Day 2 :
Keynote Forum
Walter Doerfler
University of Erlangen-Nuernberg & Cologne University, Germany
Keynote: Discoveries with the adenovirus type-12 system: Integration of viral DNA and epigenetic consequences
Time : 10:00-10:30
Biography:
Walter Doerfler has completed Munich Medical School (LMU), Doctor of Medicine thesis in Anatomy and Internships in Munich and Trenton, NJ, USA. He did his Postdoctoral studies in Molecular Genetics at the Max-Planck-Institute for Biochemistry in Munich and in the Department of Biochemistry, Stanford University Medical School.
Abstract:
The human adenovirus type-12 (Ad12) system has been used to investigate basic mechanisms in molecular biology and viral oncology. Ad12 replicates in human cells, but undergoes a completely abortive cycle in Syrian hamster cells. Ad12 induces neuro ectodermal tumors in newborn hamsters (Mesocricetus auratus). Each tumor cell or Ad12-transformed hamster cell carries multiple copies of integrated Ad12 DNA. Ad12 DNA usually integrates at one chromosomal site which is not specific; Ad12 DNA can integrate at many different locations. Virion or free intracellular Ad12 DNA remains unmethylated at CpG sites, whereas the integrated viral genomes become de novo methylated. Inverse correlations between Ad12 promoter methylation and activity were described for the first time in this system and initiated active research in the field of DNA methylation and epigenetics. Promoter methylation is an important factor in long-term gene silencing. We have also discovered that the insertion of foreign (Ad12, bacteriophage lambda, plasmid) DNA into mammalian genomes can lead to genome-wide alterations in methylation and transcription patterns in the recipient genomes. This concept has been verified recently in a pilot study with human cells which had been rendered transgenomic for a 5.6 kbp bacterial plasmid. Currently, we study epigenetic effects in Ad12 infected cells and in Ad12 induced hamster tumor cells. These genome-wide epigenetic alterations are considered crucial elements in (viral) oncogenesis.
Keynote Forum
Angelika Krumina
Riga Stradins University, Latvia
Keynote: Molecular Genetic Characteristics Patients with Borrelia burgdorferi Infection
Time : 10:30-11:00
Biography:
Angelika Krumina is a leading Researcher in Infectology field in Riga East Clinical University Hospital. She is a Scientific Expert of Latvian Academy of Sciences, Board Member of Latvian Medical Association and Member of European Society of Clinical Microbiology and Infectious Diseases. During 2011-2012, she was a Project Coordinator in State Program of Latvian Academy of Sciences. Currently she is a Work Package 6 Leader in Latvia of Baltinfect project of the Seventh Framework Program (FP7) of the European Commission.
Abstract:
The MHC II genes encode the polymorphic-DR and DQ molecules, Lyme disease (Lyme borreliosis) is infectious vector-borne diseases which has a large polymorphism of clinical manifestations and caused at least three species of bacteria of the genus Borrelia: Borrelia burgdorferi sensu stricto dominated as the causative agent of Lyme disease in the United States, whereas Borrelia afzelii and Borrelia garinii in Latvia and Europe. One of the major unsolved problems of today is the study of the interactions of organism and pathogens genospecies Borrelia burgdorferi (Bb). In this connection particular interest is the analysis of one of the major systems of the body controlled by the immune response-complex HLA including the identification of possible associations of HLA genotypes with clinical features of Lyme disease. Although various factors participate in the immune response to infection (e.g., intensity of the infection and T-cell functioning), the interaction of these mechanisms with genetic factors seems to be important in determining the evolution of the disease. Furthermore, the immune response to infections varies from one individual to another, on account of the polymorphism of the genes that influence this response. HLA antigens may act alone or in combination with other genes, conferring susceptibility to, or protection against infectious diseases. The obtained results suggest that the inflammatory events of the sub acute arthritis can set the stage for development of chronic disease in individuals possessing an risk haplotypes. In particular, the haplotypes HLA-DRB1*15:01:01/DQA1*01:02:01/DQB1*03:02:01, (OR=8.34; p<0.013); -DRB1*01:01:01/DQA1*03:01:01/DQB1*03:02:01, (OR=6.17; p<0.027) and -DRB1*03:01:01/DQA1*01:01:01/DQB1*05:01:01, (OR=2.66; p<0.032) contributes definitely to a genetic predisposition to Borrelia burgdorferi infection in Latvian population which may have implications in our understanding of pathogenesis of this disease. Knowledge of the mechanisms of genetic protection against and susceptibility to infectious diseases is one of the important steps towards controlling them in endemic areas and contributes to our understanding of both the pathogenic and protective mechanisms of these processes. The mechanisms of immune response to infection that are influenced by the HLA genes may be the key to future vaccines using the peptides of organisms that mimic the HLA antigens.
