BioVacSafe – Newsletter  January 2014 Issue number 3 BioVacSafe’s  second year progress.On March 1st 2012 the IMI-JU funded project – Biomarkers for Enhanced Vaccine Safety (BioVacSafe) – initiated its collaborative work to develop new tools, methods and guidelines for the evaluation of vaccine reactogenicity and enhancing immunosafety of novel vaccines.During the second year of the BioVacSafe project, great progress has been made in the development of  cutting edge tools to speed up and improve the testing and monitoring of vaccine safety. Clinical trials have been successfully started and the analysis of samples to measure inflammatory biomarkers is proceeding with success. Pre-clinical studies are successfully progressing to define immune activation and inflammatory biomarkers in response to vaccination and infection. The main achievements of the project during the second year will be presented and discussed at the BioVacSafe Annual Meeting that will be held in March in Siena and more information about the project progress will be provided in the next newsletter. BioVacSafe Partners ProfileIn every biannual newsletter the profile of some  of BioVacSafe’s research partners is highlighted. In this edition you will meet the CEA -Division of Immuno-Virology in  Fontenay-aux-Roses, France; the HPA-NIBSC in  South Mimms, UK and CDISC in Austin, TX, USA.    Division of ImmunoVirology-CEAThe Division of Immuno-Virology of the Institute of Emerging Diseases and Innovative Therapies (iMETI) develops non-human primate (NHP) models of interest for the national and international community, for the testing of vaccines, microbicides and treatments against several human pathogens. The division has acquired strong expertise and skills in viral pathogenesis, chemo-prophylaxis (microbicides, pre- and post-exposure prophylaxis) and vaccines using NHP models of human infections, particularly through its contribution to the development of NHP model of human HIV infection and AIDS. Activities have been now extended to several important human diseases including tuberculosis and malaria.In 2013, the Division of Immuno-Virology, the Institut Pasteur, the “Institut National de la Santé et de la Recherche Médicale” (INSERM), the “Agence Nationale de Recherche sur le SIDA et les Hépatites Virales” (ANRS), Université Paris Sud 11 (UPS-11) and Bertin Pharma have decided to join forces to create a research center for “Infectious Disease Models and Innovative Therapies” (IDMIT) at the CEA site in Fontenay-aux-Roses.  In this new entity, industrial and academic partners can benefit from a unique infrastructure including state-of-the art and extended NHP facilities and outstanding equipments with highly advanced technologies for monitoring infections, host response, treatments and vaccines efficacy: genomics, flow cytometry, mass-cytometry and in vivo imaging.Cytometry by Time of Flight (CyTOF) or mass cytometry was recently implemented at the FlowCyTech core facility, one of the core facilities that support the Division of Immuno-Virology.In the context of the BioVacSafe project partner CEA, with its FlowCyTech team,  is involved in Work Package2 “Establishment of reliable in vivo animal models and in vitro models predicting early inflammation and autoimmune diseases” and will conduct CyTOF analysis on samples of the Biovacsafe Clinical studies. To this aimCEA established a special experimental procedureto allow collection of blood samples at Surrey’s Clinical Research Center and shipping at the Division of Immuno-Virology for CyTOF analysis. The experimental procedure allows freezing cellular component of the blood with preservation of all its components including the granulocytes that are usually lost during freeze and thaw protocols. Samples, once shipped at the Division of Immuno-Virology will be stored in the biobank for successive CyTOF analysis. This procedure will be used to study innate responses in the “CRC305 Varilix and Stamaril” BioVacSafe pilot human trials. “Crédits photos : P.Stroppa/CEA”Mass cytometry address most of the limitations of the classical fluorescence-based flow cytometry. The key innovations are a different chemical nature of the labels and a non-optical detection system based on mass spectrometry. Antibodies are conjugated to metals, specifically, transitional elements isotopes normally absent in biological systems. These conjugates are used as in other flow-cytometry based protocols to label cells of interest. Cells labelled with metal tagged antibodies are then, vaporized in a spray chamber and delivered as single cells to a plasma chamber where they are atomized and ionized. Each cell