kDa protein. In addition, seed transmission rates from J-76 and S-47 infected Capsicum annuum showed 66% and 34% germinated plants from 500 harvested seeds respectively. Despite indistinguishable symptoms, J-76 showed a two times higher seed transmission rate that may result from some of the substitutions (S-47>J-76) of R(142)K, D(583)N, and V(931)I in 126 kDa and K(134)R, V(192)A, N(226)D, L(250)S in MP function with respect to S-47 and J-76.

 

 

Speaker
Biography:

Dóra Tombácz has completed her MSc in Biology (2006) and PhD in Medical Sciences (2010) from the University of Szeged, Hungary. She is working in the Department of Medical Biology as an Assistant Professor at the same university in the Genomics & Gene Technology group. She has published more than 30 papers in reputed journals. Her primary field of interest is transcriptomics, the analysis of different organisms at the RNA level. She is currently working with next- and 3rd generation sequencing techniques at the University of Szeged, and as a Visiting Assistant Professor at the Stanford University, USA.

 

Abstract:

High-throughput sequencing methods have revolutionized genomics, including RNA-sequencing; however the widespread short read-sequencing techniques cannot distinguish between the different transcript isoforms and overlaps. The third-generation, long-read sequencers (TGS) are enabled to identify full-length isoforms. For the deep analysis herpesvirus transcriptomes, we utilized the Pacific Biosciences (PacBio) RSII and the Sequel TGS machines, as well as the Oxford Nanopore Technologies (ONT) MinION device. In this project, we characterized the RNA profiles of the following viruses: human herpesvirus-1 (HSV-1), pseudorabies (PRV), varicella zoster virus (VZV), human cytomegalovirus (HCMV), Epstein-Barr virus (EBV). For the analysis of the static transcriptome of HSV, PRV and HCMV the PacBio IsoSeq protocol was used. For the analysis of the dynamic properties of PRV transcript isoforms we applied the PacBio non-amplified method. To obtain a more comprehensive picture about the RNA variants, the ONT cDNA and direct RNA sequencing protocols, as well as a CAP-selection method combined with ONT cDNA sequencing were also utilized. These ONT approaches were applied for the analysis of EBV and VZV transcriptomes. Our surveys redefined the herpesvirus transcriptomes profiles. We discovered that the herpesvirus genomes are much more complex and the transcriptional-read-throughs between the genes are more common than it was known. Our data revealed that these read-throughs can generate very long RNAs including some containing oppositely oriented ORFs, which have been classified as complex transcripts. The applied TGS techniques have been shown that the extensive RNA variations are common among herpesviruses

Hyun Kim

The Catholic University of Korea, South Korea

Title: Whole-genome sequencing analysis of SFTSV detected in South Korea
Speaker
Biography:

Hyun Kim has a Master's course from Catholic University School of Medicine. Mainly her study is waterborne viruses.

 

Abstract:

Severe fever with thrombocytopenia syndrome (SFTS) is a newly emerging infectious disease and caused by SFTS virus (SFTSV), a tick-borne phlebovirus in family Bunyaviridae. SFTSV was reported in China in 2011, more recently in Japan and South Korea. In this study, samples were collected from patients with SFTS in Jeju, South Korea in 2017, and its identity was confirmed as SFTSV by RT-PCR, and nucleotide sequencing and alignment analysis. The whole-genome of the SFTSV strains sequence analysis revealed three segments comprising the whole genome: L segment (6,368 bp), M segment (3,378 bp), and S segment (1,744 bp). The SFTS strain showed amino acid identities with Korea reference strains of 98.30-99.83%, 98.51-99.44%, and 99.38-99.57% in the S segment, M segment, and L segment, respectively. Additional, it showed 98.94-99.49%, 98.42-98.70%, and 99.09-99.42% similarity with Japan reference strains and similarities with reference strains in China were 98.39-99.28%, 97.49-98.23%, and 99.14-99.47%. Of the total amino acids, 28 mutations were identified, among 11 of which was special substitution mutations found only in our strains. This research will be used as a full-length SFTSV sequence standard for future comparison studies. Also, it may prove useful to the field of public health by facilitating the diagnosis and the prediction of new emerging variants.

