BioVacSafe – Newsletter  July 2013Issue number 2  Editorial: Aldo Tagliabue, EFPIA CoordinatorAt the beginning of April 2009, the news that an influenza strain A/H1N1 coming from pigs was capable of man-to-man transmission came totally unexpected and raised the fear that a new pandemic was starting. In fact, work was still in progress to increase preparedness and control the spread of the avian strain A/H5N1 (endemic in poultry in southeast Asia since 2003). The H5N1 virus could be transmitted by birds to man with low infectivity and high mortality (over 50%), while man-to-man transmission was never proved. The slow diffusion of the avian virus has allowed enough time to create a new generation of vaccines, effective against different influenza strains, with the inclusion of potent adjuvants. These were tested in humans and received the approval from regulatory authorities for use in case of future pandemics. When the swine flu appeared and the risk of pandemics became obvious, it was expected that billions of people would have received in the following months the new adjuvanted vaccines.In those early days of the summer 2009, IMI was beginning the first projects aimed to establish a new way to do research through collaboration between academic institutions and industries. The original IMI strategic agenda did not include projects on vaccines, but the importance of the incoming pandemic flu was the catalytic element that induced to launch a first project on vaccines. Thus the IMI initiative appeared to come at the right moment.  In fact in the last 30 years important progress towards the development of novel vaccines and of novel adjuvants has been observed. The new vaccines against the papilloma virus and against various Neisseria meningitides groups are just two recent examples among several that have been obtained thanks to recombinant DNA and conjugation technologies. Furthermore the first novel adjuvants  have been recently approved after more than 70 years since alum (aluminium potassium solphate) started to be the added component in vaccines capable  of potentiating the immune responsiveness. But novel vaccines always face new challenges in terms of acceptability and safety,  appropriateness to vaccinate at different ages or effectiveness in different geographical areas.  The contest of IMI  public-private partnership appeared as the best approach to  provide a new environment to solve in a transparent and efficient fashion  the complexity represented by the new generation vaccines. Indeed  this challenge was promptly accepted by the 3 major European vaccine companies, and a call for projects on vaccines safety was prepared and released in 2010 by IMI.  Taken advantage of the modern – omics technologies a consortium among excellent academic and industrial partners was created in order to identify  biomarkers that could be indicative of vaccine efficacy and, more important, safety. Still the eternal question regarding the vaccines induced immunity against pathogens remains to be answered. In fact, if the natural disease induces protection against re-infection by causing an inflammatory reaction that brings rubor, tumor, calor, dolor et functio laesa, what is the level of mild inflammation that a vaccine should induce to obtain protection without causing excessive side effects? The project Biomarkers For Enhanced Vaccine Safety ( BioVacSafe) was designed  for addressing this key issue. Aldo Tagliabue EFPIA CoordinatorBioVacSafe Partners ProfileIn every biannual newsletter two of BioVacSafe’s research partners will be highlighted. In this edition you will meet both the University of Surrey in Guilford, UK and Charité – Universitätsmedizin Berlin in Berlin, Germany.   University of SurreyDuring the first 18 months of the BioVacSafe project, partner ‘Surrey’ has made significant progress with its activities in Work Package 1, Task 1.  The University of Surrey’s Clinical Research Centre (CRC) has started its enrolment of healthy volunteers (target n = 120) into three clinical studies that will apply and develop technologies to generate clinical data on inflammation with licensed vaccines as benchmarks, and identify biomarkers to predict acceptable reactogenicity, for correlation with standardised clinical readouts and inflammatory markers assessed in natural infections. Surrey’s Clinical Research Centre (CRC) Team The three clinical studies, employing five different vaccines plus placebo groups, will generate datasets on clinical, metabolic, immunological and gene expression responses as well as a biobank of samples.  Apart from using different vaccines, each study follows the same protocol that will allow data to be combined into one dataset and biobank.  Integrated systems biology analysis of the data and iterative access to the biobank will identify ‘putative biomarkers’ of inflammation, which will be confirmed or refuted in later, larger clinical trials in Gent. The different licensed vaccines are prototypical representatives of a class of vaccine used in a particular target population, and include: Varilrix, Stamaril, Engerix B, Fluad, and Agrippal.  Stamaril, for example, represents a live vaccine given to a population of healthy adults that have no immunity to that disease (Yellow Fever), in the context of a live vaccine.  Varilrix represents a live vaccine given to a population of healthy adults that are already immune to that disease (Chicken pox), in the context of a live booster vaccine.  