Day 1 :
University of New South Wales, Australia
Keynote: Improved molecular surveillance and new therapeutic responses to the influenza virus using mass spectrometry
Time : 10:05-10:40
Kevin Downard has obtained his Postdoctoral studies and held a subsequent academic position at the Massachusetts Institute of Technology after completing his PhD degree from the University of Adelaide, Australia. For the past 18 years he has held Professorial academic positions in the USA and Australia. He has over 100 publications including two books and is internationally recognized in his field.
The influenza virus is one of the deadliest pathogens known to man, responsible for the death of the equivalent of 1 in 1000 humans who have ever lived. Seasonal influenza accounts for about 3 to 5 million cases of severe illness requiring hospitalization and 250,000 to 500,000 deaths worldwide each year. A worldwide surveillance network, overseen by the WHO, assesses circulating strains and makes recommendations for the annual vaccine formulation ahead of the flu season in both the northern and southern hemisphere. Yet unforeseen evolutionary events, and growing resistance to current antiviral inhibitors, can lead the population unprotected. Furthermore, limitations in current screening technologies can delay and negatively impact on the implementation of effective infection controls. New molecular based surveillance technologies employing advanced mass spectrometry and bioinformatic approaches offer advantages for the characterization of circulating strains, the study of viral evolution and the identification and development of new antiviral inhibitors, including those based on natural products. This presentation will review these approaches that have attracted interest from global surveillance laboratories and have broader application to the study of other biopathogens which threaten human health.
The Scripps Research Institute, USA
Keynote: Broad neutralization of influenza viruses and progress towards a universal vaccine and therapy
Time : 10:55 - 11:30
Ian Wilson received a B.Sc. in Biochemistry from Edinburgh University, D. Phil. in Molecular Biophysics from Oxford University, and did postdoctoral research at Harvard University. Dr. Wilson has been a Professor at The Scripps Research Institute since 1982 and is Hansen Prof. of Structural Biology and Chair of the Dept. Integrative Structural and Computational Biology. His laboratory focuses on recogntion of microbial pathogens by the immune system and structure-based design of vaccines and therapeutics. Dr. Wilson is a Fellow of the Royal Society, Fellow of the Royal Society of Edinburgh, Member of the American Academy of Arts and Sciences, has a D.Sc. from Oxford University, and published over 665 papers.
The major surface antigen, the hemagglutinin (HA), of influenza virus is the main target of neutralizing antibodies. However, until recently, most antibodies were thought to be strain-specific and protect only against highly related strains within the same subtype. However, in the past few years, many human antibodies have been isolated that are much broader and neutralize across subtypes and groups of influenza A and B viruses through binding to functionally conserved sites. We have determined structures of many broadly neutralizing antibodies with HAs and determined that their epitopes map to highly conserved sites on the HA fusion domain (stem) and receptor binding site (head). The identification and characterization of the epitopes and mode of binding of these antibodies have elucidated recognition motifs and conserved sites of vulnerability that provide exciting new opportunities for structure-assisted vaccine design as well as for design of therapeutics that afford greater protection against influenza viruses.
Nanorx Inc, USA
Time : 11:30-12:05
Palayakotai Raghavan is CEO and Founder of Nanorx INC. He completed Ph.D in Organic Chemistry from Oregon State University (1979) and M.S in Chemistry (1972) from I.I.T Mumbai, India. He has worked on drug discovery for over 25 years at Columbia University, Max-Planck Institute, Germany, Ciba-Geigy (now Novartis) and Boehringer Ingelheim. He has over 12 patents and another 15 pending patent applications.
Metadichol (US patent 8,722,093) is a Nano emulsion of long-chain alcohols found in many foods. It is commonly called Policosanol and is present in foods such as rice, sugar cane, wheat, peanuts Metadichol acts on Nuclear Vitamin D receptors (VDR) (US patent 9,006,292) that are present in cells throughout the body to stimulate the immune system and inhibit a variety of disease processes, resulting from viral infections.We tested for antiviral activity of Metadichol® in Vero and MDCK cells infected with Influenza A, H1N1, Human Respiratory Syncytial viruse, Dengue, Chikungunya and. Ebola, Marburg. In addition, we tested the efficacy of Metadichol® in preventing cell death caused by Adenovirus, Tacaribe Mammarena virus, Rift Valley Fever virus, SARS coronavirus, Japanese Encephalitis virus, West Nile virus, and Yellow Fever virus. In the in vitro assays, Metadichol showed no cytotoxicity and strongly inhibited cell death caused by each of the viruses tested. Metadichol is a safe and effective inhibitor of enveloped viruses in humans. Since it is known to bind to the vitamin D receptor (VDR) (US patent 9,006,292), its mechanism of action likely involves the competitive displacement of virus particles from VDR’s on host cell membranes. Because it consists of natural components of common foods and has no known negative side effects, Metadichol has the potential to serve as a novel, broad-spectrum antiviral treatment for Dengue, Ebola, Zika, H1N1, SARS, Chikungunya and other enveloped viruses.