 

 

Speaker
Biography:

In Hyuk Baek studied Biotechnology in Saarland University in Saarbrücken, Germany.  Currently, he is doing his PhD course under supervision of Prof. Dr. Helms and working with Dr. Youngjun Kim in the environmental safety group in the Kist-Europe (Korea Institute of Science and Technology Europe branch). His main topic of his Master’s thesis is metagenomic analysis of the viral communities in human feces: molecular approaches to discovery and characterization of novel viruses. He has also participated in several topics followings with 3 SCI (+1 submitted), 3 SCIE papers, 2 presentations and 4 registered patents.

 

 

Abstract:

Background: Combined 3D cell culture in vitro assay with microenvironments mimicking systems are effective for cell based drug and chemicals toxicity screening test to close tissue mimicking micro-environmental systems that was contributed in single or multiple compounds to check cell migration, angiogenesis, metastasis and morphogenesis. The angiogenesis structure of endothelial cells in the ECM was confirmed as 3D and regulated by surface treatment of the microfluidic channels. Filamentous bacteriophages are a member of the family Inoviridae. These are used for material science, drug delivery, tissue engineering, energy and biosensor. Genetically modified bacteriophages could deliver therapeutic molecules or genes to specific cancer tissues or organs.

 

Objectives: In this research, multi-functionalized biocompatible bacteriophage and ECM were applied for mimicking angiogenic micro-environmental systems.

 

Materials & Methods: E. coli MC1061 and K91BK were used for plasmid and phage amplification respectively. Phage preparation was performed by PEG/NaCl precipitation. The function of bacteriophages was tested by RT-qPCR, ELISA assay. The angiogenesis factors (e.g. CD31 and VE-cadherin) were confirmed by using confocal microscopy on commercialized lab-on-a-chip.

 

Conclusion: In this study, we demonstrated that endothelial cells contacted biocompatible bacteriophages which are found to migrate and spout into the ECM at the tube like structures. The function of angiogenesis was confirmed by angiogenesis factor of CD31 and VE-cadherin staining images. Our results suggested that biocompatible bacteriophage and ECM might continuously contribute to stimulate microenvironment for in vitro angiogenesis models. Also, we described that the functionalized bacteriophages can be used for feasible biomaterials on biomedical engineering fields. In the future, these studies are potentially applied for angiogenic matrix at the tissue engineering in vitro assay.

 

 

Speaker
Biography:

Jing An graduated from Chinese Medical University in 1982 in Shenyang of China. She got her Master’s degree and PhD in Clinical Medicine in 1989 at the Third Military Medical University in Chongqing (China), followed by a Postdoctoral position and Research Fellow position at the Department of Microbiology and Immunology in Tokyo (Japan) at the Metropolitan Institute for Neuroscience. In autumn 2000, she returned to China and occupies since the position of a Principal Investigator and Head of the Department of Microbiology at the Capital Medical University of Beijing (China). Her research focuses on the interaction between mosquito-borne flaviviruses and host as well as prevention of dengue virus infection.

 

Abstract:

Recently, Zika virus (ZIKV) causes millions of infections which emerge as a new dangerous member of the genus of Flavivirus. Unlike other well-known flaviviruses, ZIKV can be transmitted sexually and infect testes in murine models. However, the exact susceptible cells are not entirely clear. To investigate these issues, we infected interferon α/β and γ receptors deficient AG6 mice with ZIKV and examined the outcomes of infection. Infected mice displayed signs of reproductive system disorder, altered androgen levels in serum, and high viral load in semen and testes. Seminiferous tubules showed atrophy, accompanied by positive staining of ZIKV antigens on Sertoli cells. Viral particles and vacuole changes were observed within Sertoli cells, whose susceptibility to ZIKV was further validated in vitro study using cell lines. The disruption of tight junctions within testis and altered sperm morphology were also observed in ZIKV infected mice. Our results therefore demonstrated that Sertoli cells are susceptible to ZIKV infection, which results in the disruption of tight junctions in testis and causes abnormal spermatogenesis in mice. These results also imply that long-term impact of ZIKV infection on human male reproductive system requires close monitoring.