In each study, regular assessments are performed before and after vaccination, to record clinical events (recorded adverse events), physiological responses (heart rate, blood pressure), metabolic responses (metabolic gene expression), innate immune responses (e.g. cytokine levels), and adaptive immune responses (serum antibody and antigen-specific cellular responses).  Surrey CRC’s extensive experience in running chronobiology studies, with intensive sampling across the 24 hour day under highly controlled conditions, affords a unique opportunity to study inflammatory responses to vaccines by collecting frequent blood samples for multiple analyses.  Whereas previous studies of physiological responses to vaccines have generally obtained single blood samples on Days 0/1/3/7, our studies incorporate a much greater sampling frequency.  During a 7-day residential in-patient stay, incorporating immunisation on the second day, there is extensive sampling both pre- and post-immunisation at 4-12 hour intervals, depending on the study day.  In addition, single blood samples are obtained on days 7, 14, 21 and 28 during outpatient visits.  The selection of the time and frequency of the different sample types, including serum, whole blood, plasma, RNA, PBMCs, CyTOF, etc, has required careful planning to ensure the optimal capture of the timecourse of the response being measured.  In addition, to minimise events caused by factors other than the vaccine, procedures have been developed relating to catheter insertion, removal and monitoring.  The CRC305C study utilises the flu vaccines Fluad and Agrippal.  To eliminate flu infection as a possible confounding variable, Surrey CRC waited until the flu season finished in March/April 2013 before starting the study.  With an expiry date of June 2013 for the Fluad vaccine, this meant there was just 2 months to enrol 48 subjects into the study.  Because of the frequent blood sampling and the labour-intensive sample processing procedures involved, each group had a maximum of 8 subjects.  Six groups were scheduled, plus one ‘straggler’ group in case it was not possible fill all 8 spaces in each group.  An equal number of male and female subjects had to be enrolled.  After the study started, with the clinical, medical, and recruitment teams working virtually non-stop, it was a nail-biting two months before we would find out if we could enrol all 48 subjects before the vaccine expired.  On Friday 31st May, the last three subjects were dosed in the straggler group, just one day before the Fluad expiry date.  With ‘last subject last visit’ for the Fluad and Agrippal study due at the end of June, the medical, clinical and recruitment teams have started to focus on recruitment for the Varilrix and Stamaril study (CRC305A), and to complete the Engerix B study (CRC305B), which started in February 2012. Sample shipment to the different laboratories has begun.  The BioVacSafe partner ‘Max Planck Institute for Infection Biology’ will start to analyse the CRC305C whole blood samples for transcriptomics after the end of June, starting initially with 11 of the 30 sample timepoints per subject.  It will take some time before the results of the sample analyses become available and we are able to finalise the protocols for the larger trials in Gent later in the BioVacSafe project.Blood Sampling Timepoints During The In-Patient Stay And The Outpatient Visits  Charité – Universitätsmedizin BerlinThe Charité is one of the most long-standing hospitals in Germany as well as one of the largest university medical centres in Europe. It enjoys a global reputation as an outstanding training centre. The Charité campus is distributed in four locations across the city of Berlin, comprising over 100 clinics and institutes. In the course of an over three-hundred-year history and its eight Nobel Laureates, the Charité has established a unique reputation. Everyday, a staff of over 13,000 individuals thrives to work according to the 4 key principles: “Research, teaching, healing, helping”. The Charité has achieved outstanding results not only in preclinical research but also in the clinical development of new medicines, medicinal products and other innovations in healthcare. It combines tactful care, scientific acumen, responsibility, respect, and entrepreneurship, all in the context of university medicine and in the interest of a self-determined life.  Foto | Wiebke Peitz | CharitéThe Charité Paediatric Infectious Diseases & Vaccines Team The Charité Paediatric Infectious Diseases & Vaccines Team  (PI: Barbara Rath, MD PhD, contact: kinderinfekte@charite.de) was initiated in 2009.  The multidisciplinary team aims to develop an innovative, comprehensive and personalized approach to the prevention and management of acute infectious diseases in infants and children. The communication with concerned parents and families plays a major role in the daily work of the Paediatric Infectious Diseases & Vaccines Team. Commonly, parents taking their children to an academic medical centre, seek not only medical care, but also evidence-based and detailed information about the safety of anti-infective medications and vaccines. Charité Paediatric Infectious Diseases & Vaccines Team        Foto | Wiebke Peitz | Charité Charité CohortsWith a special focus on vaccine preventable diseases, two quality management programmes have been established in close collaboration with the National Reference Centres for Influenza and Poliomyelitis & Enteroviruses at the Robert Koch Institute, the Department of Microbiology at Charité/ Labor Berlin, the Institute of Mathematics and Computer Science at Freie Universitaet Berlin, Germany, as well as several biotech companies, non governmental organizations and small and medium-sized enterprises in