originates a cloud of ions that is analyzed by time of flight (TOF) mass spectrometry. Since, the elements conjugated to antibodies are not naturally found in biological samples the signature of each metal clearly represents the labelling of a single cell by a specific antibody. Technically speaking the mass cytometer combines a high temperature inductively coupled plasma (ICP) source with detection by time of flight mass spectrometry (TOF-MS). The main advantage of this innovative technology is the absence of overlap among different detection channels. Indeed, each transitional metal used to label the antibodies, is uniquely detected and identified according to its molecular weight. The system is able to detect metals with a molecular weight comprised between 125 and 215 kilodaltons, and 34 different metals can be detected simultaneously with the actual set of reagents. In the future, it will be possible to analyze simultaneously 100 different metal tags. Importantly, and differently from flow cytometry, the chemical properties of the metal tags are quite similar and, therefore there is not an effect of the tag on the staining results. Furthermore, the use of mass spectrometry as read-out system allows for absolute quantification.In order to study the innate immune response a panel of 31 different monoclonal antibodies has been established. Data will be analyzed with different flow cytometry software specialized for multi parameter analysis. Principal component analysis and cluster analysis will be mainly used to avoid the subjective procedure of supervised gating. All generated results will be made available to the BioVacSafe partners via a common database were CyTOF data will integrate assays performed by other laboratory. We hope the integrated analysis this high multiparameter dataset will shed light on interesting markers of vaccine safety. “Crédits photos : P.Stroppa/CEA”    HPA-NIBSCThe National Institute for Biological Standards and Control (NIBSC) is part of the Medicines and Healthcare Products Regulatory Agency (MHRA).  The MHRA is an executive agency of the Department of Health that is responsible for the regulation of all medicines, medical devices and equipment used in the UK by ensuring they work and are acceptably safe; and the investigation of harmful incidents. The MHRA also looks after blood and blood products, working with UK blood services, healthcare providers, and other relevant organisations to improve blood quality and safety. NIBSC is a global leader in standardisation and control of biological medicines. As the leading WHO International Laboratory for Standards, NIBSC develops and produces over 90% of international standards. In addition, NIBSC is the Official Medicines Control Laboratory (OMCL) for the UK, and is therefore responsible for performing batch release testing of biological medicines including blood products and vaccines for the European market. To support these two functions, considerable research and development (R&D) efforts are being undertaken at NIBSC. We also have an established track record in developing novel quality control assays and managing the processes through to assay qualification and validation within a regulatory environment. Moreover, scientists from NIBSC often lead the establishment of recommendations and guidelines for WHO and those regulatory documents for inclusions in the European Pharmacopoeia, European Medicines Agency and the like.  As part of BioVacSafe Work Package 6 “Guidelines, standards, classifications and harmonization”, the NIBSC is involved in (1) harmonisation and standardisation of preclinical and clinical tests used to evaluate biomarkers for the safety of vaccines identified by the work packages of the BioVacSafe project, including the preparation of standard materials for use in the tests; and (2) qualification of the identified biomarkers and assays used to measure the biomarkers, as well as the preparation and standardization of guidelines that are the outcomes of the qualification process. NIBSC will, in collaboration with BioVacSafe partners, formulate and characterise potential reference materials as well as establish the reference reagents for use in biological assays within the consortium. At the NIBSC, there is decades of skilled expertise in the processing of a wide range of biologicals to exacting specifications (including accurate fill precision/ low coefficient of variation, low residual moisture for lyophilised products). Customised standards, tailored for a particular biological medicine or in vitro diagnostic test can be developed at the NIBSC. The institute’s expertise in formulation helps