Day 1 :
Jaroslav Turánek has his expertise in nanotechnology focused on drug delivery systems based on liposomes for construction of self-assembled vaccination nanoparticles and molecular-based adjuvants. He is pioneering the technologies of nanofibre-based mucoadhesive films for noninvasive mucosal vaccination and “printed vaccine technology”. Jaroslav Turánek has completed his PhD at the age of 27 years from Masaryk University Brno. He is the head of Department of Pharmacology and Immunotherapy, Veterinary Research Institute, Brno. He has published more than 75 papers on vaccines, drug targeting, anticancer and antimicrobial drugs in reputed journals. He is teaching immunology, biotechnology and immunochemistry at Masaryk University Brno and Technical univeersity Brno. He is president of Czech Society for Gene and Cell Therapy and Principal Investigator of vast multidisciplinar project OPVVV FIT “Pharmacology, Immunotherapy, nanoToxicology” focused on applicaton of complex nanotechnology and biotechnology approach for development of new modern vaccines and targeted drug delivery systems.
Aim: The aim is to present the latest developments in the field of noninvasive mucosal vaccination. Selected speakers will cover topics like systems for mucosal vaccination, mucosal molecular adjuvants, mRNA constructs for vaccination, influenza virus pseudotypes, recombinant antigens and antigen formulations for mucosal delivery.
Statement of the Problem: Vaccination remains the principal way to control seasonal flue infections and is the most effective method of reducing influenza-associated morbidity and mortality. Mucosal vaccination represents non-invasive route for immunization having advantage in safety, efficacy and comfort for vaccinees, in general. At present, inactivated, live attenuated and recombinant vaccines for intradermal, intramuscular and intranasal application are licensed and in use.
Methodology & Theoretical Orientation: Advent of reverse vaccinology together with availability of recombinant technologies for economic massive production of recombinant antigens and mRNA constructs, new biocompatible nanomaterials and molecular adjuvants are prerequisites for successful development and commercialization of influenza vaccines. Especially mRNA-based influenza vaccines represent promising approach for rapid development of seasonal influenza vaccine. Recombinant antigens based on complex nanoparticles like VLP and virus pseudotypes, recombinant protein antigens derived from influenza virus and chimeric multiepitopic or fused complex protein antigens represent valuable tools for development of modern influenza vaccines. Beside intranasal vaccination, sublingual application of vaccines is of growing interest with respect to safety and induction of both systemic and generalized mucosal immune response. Development and application of new mucoadhesive films drives forward the development of mucosal vaccines. Great potential is hidden in the use of biocompatible nanomaterials, especially nanofibre- based cloth. Polymeric and lipid based nanoparticles are available as carriers for construction of vaccination nanoparticles containing mRNA or recombinant protein antigens. New materials also facilitate a development and use of new technologies for vaccine production (e.g. printing of antigen onto nanofibre-based mucoadhesive film).
- Current and Novel Approach for Influenza vaccines | Influenza Vaccines and Vaccination | Pathogenesis of Influenza Virus | Influenza Viruses -Advance in Detection & Differentiation
Location: Fleming 9
Joseph C. Glorioso
University of Pittsburgh, USA
Institute of Parasitology and Biomedicine, Spain
Czech Academy of Sciences, Czech Republic
Petr Maly is head of Laboratory of Ligand Engineering at the Institute of Biotechnology, Czech Academy of Sciences in Vestec, Czech Republic. He studied at Department of Biochemistry, Faculty of Science, Charles University in Prague, Czech Republic(1980-1985) and completed doctorate at the Institute of Molecular Genetics ASCR (IMG) in Prague. He spent postdoctoral fellowship (1992-1995) at Department of Pathology and Howard Hughes Medical Institute, The University of Michigan Medical School, Ann Arbor, USA, in the laboratory of Prof. John B. Lowe where he published several substantial papers related to in vivorole of mammalian glycosyltransferases. Since 1998 to 2005 he was a research group leader at the IMG in Prague. As a visiting scientist he also worked at Department of Biochemistry and Molecular Biology, College of Medicine, University of Oklahoma, USA. He also was participating investigator of Consortium for Functional Glycomics (USA, 2001–2008) and Member of Editorial Board (2001-2005) and Editor (since 2003) of the Czech journal “Biologicke listy” (Biological Letters). Since 2008, he has been working on the development of combinatorial protein libraries derived from small protein scaffolds andconstruction of novel high-affinity protein binders with therapeutic and diagnostic potential.
Carbohydrates-based immunogens are generally less effective in generation of long-lasting antibody responses and neutralizing epitopes of surface glycoproteins are poorly immunogenic. Therefore, proteins mimicking glycan epitopes represent a promising alternative for development of more protective vaccines. Highly complex combinatorial libraries derived from scaffolds of small and robust protein domains represent an excellent tool for the identification of protein binders mimicking surface glycopeptide epitopes of viruses or bacteria that are recognized by broadly neutralizing antibodies. We use our established concept of a highly complex combinatorial library derived from scaffold of 46 amino acid albumin-binding domain (ABD) and, in combination with ribosome display, we target broadly neutralizing(bn) IgG to identify unique binding candidates recognizing antigen-binding-domain of the tested bn-IgG. In our proof-of-concept study we target glycan epitopes carried by gp120/gp41 protein complex of the HIV-1 Env.ABD variants as potential (glyco)peptide mimetics are currently being characterized for the stimulation of HIV-1 gp120-specific neutralizing antibody response. Thus, ABD-derived recombinant mimotopes could serve as a useful molecular clue for generation of more efficient HIV-1 vaccine and provide a platform for development of other viral or bacterial disease-preventing vaccines.