the US, Asia and Europe.The Charité Influenza-Like Disease (= ChILD) Cohort was started in 2009 and encompasses nearly 4000 disease episodes to date. The quality management programme is based in one of the busiest paediatric emergency rooms in Europe. The goal of the programme is the comprehensive disease management and real-time surveillance of influenza and acute respiratory infections in infants and children.The Meningitis Surveillance at Charité (=MenSCh) Cohort, started in 2010 with currently 500 disease episodes, aims to generate a better understanding of the different causes of infections of the central nervous system (CNS) in childhood. This includes the challenging task of detecting occasional, rare autoimmune neurologic disease outcomes following infection versus immunisation.In addition to routine medical care, innovative rapid diagnostic tests are performed at the bedside, in addition to a detailed virologic work-up at the Robert Koch Institute.  Immunisation histories are verified, if available. The clinical presentations are studied in detail, and disease severity is measured in a systematic manner, allowing the identification of disease patterns and progression over time, in collaboration with mathematicians and bioinformaticians at Freie Universitaet Berlin. Foto | Wiebke Peitz | CharitéIn the context of the BioVacSafe Work Package 5, co-lead by Charité as the academic partner, the level of data processing and standardisation is taken to the next level. The common goal of BioVacSafe Work Packages 5 and 6 is to achieve full compliance with data standards developed by BioVacSafe partner CDISC (www.cdisc.org), to achieve a maximum level of data interoperability and meta-analysability across different clinical sites in the Consortium.  To this end, disease area standards and clinical scores will be developed to define the clinical background information required for the selection and classification of specific and unusual disease presentations for the targeted analysis of biomarkers following “natural” infectious disease. In BioVacSafe Work Packages 1, 3, and 4, the Charité cohorts serve as an unbiased, representative tertiary care system to determine the full spectrum of acute infectious disease presentations. This includes influenza and meningitis patients with severe underlying conditions as well as previously healthy subjects, ranging from 0-18 years of age and from outpatients in the ER to inpatients to those requiring intensive care.  The Paediatric and Neonatal Units at Charité are situated in three different Charité campuses in Berlin: an upper-middle class suburb in the south, in the city center, as well as in an ethnically diverse working class neighbourhood in the north. The high level of data standardisation and consistency in the conduct of the Charité cohorts allow for the identification of extreme disease presentations on the background of a perennial hospital surveillance system as well as national sentinel surveillance at the Robert Koch Institute.Targeted biomarker assessments in carefully selected cases will help to set the “upper limits” of biomarker responses in the context of maximal immune system activation during natural infection. This will enable us to distinguish “show-stoppers” from trivial reactions, and contextualise profiles observed in BioVacSafe clinical vaccine trials in the context of natural disease. Foto | Wiebke Peitz | Charité    Project News  BioVacSafe annual meetingThe first annual meeting of BioVacSafe took place in Berlin on the 21st  and 22nd  of March 2013. More than 70 participants from 18 institutions came together both to share and communicate research progresses and to plan the efforts for the next year.Here you can find the Proceedings of the annual meetingUpcoming events:BioVacSafe partners are involved in several meetings, symposia and courses. Interesting to mention are: 3rd International Conference on Vaccine and Vaccination, Embassy Suites Las Vegas, USA July 29-31, 2013. More information can be found here click here to find the program   15th International Congress of Immunology, Milano, Italy 22- 27 August 2013. More information can be found here click here to find the program   7th Vaccine & ISV Congress, Sitges, Barcelona, Spain 27-29 October 2013. More information can be found here   click here to find the program   click here for registration   Advancing Vaccines in the Genomics Era, Rio de Janeiro, Brazil, October 31-Nov 4, 2013. More information can be found here click here to find the program   Recent Pubblications:A list of recent publications related to BioVacSafe project:Maertzdorf J, Weiner J 3rd, Kaufmann SH. Enabling biomarkers for tuberculosis control.  Int J Tuberc Lung Dis. 2012;16(9):1140-8. ReviewWeiner J 3rd, Maertzdorf J, Kaufmann SH. The dual role of biomarkers for understanding basic principles and devising novel intervention strategies in tuberculosis. Ann N Y Acad Sci. 2012 Nov 26. ReviewKaufmann, S. Tuberculosis vaccine development: strength lies in tenacity.  Trends in Immunol. 2012; 33 (7):373-379.Guenounou S, Bosquet N, Dembek CJ, Le Grand R, Cosma A. OMIP-016: Characterization of Antigen-Responsive Macaque and Human T –cells. Cytometry PART A 2013;  83A: 182-184.Kaufmann, S. Tuberculosis vaccines: Time to think about the next generation. Seminars in Immunology, 2013. In press. BioVacSafe contacts:info@biovacsafe.eu – http://www.biovacsafe.eu – Facebook – Twitter – Linkedin This project is supported by the grant n° 115308 from the Innovative Medicines Initiative JU, a joint undertaking between the European Union and the EFPIA companies’ in kind contribution.