Speaker
Biography:

A Slonska Zielonka, PhD is currently working as Post-doctoral Researcher at Warsaw University of Life Sciences in the Faculty of Veterinary Medicine, Department of Physiological Sciences. She is interested in the field of mechanisms of herpesviral infections in primary murine astrocytes.

 

Abstract:

Equine herpesvirus type 1 (EHV-1) is a member of the subfamily Alphaherpesvirinae of the family Herpesviridae. In its target host, it induces mild respiratory diseases, abortion, neonatal foal death, and neuropathogenic disorders. EHV-1 has a broad host range in vitro, allowing for study of the mechanisms of productive viral infection, including endocytic pathways or intracellular transport in various cell cultures. The productive infection of astrocytes has been described for HHV-1, HHV-5 and HHV-6; however, there were no data about the ability of EHV-1 to infect the astrocytes. Recently, we reported for the first time that primary murine astrocytes were permissive to EHV-1 infection. Similarly to HHV-1, EHV-1 productively infected astrocytes and displayed cytopathic effect that resulted in the death of a portion of cell population. In the current study we investigated the mechanisms by which EHV-1 enters primary murine astrocytes. Using drugs that inhibit clathrin-dependent (chlorpromazine) or caveola-dependent endocytosis (nystatin), we showed that EHV-1 entry into murine astrocytes require clathrin, but not caveolae. The treatment of the cells with nystatin did not affect the replication of EHV-1. However, the use of chlorpromazine caused a significant decrease in the level of replication of EHV-1 detected by real-time PCR. In conclusion, EHV-1 efficiently entered and replicated in primary murine astrocytes. According to our results, we can assume that the principal pathway of EHV-1 entry into astrocytes appears to be caveolin-dependent and clathrin-independent.

 

Biography:

Sahar Essa has her experience and passion in improving the knowledge about respiratory viruses and viral immunopathology. Her published work reflects years of practice, experience and dedication in research. Her research shed the light on the impact of the circulating respiratory viruses and the cellular immune responses to Cytomegalovirus and Hepatitis C virus.

 

Abstract:

Hepatitis C virus (HCV) infection is a major public health problem with an estimated 3-4 million people infected each year worldwide. 20–30% of individuals acutely infected with HCV will spontaneously clear the virus, with the remaining 70–80% developing persistent HCV infection. The interplay between the virus and host innate and adaptive immune responses determine the outcome of HCV infection (Rauch et al., 2009). The present study aims to determine the level of cellular immune subsets in responders and non-responders HCV-infected patients as a result of the standard treatment (PEG-IFN and ribavirin) and to correlate the results with the major HCV genotypes in Kuwait. Data of the immunophenotyping for cellular subsets include 30 healthy controls and

 

genotype-4 HCV-infected patients (39 responders vs. 21 non-responders) at baseline and after treatment. The immunophenotyping was evaluated by flow cytometry using antibodies specific to mature T cells, T cytotoxic cells, regulatory T cells, T helper cells, activated T cells, natural killer cells, NKT cells and pan B cells. There were significant differences in the mean values of percentages for T helper cells, T cytotoxic cells, B cells, NK cells, NKT cells and activated T cells between HCV-responder vs. HCV-non-responder patients. Also, significant differences were noticed in the mean values of the absolute counts for T helper cells, B cells, NK cells, and T cells. Cellular subsets of the immune system play an important role in the pathogenesis, progression, and clearance of HCV. The screening for multiple cellular markers in the present study showed significant variations in the absolute counts and percentages of essential immune cellular subsets. These findings could lead to new possibilities for immune-based interventions and/or vaccine development with the aim of restoring functional antiviral T cell responses combined with improved viral clearance

Speaker
Biography:

M Chodkowski is a PhD candidate in the Division of the Veterinary Medicine in Warsaw University of Life Sciences. His main research topic is: Changes in the mitochondrial network in primary murine neurons infected with EHV-1. He is interested in molecular virology, neurovirology and novel treatment approaches using viruses e.g. oncolytic viruses.