The project was supported by Czech Ministry of Health grant AZV MZ 15-32198A and Czech Ministry of Education, Youth, and Sport grant CEREBIT CZ.02.1.01/0.0/0.0/16_025/0007397.
Biopharmaceutical New Technologies Corporation, Mainz, Germany
Dr Annete Vogel is presently the Head of Infectious Disease Vaccines, BioNTech AG, Germany. She has been associated with Friedrich-Loeffler-Institut, Federal Reserch Institute for Animal Health as scientist. She has done her education from Georg-August-University Goettingen and completed her DSc from Eberhard-Karls-University Tubingen. She has many publications to her name like “PAR1 contributes to influenza A virus pathogenicity in mice”, “Influenza virus infection aggravates stroke outcome” to name a few. Her Research interest spans around cell culture, animal models, infectious disease, flow cytometry, virology, vaccination, serology and genomics.
Vaccines are the most effective method of controlling infectious diseases. However, todays vaccine production is facing difficult challenges, as the concepts are often not fast and flexible enough to allow quick responses for the efficient control and prevention of global outbreaks of newly emerging and re-emerging viruses and the adaption to new antigenic drifts.
To address this challenge, BioNTech RNA Pharmaceuticals GmbH is developing an innovative synthetic Amplified Immune Response (A.I.R) vaccine platform against Infectious Diseases that is characterized by short manufacturing times, high flexibility and the feasible production of at least 100,000 RNA-based human vaccination doses per week based on the low concentration needed. The administration of in vitro transcribed self-amplifying RNA (saRNA) results in higher antigen expression than delivery of comparable amounts of mRNA, correlating rather to the final subgenomic transcript copy number than to initially transferred RNA amounts. However, antigen expression is still transient as innate immunity effectively prevents persistent replication. Against influenza virus, both B and T cell-responses are induced. We were able to show high antibody titres in mice for over a one year period and protection against live viral challenge after prime-boost as well as after single vaccination by using submicrogram quantities of saRNA. In summary, A.I.R vaccines give equivalent protection against Influenza to mRNA vaccines but at much lower doses.
Novosibirsk State University, Russia
Prokopyeva Elena has completed her PhD at the age of 29 years from FBRI State Research Center of Virology and Biotechnology ‘Vector’(Koltsovo, Russia). Dissertation title: «Phenotypic and genotypic properties of pandemic influenza A(H1N1)pdm09 virus during adaptation to mice of different genotypes». She conduct postdoctoral studies from Novosibirsk State University and Research Center of Experimental and Clinical Medicine (Novosibirsk, Russia). She is the researcher of the Laboratory experimental simulation and pathogenesis of infectious diseases, and the teacher of the course “Embryology” and “ General Histology” at Novosibirsk State University (NSU). She has published more than 25 papers in reputed journals. She has been conducting supervision of undergraduates at NSU since 2016. Also Prokopyeva Elena has obtained different honors and awards in the field of Virology, Pathology and Medical Microbiology.
Pandemic A (H1N1) pdm09 virus has caused substantial morbidity and mortality globally and continues to circulate, which may lead to an increase the pathogenic features of viruses by adaptation to the human. To address this problem, we studied changes of biological properties of the pandemic viruses during adaptation to experimental mammals and analyzed cellular localization of positive-stranded A(H1N1)pdm09 RNA in the inner organs of infected mice. To increase the virulence of pandemic H1N1 isolates in mice, we produced mouse-adapted variants of A(H1N1)pdm09 strain by serial lung-to- lung passages in BALB/c mice. After total of 7 passages we got the lethal strains to BALB/c mice (BALB/c-MA) with meaning of 50% lethal dose 1,2 lgTCID50/ml. Hematoxylin-eosin staining, immunohistochemistry for type A influenza nucleoprotein antigen, and real-time reverse transcription-PCR assay for viral RNA were performed. Complete genome sequences of the wild-type and mouse-adapted A (H1N1)pdm09 influenza viruses revealed 19 amino acid substitutions in different viral proteins (HA, NA, NS2, NS1, PB2, PB1, NP). In lung tissue under the influence of not adapted and mouse-adapted variants of A(H1N1)pdm09 influenza viruses developed interstitial pneumonitis, but it is noted the greatest degree of inflammation in case of infection of the strain BALB/c-MA. Comparative analysis revealed accumulation of viral titers in the brain (3,75±0,22 lgTCID50/ml), liver (2,5±0,5 lgTCID50/ml) and the kidney (0,74±0,48 lgTCID50/ml) in case of infection only of the strain BALB/c-MA. Immunohistochemistry staining of viral antigens was demonstrated in the lung pneumocytes and mucous glands under influence of both wild-type and mouse-adapted viruses. But only in case of infection with BALB/c-MA immunostaining was detected also in the brain, liver, kidney and in the intestine. This study demonstrates cellular localization of positive-stranded A(H1N1)pdm09 RNA in the lungs, brain, liver, kidney and in the intestine that suggests viral replication of the mouse adapted variants of A(H1N1)pdm09 influenza virus in these tissues. The study was supported by a grant from the Russian Scientific Foundation (project No.17-44-07001).