Keynote Forum
Bojian Zheng
The University of Hong Kong, Hong Kong
Keynote: A Novel Mutation D404N in the connection Sub-Domain of Reverse Transcriptase of HIV-1 CRF08_BC subtype Confers Cross Resistance to NNRTIs
Time : 10:40-11:10
Biography:
Bojian Zheng has completed his PhD at The University of Hong Kong and Postdoctoral studies from McMaster University in Canada. He is currently a Professor working in Department of Microbiology, The University of Hong Kong. He has published more than 180 papers in reputed journals and has been serving as an Editorial Board Member of several international journals.
Abstract:
Growing evidence suggests that mutations in the connection domain of the HIV-1 reverse transcriptase (RT) can contribute to viral resistance to RT inhibitors. This work is designed to characterize a novel mutation D404N in the connection sub-domain of RT of HIV-1 CRF08_BC subtype on drug resistance, viral replication capacity (RC) and RT activity. Mutation D404N alone or together with the other reported mutations were introduced into an HIV-1 CRF08_BC subtype infectious clone by site-directed mutagenesis. The drug susceptibility to nine RT inhibitors, viral RC and the DNA polymerase activity of viral RT of the constructed virus mutants were investigated. Modeling study using the server SWISS-MODEL was conferred to speculate the possible structure-related drug resistance mechanism of the mutation D404N. Single mutation D404N and H221Y conferred low-level resistance to nevirapine, efavirenz, rilpivirine and zidovudine. Double mutations Y181C/D404N and Y181C/H221Y significantly reduced susceptibility to NNRTIs. The most pronounced resistance to NNRTIs was observed with the triple mutations Y181C/D404N/H221Y. The virus containing a single mutation D404N displayed approximately 50% of RC compared to the wild type (WT) virus. Modeling study suggested that the D404N mutation might abolish the hydrogen bonds between residue 404 and K30 in p51 or K431 in p66, leading to impaired RT subunit structure and enhanced drug resistance. These results indicate D404N to be a novel NNRTI-associated mutation in the HIV-1 CRF08_BC subtype which provides valuable information to monitor clinical RTI resistance.
Keynote Forum
Kunihiro Kaihatsu
Osaka University, Japan
Keynote: Sequence Specific Detection Of Rna Viral Gene By Chemically Modified Peptide Nucleic Acid
Time : 11:00-11:30
Biography:
Kunihiro Kaihatsu has received his PhD in Organic Chemistry and Nucleic Acid Chemistry from Kobe University, Japan. He has served as an Associate Professor in the Department of Organic Fine Chemistry in the Institute of Scientific and Industrial Research, Osaka University, Japan. He has received several awards from Chemical Society of Japan, International Society of Antiviral Research, Biobusiness Competition Japan Award in 2009 & 2010 and other research societies. He is currently focusing on developing diagnostic methods in virology using nucleic acid chemistry.
Abstract:
Molecular tools that can rapidly identify virus in clinical specimen are important for achieving therapeutic success and preventing the spread of infectious diseases. Peptide nucleic acid (PNA) is a DNA/RNA analogue that possesses a neutral amide backbone instead of phosphate backbone and thus efficiently binds to DNA without having electrostatic repulsion. Currently, we synthesized PNAs derivatized with different types of intercalators via an amide linkage at the N-terminus. Their intercalators increased PNA binding affinity to matched DNA; however, most of them also increased the binding efficiency to mismatched DNA. On the other hand, PNA derivatives with tolane selectively improve the binding affinity to the matched sequence, but not to the mismatched sequence. In this paper, we synthesized a series of PNA-tolane conjugates and measured UV melting temperature (Tm) with target ssDNA and ssRNA. Molecular dynamics simulations and thermodynamic studies were also performed to discuss the mode of actions of PNA-tolane derivatives against ssDNA and ssRNA. PNA-tolanes were also used to discriminate a neuraminidase inhibitor-resistant influenza A virus gene on our novel type of nucleic acid chromatography system.