 

Abstract:

Mitochondria have emerged as one of the key organelles in the maintenance of cellular homeostasis, innate immunity metabolism, aging, innate immunity, metabolism, apoptosis and other signaling pathway. In the last decade, work on the mitochondria has expanded our knowledge of its roles in cellular homeostasis in many parallel ways. There are not many report about interaction mitochondria – virus. Although all mitochondria have the same architecture, they vary greatly in shape and size. The mitochondria are composed of outer mitochondrial membrane, inner mitochondrial membrane, inter membrane space (space between outer and inner membrane), and matrix (space within inner mitochondrial membrane). The outer membrane is a smooth phospholipid bilayer, with different types of proteins imbedded in it. Mitochondria in eukaryotic cell formed network and they are distributed throughout the cells. In this study we examined changes of the mitochondrial network in HaCat cells during herpesvirus infection. For immunofluorescent staining, cells were plated onto laminin-coated coverslips. After 24, 48 h.p.i., infected cultures were fixed in 3.7% paraformaldehyde/PBS (Sigma Chemicals) for 30 min at room temperature. The cells were permeabilized in 0.5% Tween/PBS for 5 min, washed in PBS and blocked with PBS containing 1% bovine serum albumin (BSA) (Sigma Chemicals). Mitochondria were visualized using MitoRed (300 nM; Sigma Chemicals) and the cell nuclei were stained with Bisbenzimide /Hoechst 33258 according to the manufacturers recommendations. Results were evaluated under confocal microscope (Fluoview FV10i, Olympus). Due to infection with HHV-1 and HHV-2, the mitochondrial network is reorganized, both in the early and late stages of infection. Changes are manifested mainly through fragmentations of the mitochondrial network.

 

Speaker
Biography:

Golke A is an Associate Professor in the Department of Preclinical Science, Faculty of Veterinary Medicine, Warsaw University of Life Sciences. Her scientific interests include neuroinfections and the innate immune response to viral infections, in particular, the use of PRRs ligands in therapy of infectious diseases.

 

Abstract:

The body's ability to fight viral infection depends on the effective activation of the innate immune response. Activation of specific pattern recognition receptors (PRRs), including toll-like receptors (TLRs) triggers the production of pro-inflammatory cytokines, including interferons, which should lead to inhibition of viral replication. Because viruses have developed a number of mechanisms to interfere with TLR signaling, synthetic TLR activators can be considered as potential antiviral drugs. In the present study we have been investigating the effect of TLR synthetic ligands on the replication of equine herpesvirus type 1 (EHV-1) in ED cell line (equine dermal). Two different TLR ligands were used: TLR2/TLR6 ligand - Pam2CSK4 (0.1-100 μg/ml) and TLR9 ligand - ODN D-SL03 (1-10 μM/ml). The effect of TLR ligands on EHV-1 replication was assessed basing on the cytopathic effect and the quantitative evaluation of the number of viral DNA copies in the cells and the culture medium (real-time PCR). Both, on the basis of the cytopathic effect evaluation and the number of viral DNA copies quantification the antiviral activity of Pam2CSK4 at concentrations 10 μg/ml and 100 μg/ml was observed. No significant effect of ODN D-SL03 on EHV-1 replication was observed. However, preliminary observations based on the cytopathic effect evaluation showed that Pam2CSK4 at a concentration of 100 μg/ml in combination with ODN D-SL03 at a concentration of 10 μM/ml inhibited EHV-1 replication more efficiently than each of them individually. This stays with agreement with other studies, which show that it is necessary to stimulate both, TLR2 and TLR9 to trigger an effective immune response against herpes viral infections.

 

Speaker
Biography:

J Brzezicka is a PhD Student in the Department of Preclinical Science, Faculty of Veterinary Medicine, Warsaw University of Life Sciences. Her scientific interests include the impact of viral infection on the cytoskeleton, molecular virology and neuroinfections.