to ensure batch homogeneity and stability. In addition, standards can be liquid filled or filled and freeze-dried (at very low residual moisture levels) into a range of ampoules, vials and tubes for both non-infectious materials and materials of up to level 3 microbiological containment. Furthermore, extensive analytical services are available to monitor the quality of the final material once filled. The key players from NIBSC within the BioVacSafe consortium are Dr Mei Mei Ho and Dr Omodele (Dele) Ashiru. Dr Mei Mei Ho is the Principal Scientist and Group Leader of the Immunology and Cellular Immunity Group within the Bacteriology Division at NIBSC. The group focuses on R&D, quality control and standardisation of biological medicines used in tuberculosis (TB) and malaria fields. Notably, the group has expertise in the immunology and pathology of pulmonary TB, together with an established murine aerosol challenge model for evaluation of new TB vaccine candidates. The quality control and standardisation activities are accredited to ISO/IEC 17025 (for EU batch release) and 9001 respectively. The group is one of the key partners in TRANSVAC, an EC-funded consortium (2009-2013), working on the harmonisation of immunological assays (including ELISpot, flow cytometry – intracellular cytokine staining and cytokine ELISA assays), and is also responsible for the development and provision of reference standards/ reagents to scientific research communities within EU. Together with the Centre For Aids Reagents (CFAR), Mei’s group is coordinating and leading an EC-funded consortium (2013-2017), EURIPRED, to create partnerships between European scientists and international research teams from disease endemic countries, building an infrastructure for exchange, selection, development, production and validation of reference standards/ reagents, assays and training provision in the field of poverty related diseases R&D.Dele joined the team in October 2013 as a senior scientist; a role funded by the BioVacSafe project. She has a research background in molecular viral immunology, with a particular focus on the innate immune system. She has several years of experience in immunology, virology, molecular biology and biochemistry techniques, as a result of several postdoctoral research posts undertaken at the University of Cambridge, University of Strasbourg and University of Oxford. Since working on the BioVacSafe project, Dele has been liaising with Work Package 2 partners in order to facilitate the development of reference standards and assays in the influenza area.  C-DISCCDISC is a global, open, multidisciplinary, non-profit organization that has established standards to support the acquisition, exchange, submission and archive of clinical research data and metadata. BioVacSafe is the 2nd IMI consortium project in which CDISC (Clinical Data Interchange Standards Consortium) was invited to be a partner organization, which helped provide the impetus for formation of the CDISC Europe Foundation in Brussels. IMI and CDISC signed a memorandum of understanding in 2011. CDISC has worked with global pharmaceutical research stakeholders for many years. BioVacSafe, however, has provided the opportunity for closer collaboration with European vaccine manufacturers, key academic institutions and clinical experts in vaccine-preventable diseases across Europe. Under the BioVacSafe project CDISC will contribute to Work Programme 5 “Biovacsafe Data Management System and AEFI Assessment “.For nearly 15-years, CDISC has developed and published global data standards for us across the clinical research process and data chain from protocol…to data collection…to tabulation and analyses…and ultimately regulatory submission and reporting. As depicted in the diagram, CDISC core standards include:PRM:  Protocol Representation Model for study protocols and designCDASH:  Clinical Data Acquisition Standard for CRF data collectionADaM:  Analysis Data Model for analysis datasets SDTM:  Study Data Tabulation Model for human clinical trial data to regulatorsSEND: Standard for Exchange of Non-Clinical Data for animal toxicity dataBRIDG Model: provides an overarching UML model for all CDISC standards and provides a connection between clinical research and healthcareUnder the BioVacSafe project CDISC CDASH standards for data collection and controlled terminology have been implemented at Charité and the University of Surrey.CDISC has also developed and published standard controlled terminology codelists primarily supporting CDASH, SEND and SDTM. More recently, over the past couple of years CDISC has been developing additional data standards