- Track 1: Influenza Vaccines: Designs and Developments Track 3: Influenza: Causes, Symptoms and Treatment Track 5:Influenza Vaccines : Safety and Effectiveness Track 7:Advances in Viral Detection and Identification Technologies Track 9:Host Genetics of Infection and Immunology
Nanorx Inc, USA
Institute of Animal Reproduction and Food Research of Polish Academy of Science, Poland
Cilian AG, Germany
Title: CiFlu®: Development of a novel subunit influenza vaccine candidate based on the ciliate performance expression system
Time : 12:05-12:30
Marcus Hartmann has spent his scientific career investigating protozoan organisms, particularly Ciliates. He worked for the central research department of Aventis, Frankfurt (Germany) and was a postdoc in an academic working group at the University of Münster. His postdoctoral study dealt with research in the field of commercial applications of protozoan organisms. Based on his extensive scientific experience in the field Ciliate biotechnology, he founded Cilian, and since then he has headed the Company’s R&D team. Marcus Hartmann is the author of numerous scientific publications, recitations and patents in the area of Ciliate biotechnology. One of the main breakthrough of his team was the first-ever production of therapeutically usable proteins in Ciliates.
The critical annual manufacturing process for seasonal influenza vaccine based on embryonated chicken eggs, involves numerous steps and takes on average 6 to 8 months to complete. This often means that vaccine is only available late into the flu season. The timely availability of an effective influenza vaccine, at or before the flu season starts, is even more acute for vulnerable highest risk groups such as persons 65 years of age and older. The lack of timely availability of seasonal/pandemic vaccine has raised significant questions about the utility of the current, antiquated, cumbersome, expensive and unsafe manufacturing platform involving chicken eggs. Safety concerns about cell culture based virus proliferation processes called also alternative flu vaccines production processes into question. Now new recombinant antigen manufacturing platforms were postulated to reduce production time and costs. Cilian’s flu vaccine CiFlu® is a cost-effective subunit vaccine based on the heterologous expression of recombinant Influenza hemagglutinin (rHA) in the ciliate Tetrahymena. Utilizing its CIPEX-System as such a manufacturing platform, Cilian has successfully demonstrated repeated expression of rHA at high yield: four subunit vaccines has been expressed and shown to be functionally active. Mice were first immunized with the monovalent rHA. HA antibodies were harvested and its ability to inhibit the respective influenza strain was tested. The results demonstrated comparable or better efficacy (in vivo inhibitory immunogenicity) to monovalent vaccine from chicken eggs. Cilian meanwhile received a positive scientific advice from the German Paul Ehrlich Institute for CiFlu® and is developing a comprehensive clinical plan.
University of Chicago Medicine, USA
Time : 12:30-12:55
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.
Clinical recognition of severe respiratory illness (SRI) is difficult. Influenza-like respiratory illness often masquerades as asthma, especially in patients with and without pre-morbid pulmonary disease. Because the initial differential diagnosis includes asthma, this can lead to treatment confusion and an underestimation for the primary cause for SRI. Viral bronchosplasm is difficult to ameliorate air-flow obstruction (AFO) with bronchodilator therapy that is refractory to beta-agonist and steroid therapy. This may lead to patients requiring supportive respiratory care. Viral SRI is now documented to come from multiple viral sources and appear in different packages. These viruses are high pathogenic and attack the bronchial wall structure causing hypercarbic respiratory failure. These viral infections are now documented as being the etiology of global epidemics and pandemics. In August 2014, there was an increase in SRI that was associated with Enterovirus EV-68 which was reported in 41 states across the United States. The Center for Disease and Control (CDC) received over 2600 samples, 36% positive for EV-68. Viral illnesses have a huge impact on global resources and finances. Clinical diagnoses via respiratory viral panel and chest radiography may be relevant. Newer treatment plans for SRI include the use of high flow nasal cannula (HFNC) and heliox. Failure to recognize SRI may lead to hypercarbic respiratory failure where the support therapy is ventilator, pruning, nitric oxide, ECMO. This can lead to complications such as; ARDS, kidney and other organ failure and severe acute respiratory syndrome. More study is needed to understand the relationship between SRI-AFO.