 

Abstract:

Adenoviruses are nonenveloped, double-stranded DNA viruses. Human adenoviruses (HAdV) are ubiquitous in populations worldwide. HAdV are classified into seven species (A to G). Due to the different tissue tropism, adenoviruses can be an etiological factor of infections of upper respiratory tract, digestive tract, urinary tract, eyes and the central nervous system. Children and adults with impaired immunity are particularly susceptible for infection. The aim of this study was to assess the changes in the actin cytoskeleton in A549 cells (adenocarcinomic human alveolar basal epithelial cells) after infection with different types of HAdVs. In the current study, three types of HAdVs were used: HAdV4, HAdV5 and HAdV7. Filament structures of actin were visualized using TRITC-phalloidin conjugate. Polyclonal antiserum ADENO MAB conjugated to FITC was used to detect viral antigens. Cell nuclei were stained with Hoechst 33258. Infected cells exhibited morphological changes, followed by cell lysis at the final step of infection. In A549 cells infected with HAdV4, 5 and 7 (at 12, 24 and 48 h p.i.), CPE consisted of disintegration and degradation of a nucleus, changes in a cell shape and rearrangements of the actin filaments. Furthermore, HAdVs used in this study caused actin accumulation in the nuclei of infected cells. Cells which did not undergo lysis showed high amounts of viral antigens in the cytoplasm. In the present study, we demonstrate that all used types of HAdVs are able to infect A549 cells, without the need for initial adaptation. The infection causes changes in cell morphology and cytoskeleton rearrangements. That may indicate that actin cytoskeleton is crucial for penetration into the cells, viral transport and transcription of viral genome.

 

 

Speaker
Biography:

J Cymerys, PhD is currently working as Associate Professor at Warsaw University of Life Sciences in the Faculty of Veterinary Medicine, Division of Microbiology. She is interested in the field of Viral Neuroinfections and Neurodegeneration.

 

 

Abstract:

Equine herpesvirus type 1 (EHV-1) is a main cause of respiratory disease, abortion and myeloencephalopathy in horses. Similarly to other alphaherpesviruses EHV-1 is neurotropic and causes latent infection in the neurons of natural host. Despite the fact that many studies have been devoted to the pathogenesis of various clinical forms of EHV-1 infection, mechanisms of the neuronal damage are not fully understood. In the present work, we examined one of the aspects of these disorders- tau-mediated neurodegeneration as a consequence of its hyperphosphorylation and sequestration in aggregates in infected primary murine neurons. Previous reports mainly focus on those HHV-1 infections which proved that tau protein (a microtubule-associated protein) in important in the neurodegenerative process. It is expressed in central nervous system neurons, where it plays a powerful role in regulating the dynamics of microtubule polymerization. Furthermore, it takes part in regulating axonal diameter, in axonal transport and in neurogenesis. When infected with HHV-1, tau protein may undergo modification, mainly via phosphorylation. As a result, phosphorylated tau protein isoforms aggregate and form neurofibrillary tangles, typical of neurodegenerative diseases called tauopathies. We describe, for the first time, that EHV-1 is capable of intensely modifying in the phosphorylation state of tau and that infection with EHV-1 leads to the accumulation of phosphorylated tau in the cytoplasm of neurons, especially during late hours after infection. This accumulation is dependent on the type of tau phosphorylation and on the time of infection.

 

 

Yu-Ju Lin

Centers for Disease Control, Taiwan

Title: National pandemic influenza preparedness plans in Taiwan
Biography:

Yu-Ju Lin is the Section Chief of the Division of Preparedness and Emerging Infectious Diseases, Centers for Disease Control in Taiwan (TCDC). She currently leads TCDC’s influenza pandemic preparedness and response activities, and her office is responsible for national and local readiness, the regulation of emergency operations, and managing the nation’s stockpile of related emergency countermeasures. She has more than 15 years of continuous service as a TCDC Officer; during her long career, she has taken part in a wide range of infectious disease outbreak investigations and responses, including SARS outbreak of 2003 in Taiwan, strategic development for the control of seasonal influenza/novel influenza A virus infections.