Poster Session 11:50-13:10
Lunch Break:13:10-13:55 @ Salamanca
- Therapeutic Approaches & Targets for Viral Infections
Veterinary Virology
Clinical & Neuro Virology
Agriculture and plant virology
Location: Madrid, Spain
Chair
Kunihiro Kaihatsu
Osaka University, Japan
Co-Chair
Doerfler Walter
University Erlangen-Nuernberg, Germany
Session Introduction
Beti Ernawati Dewi
Universitas Indonesia, Indonesia
Title: Development of animal model for dengue virus antiviral trial using infected cells
Time : 13:55-14:20
Biography:
Beti Ernawati Dewi has completed her PhD at the age of 37 years from University of Tsukuba, Japan and postdoctoral studies from Institute of Tropical Medicine, University of Nagasaki, Japan. She is a teaching staff and senior researcher at Department of Microbiology, Faculty of Medicine, Universitas Indonesia. She has published more than 10 papers in reputed journals.
Abstract:
Dengue virus (DENV) infection is still a serious health problem in Indonesia with no specific antiviral to treat the patients. Due to the lack of a good animal model, the development of antiviral has not been completely elucidated, In this study we develop a mice model of DENV infection for antiviral experiment. DDY mice were injected with 5.5 x 106 DENV-2 New Guinea C infected Huh-7 and Vero cell via intra peritoneal. The presence of DENV in plasma sera were evaluated by focus assay at 6, 24 and 48 hours post infection. In mice with DENV-2 infected Vero cells, the titer of DENV-2 was 1.7x102 FFU/ml at 6 hours post infection. At 24 and 48 post infection no DENV were found in plasma mice with DENV-2 infected Vero cells. The higher titer was found in mice with DENV-2 infected Huh-7 cells with 5.8 X 103FFU/ml and 3,6X103 FFU/ml at 6 hours and 24 hours, respectively. The DENV viremia results demonstrate that DDY mice injected with DENV infected Huh-7 are a promising simple animal model for DENV antiviral trials in future. Keywords: Animal Model, Dengue virus, Vero cell, Huh-7 cells
Sharad Kumar Yadav
U. P. Veterinary University, India
Title: Scenario of immunodiagnostics for animal herpes viruses
Time : 14:20-14:45
Biography:
Sharad Kumar Yadav has 25 years of teaching and research experience and has served to various senior positions of the University including Registrar of the Veterinary University, Mathura, India. He is currently the Director at Cow Research Institute and Professor and Head of Department of Veterinary Microbiology at Veterinary University, India. He has published number of papers in reputed international & national journals and has a vast experience in the arena of BHV-I virus research.
Abstract:
Bovine Herpesvirus Type-1 (BHV-1), aetiological agent of a number of diseases including IBR, infectious pustular vulvovaginitis (IPV), infectious balanoposthitis (IBP), conjunctivitis, encephalomyelitis, mastitis, abortion, enteritis and interdigital lesions of domestic and wild cattle. Knowledge of genomic sequence of the virus, virus-specified components and their concerted action are helpful in predicting and defining the chain of events during the lytic and latent phases of the BHV-1 and determining the strategies through which bovine Herpes viral life cycle can be interrupted. Investigating herpes viruses would surely reveal most useful targets for formulating ways of diagnosis, prevention and antiviral treatment against herpes viruses. Following infection, BoHV-1 may persist in infected animals in a latent state in sensory neurons, e.g., in the trigeminal or sacral ganglia. Rapid and precise diagnosis plays a pivotal role in implementing suitable control measures in natural field cases of bovine abortion due to infection with BHV-1. The diagnosis of BoHV-1 associated abortions is commonly made through the observation of gross and histopathological lesions in fetal and placental tissues, by the detection of viral antigen by fluorescent antibody test (FAT), immunohistochemistry (IHC), or viral DNA by nested PCR and real-time PCR, multiplex real-time PCR for differentiation of field and vaccine virus strain. After virus isolation virus can be detected in cell culture by virus isolation, serology, histopathology, immunohistochemistry (IHC) and real-time polymerase chain reaction (PCR). Other diagnostic applications include strain characterization by restriction enzyme fragment length polymorphism (RFLP) analysis, antibody detection by neutralization test and ELISA, differentiating antibody analyses by gB blocking ELISAs, indirect ELISAs and gE-blocking-ELISAs. To deepen the dialogue between the authorities, the health and medical/veterinary care system, the academic sphere and industry must act altogether by establishing a “Task Force†at authority level for well-timed diagnosis and henceforth prevention of herpes viral infections.