Polish Academy of Science, Poland
Time : 12:55-13:20
Hanna Radecka was graduated from the Department of Chemistry of Nicolaus Copernicus University in Toruń in 1978. She was a Visiting Scientist at the Hokkaido University in Sapporo and at the University of Tokyo. Since 1998, she is working at Department of Biosensors of the Polish Academy of Sciences in Olsztyn. In 2011 she received the title of Professor of Analytical Chemistry and was nominated as the Head of Laboratory of Bioelectroanalysis. Currently she is working on the development of the new biosensors for determination of avian influenza viruses, possible biomarkers of Alzheimer’s and other neurodegenerative diseases present in human plasma.
Here, we report examples of successful developing of several type of immunosensors destined for the detection of Highly-Pathogenic Avian Influenza type H5N1 virus (HPAI) spreading among wild and domestic birds. The immunosensor were developed by the successive modification of gold as well as glassy carbon electrodes. The whole antibody or their fragments have been applied as the sensing elements. The complex between virions and specific antibody adsorbing on a surface of an electrode forms an insulating layer. This phenomenon, which is a base of ion-channel mimetic type of immunosensors, can be monitored by the electrochemical impedance spectroscopy (EIS) in the presence of [Fe(CN)6]3-/4- as a redox marker. The another type of immunosensors are based on redox active layers incorporated di-pyrromethene-Cu(II). The changes of electrochemical parameters of redox centers upon target analyte binging are the base of analytical signal generation. The both type of immunosensors displayed better sensitivity towards viruses as well as antibodies in comparison to ELISA; they are also very selective. The matrix from hen sera has no influence on the immunosensors performance. In addition, very small analyzed sample volumes (10 µl) are needed. After miniaturization, they keep excellent analytical parameters. Therefore, immunosensors presented could be recommended for the direct electrochemical detection of viruses as well as antibodies in the natural physiological samples.
Polish Academy of Science, Poland
Title: Electrochemical genosensors based on redox active monolayers: Characterization and applications
Time : 14:05-14:30
Jerzy Radecki is the Professor of Analytical Chemistry and currently working as Head of Department of Biosensors of IARFR PAS in Olsztyn. His research interest concerns the developing of new sensors and biosensors based on the intermolecular recognition processes occurring at the border of the aqueous and organic phase. Particularly, he is interested in functionalization of surface of solid electrodes with “host” molecules, which are responsible for “guest” molecules (analytes) recognitions. He is working on not only analytical aspects of developed sensors but on the elaboration of the mechanism of analytical signal generation as well.
Here we report on electrochemical genosensors devoted for detection of influenza virus H5N1 gene sequence. Using ssDNA decorated with redox active units such as Co(II)-porphyrin or 3-iron bis (dicarbiollide), the detection limit in the fM range has been achieved. The strategies based on dipyrromenthene Cu(II) redox active monolayer or phenanthroline-Epoxy-Fe(III) complexes have been also applied for the development genosensors destined for detection of DNA as well for RNA derived from Avian Influenza viruses. They have been working based on the new “ion barrier switch-off” mechanism of analytical signal generation. To face of the need of systems for simultaneous determination of few markers of one disease coming from medical diagnosis, we have developed a novel dual DNA electrochemical sensor with “signal-off” and “signal-on” architecture for simultaneous detection of two different sequences of DNA derived from Avian Influenza Virus type H5N1 by means of one electrode. Two sequences of ssDNA characteristic for hemagglutinin decorated with ferrocene and characteristic for neuraminidase decorated with methylene blue were immobilized covalently together on the surface of one gold electrode. Taking into account the excellent analytical parameters of genosensors presented such as good sensitivity, selectivity and very low sample consumption, they could be recommended for future wide application for medical diagnostic as well as environmental control.
The University of Adelaide, Australia
Time : 14:30-14:55
Farhid Hemmatzadeh joined The University of Adelaide as a senior lecturer of virology at the School of Animal & Veterinary Sciences in 2009. Previously, he was employed by Melbourne University since 2005 and Tehran universities as an associate professor since 1997. He has over 20 years experience in research and teaching at the field of animal viral diseases including herpesviruses, pestiviruses, retroviruses, parvoviruses and influenza viruses. Farhid has been involved in research, development and assessment of diagnostic test for animal viral diseases specially DIVA tests for poultry and large animals.
In last 15 years numbers of ELISA test were developed to differentiate influenza infected from vaccinated animals (DIVA). In most of the test the either viral associated infection antigens or heterologous Neuraminidase antigens were used to develop DIVA tests. One of the first attempts was non-structural 1 protein (NS1). The NS1-based ELISA was shown reliable results as a DIVA test in young chickens but the accuracy of NS1-based DIVA test decreases by the time and numbers of vaccination produces non-specific reactions. Nucleoprotein (NP) and conserved HA274–288 epitope were the others candidates for DIVA test but these two antigens didn’t show any values as DIVA ELISAs. By now the best antigen to develop DIVA-ELISA test is ectodomain of matrix 2 (M2e) protein. Relatively invariable nature of M2e protein across AIV strains and high level of expression of M2e protein on the surface of infected cells despite being low in copy number in a mature virions are the main properties that make M2e a suitable candidate for DIVA tests. Our studies on structure of M2e showed the tetramer form of M2e shows higher sensitivity and specificity to discriminate M2e antibodies in sera of infected birds from vaccinated or non-vaccinated birds.