 

Abstract:

Background: The threat of a human influenza pandemic has prompted, thus, urgent development of national preparedness plans in Taiwan since 2003. We reviewed these plans to assess Taiwan’s preparedness for pandemic influenza.

 

Methods: The ‘National Influenza Pandemic Preparedness Plan’ and the ‘Strategic Plan’ of Taiwan, published in 2015 and 2011 respectively, were evaluated by using a checklist containing five criteria of preparation, surveillance, prevention and containment, case investigation and treatment, and risk communication (there are 68 indicators in total). This checklist was developed using the latest WHO guidelines, and in consultation with influenza experts of the WHO Eastern Mediterranean Office.

 

Results: The average score for aggregate completeness is 72.5%. For the five included criteria, the on average scores are 83.3% for preparation, 83.3% for surveillance, 57.4% prevention and containment, 71.4% case investigation and treatment, and 58.3% risk communication. Among the individual indicators, 31 (45.6%) indicators scored 3, 20 (29.4%) indicators scored 2, 15 (22.1%) indicators scored 1, and 2 (2.9%) indicators scored 0. Both of the 2 indicators scored 0 due to the lack of mention triage system and strategy for storage and disposal of corpses.

 

Conclusion: Taiwan’s preparedness plans are satisfactory, the plan scores particularly well on surveillance system and mobilization of resources such as health care facilities, personal protective equipment, antivirals, and vaccines. Moreover, gaps in preparedness planning remain; especially those operational guidelines for the implementation of related prevention and containment measures should be detailed or addressed in the operational documents.

 

Speaker
Biography:

Lamyaa Al-Dalawi has completed her Graduation in College of Veterinary Medicine, at University of Baghdad, Iraq. She worked as a Veterinarian in the Surgery Department of the Veterinary Medicine, University of Baghdad, Iraq. She has completed her Master degree in Internal and Preventive Medicine in the Veterinary College at the University of Baghdad. She later moved to the Medical Institute as a Lecturer in University of Kirkuk, Iraq. She obtained a scholarship to study PhD at Nottingham University. In October 2014, she started her PhD in the Department of Infection and Immunity/School of Veterinary Medicine and Science at the University of Nottingham. 

 

Abstract:

The influenza virus infection is influenced by a number of host cell factors, including host cell lipids. These lipids make up the bilayer membranes for both virus particles and host cells. The objective of this study is to determine the biophysical importance of lipids in terms of infectivity by pre-treating Influenza A viruses; avian influenza H2N3 virus, equine Influenza H3N8 virus and pandemic influenza H1N1 virus with various types of phospholipids. 1, 2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) had no significant impact on virus infectivity. However, 1, 2-dipalmitoyl-Sn-glycero-3-phospho-(1'-rac-glycerol) (DPPG) had a significant impact on H2N3, H3N8 and H1N1 infectivity. Treating the influenza viruses with lyso-analogues: 1-palmitoyl-2-hydroxy-sn- glycero-3-phospho-(1'-rac-glycerol) (LPG) and 1-palmitoyl-2-hydroxy-sn-glycero-3-phosphocholine (LPC) produced significant inhibition of influenza virus infection using MDCK cells and A549, that was dose-dependent. TEM images showed H2N3 and H3N8 without lipid pre-treatment are mostly spherical or filamentous, respectively. Incubating these viruses with lipids impact their morphology. Investigations of avian influenza H2N3 binding assay by flow cytometry demonstrates a high impact of the negatively charged phospholipids; i.e. either DPPG or LPG, blocking virus binding to cells significantly. Moreover, incubating influenza viruses with negatively charged phospholipids reduce cytokines expression especially IL-8. Overall, pre-incubating the virus with phospholipids seems to have an impact on the ability of the virus to bind to cells. So, specific lipids can be considered as a potential new inhibiting factor for influenza.