Raquel Rodrigues
Umeå University, Sweden
Title: Defining the nature of the cellular receptor for LGTV
Time : 14:45-15:10
Biography:
Raquel Rodrigues has completed her PhD at the age of 28 from the École Normale Supérieure de Lyon in France and is now undergoing postdoctoral studies at the Department of Clinical Microbiology of the Umeå University in Sweden. She has published several papers in virology journals and has presented her findings in virology and infectious diseases conferences.
Abstract:
The tick-borne encephalitis virus (TBEV) is the most important endemic arthropod-borne flavivirus in Europe. It provokes tick-borne encephalitis (TBE), an infection that can cause mild to severe forms of neurological diseases such as meningitis or encephalitis. However, the life-cycle of the virus and in particular the first steps of infection are still poorly understood. To better understand the life-cycle and pathogenesis of TBEV, identification of the virus-host interactions such as attachment to and entry into host cells are of utmost importance. In this study, we used Langat virus (LGTV), the naturally attenuated form of TBEV, as a model to investigate the nature of the cellular receptor(s) used by TBEV for attachment and entry. For that, we treated cells with compounds that cleave or inhibit several receptor types (proteins, N-glycans, O-glycans, glycolipids, sulfated glycans and sialic acids) and measured the attachment of LGTV. Also, we made use of CHO cells lacking N-glycans or sialic acids to quantify the binding of LGTV. According to our results, the cellular receptor for LGTV seems to include a protein component, as shown by the decrease in binding observed after treatment with proteases. Furthermore, no decrease in binding was found after inhibition of the glycans, which suggests that glycans are not involved in the attachment of LGTV to host cells. Our results strongly suggest that the receptor for LGTV is of protein nature. Based on these results, a proteomic approach is currently ongoing in order to identify the specific receptor protein for LGTV.
Amal Souiri
University Hassan II of Casablanca, Morocco
Title: Genetic characterization of Pepino mosaic virus infecting Moroccan tomatoes
Time : 15:10-15:35
Biography:
Amal Souiri is currently a PhD Student from the Faculty of Science, University Mohammed V in Morocco. She is a Young Researcher on Virology, Molecular Biology and Immunology. She has contributed in the development of monoclonal antibodies against Pepino mosaic virus infecting tomato crops in Morocco and the genetic characterization of Moroccan PepMV strains. She has published 3 papers in reputed journals and communicated its studies in many international congresses.
Abstract:
Over the past few years, Pepino mosaic virus (PepMV) has become an emerging pathogen that causes significant losses in tomato crops worldwide. Genetic composition of PepMV population in Morocco has not been determined yet. In this study, twelve PepMV isolated from Moroccan areas of tomato production were identified by serological and molecular tools and were characterized by nucleotide sequences analysis of a part of RNA-dependant RNA polymerase gene, triple gene block and coat protein gene. In spite of genetic diversity of PepMV isolates due to high mutation and recombination rate of RNA, the phylogenetic analysis based on comparison with the known genotypes showed that the Moroccan population of PepMV shares a very high sequence identity with CH2 strains. As well, Moroccan isolates reveals a specific single nucleotide polymorphisms that lead to distinct variants and for a subset of isolates, a possible recombination with EU genotypes. This study will contribute in the improvement of control strategies of PepMV disease and in the implementation of phytosanitary requirements for the exported tomato crops.