The Ragon Institute of MGH, MIT and Harvard University, USA
Title: Multivalent influenza hemagglutinin promotes the immundominance of non-neutralizing antibody responses through reptatively constrained orientation
Time : 14:55 - 15:20
Daniel Lingwood is an Assistant Professor at The Ragon Institute of MGH, MIT and Harvard and is a faculty member in the Virology Program at Harvard Medical School. He received his Ph.D. from the Max Planck Institute for Molecular Biology and Genetics, and conducted postdoctoral work at the Vaccine Research Center at NIH. Dr. Lingwood has garnered international recognition for his discovery that humans possess genetically-encoded antibody sequences that when properly oriented as germline B cell receptors, naturally engage conserved sites of viral vulnerability and serve as substrates upon which broadly neutralizing antibodies can be developed.
Much of the influenza virion surface is occupied by a dense array of trimeric hemagglutinin (HA) that functions to engage sialyl-oligosaccharide on a target cell. This dense packing of spike protein is also thought to restrict antibody access to the conserved HA stem epitope, a weakly immunogenic target for broadly neutralizing antibody (bnAb) responses against this virus. However, recent cryo-EM studies, have suggested that stem-directed bnAbs do not have restricted access to this site. To functionally define the source of weakened immunogenicity to the stem epitope, we compared stem specific antibody responses to three structurally-defined presentations of HA: soluble trimer, and ferritin nanoparticle 8mers displaying either the full-length trimer or stem/stalk region alone. Surprisingly, we found that while the nanoparticles were more immunogenic, only the soluble trimeric format elicited detectable stem-epitope directed antibodies upon initial exposure to antigen. We propose that antigen multivalency, a cornerstone of both vaccine design and viral architecture, imposes not only repetitive array to increase immunogenicity but also restricted antigen orientation, which can limit exploration of antigenic space, insuring that immunodominant non-neutralizing responses are non-linearly amplified during this process. Repetitive exposure to the soluble HA trimer eliminates reactivity to stem due to amplification of immunodominant non-stem responses; our work shows that multivalent HA display can achieves the same result within a single encounter. These data highlight a previously unrecognized mode of immune distraction and delineate the relationship between antigen valency and the target-specificity of the humoral response.
Mechnikov Research Institute of Vaccines & Sera, Russia
Time : 15:20-15:45
Yuri Vasiliev has completed his PhD from the Moscow Medical University in prepandemic avian influenza vaccines at the age of 24 years and started postdoctoral research at the Mechnikov Research Institute of Vaccines and Sera, Moscow. He became head of laboratory of experimental immunology at the age of 28 years and a year later was appointed as head of R&D for adjuvants, his current position at the Mechnikov Institute. He has published more than 50 works locally and internationally, with the top citations per single paper reaching 30 (bibliometrics via Web of Knowledge).
Chitosan-based formulations combine effectiveness, safety and economic feasibility, and have been studied as vaccine adjuvants and gene delivery systems. However, chitosan is an umbrella term for a very diverse group of glucosamine-based substances and their derivatives as well as adjuvants (e.g. solutions, particles). Lack of consensus on nomenclature and standardization approaches renders juxtaposition and reproduction of data across various studies nearly impossible, and underlying mechanisms of action for chitosan-based adjuvants remain largely unknown. Panels of chitosan substances and chitosan-based adjuvants (currently exceeding 50) have been created and characterized extensively using distinct methods (HPLC, NMR, etc.). Datasets generated from large-scale preclinical studies in various animal models demonstrate that principal chitosan characteristics (molecular weight and deacetylation degree) determine adjuvant properties through a very complex interaction. No single characteristic is responsible for high or low immunogenicity, effectiveness (lethal challenge model) and safety. Levels of serum and lung antibodies, IgG subclasses and certain cytokines and, thus, Th polarization also varied for different chitosans. Certain chitosans and derivatives (e.g., succinylated) were not immunogenic at all. Impurities (e.g., endotoxins, proteins) were challenging to evaluate due to interference from chitosan, however, did not have a critical effect on adjuvant properties. A universal chitosan-based adjuvant has been developed and successfully evaluated with various vaccines against influenza (subunit, cold-adapted, etc.) and other human and animal infections as well as in comparison with other adjuvants (aluminium-based, oil-in-water emulsions, etc.). Chitosan-based adjuvants tailored for certain types of influenza vaccines (100-fold increase of immunogenicity) and complex formulations are also being studied.