specific to therapeutic areas (TA) such as Alzheimer’s, Asthma, Cardiovascular Disease, Diabetes, Kidney Disease, Parkinson’s and Tuberculosis with many additional TA projects coming online in 2014 and years following, including for vaccine preventable diseases such as influenza. Under WP5 CDISC-compatible CRFs were discussed and generated during the first 6 months of the project to ensure consensus across BioVacSafe clinical trial and cohort sites and to ensure data is collected in a consistent way. Additionally, in year 2 the Charité database and data elements were harmonized and mapped to the CDISC BRIDG Model and conversion / mapping to SDTM ver1.4 and SDTM Implementation Guide ver3.2 is ongoing. This work comprises the initial steps toward development of the full BioVacSafe Data Standards Package. Working with key BioVacSafe stakeholders under WP5, a CDISC Vaccine Data Standard and User/Implementation Guide vaccine will be developed, using real life data at Charité as a starting point. The User/Implementation Guide describes the most common research concepts relevant to vaccine studies and the necessary metadata to represent such data consistent with CDISC standards, providing guidance on implementation of SDTM to represent vaccine data in regulatory submissions. This work will form the basis of a global vaccine standard as well as additional TA standards for vaccine preventable diseases. Once developed the new vaccine data standard will undergo the CDISC global public review process before becoming open and freely available as a tangible outcome of BioVacSafe, accepted and endorsed by regulatory agencies. The data standard will be in compliance with regulatory requirements in the US, Europe and Japan. In June 2013, The EMA released draft policy for Publication and Access to Clinical Trial Data as part of the EMA’s Clinical Trial Transparency Initiative. The draft policy states – “In future, CDISC shall be the required standards, in line with future guidance from the Agency”. EMA plans to release final policy in early 2014. Similar position statements have been released by PMDA (Japan) and US FDA, indicating requirement of CDISC standards in regulatory submissions by 2016 and 2017 respectively.In summary, BioVacSafe data standards implementation and development under WP5 is progressing well and will result in on time delivery of a full data standards package for vaccine information. This will ensure consistency of data across BioVacSafe and facilitate vaccine research projects in the future since the new standards will streamline data flow from collection through analysis and reporting to regulatory agencies.  BioVacSafe 2nd Annual MeetingThe BioVacSafe annual meeting will be held on 20-21 March 2014 in Siena.  All partners of the project are invited for this meeting – that will be attended by around 70 participants – to present and discuss the results of the second year of research (by invitation only). Upcoming events:BioVacSafe partners are involved in several meetings, symposia and courses. Interesting to mention are: Omics Meets Cell Biology: Applications to Human Health and Disease February 18—23, 2014, Sagebrush Inn and Conference Center, Taos, New Mexico, USA.  More information can be found hereNew Horizons for Vaccine Research and Innovation, 12-13 March 2014, Brussels, Belgium.  Biomarker Summit 2014, March 19-21, 2014, San Diego, CA. More information can be found hereBiomarkers & Diagnostics World Congress 2014,Cambridge Healthtech Institute’s Tenth Annual,  April 30-May 2, 2014, Loews Philadelphia Hotel, Philadelphia, PA.  More information can be found here  World Vaccine Congress Asia 2014, 9-12 June 2014, Suntec  Singapore convention & exhibition centre. More information can be found here  8th Vaccine & ISV Congress, 26-28 October 2014 | Philadelphia, PA, USA. More information can be found here click here to find the program  World Vaccine Congress 2014, 22-23 October 2014, Lille grand Palais, Lille, France. More information can be found here  The Modes of Action of Vaccine Adjuvants, October 8—13, 2014 Sheraton Seattle Hotel, Seattle, Washington, USA. More information can be found here 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. http://www.ncbi.nlm.nih.gov/pubmed/22871324Weiner 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. 2013; 1283: 22-29. http://www.ncbi.nlm.nih.gov/pubmed/23181737Kaufmann, S. Tuberculosis vaccine development: strength lies in tenacity.  Trends in Immunol. 2012; 33 (7):373-379. http://www.ncbi.nlm.nih.gov/pubmed/22560865Guenounou 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. http://onlinelibrary.wiley.com/doi/10.1002/cyto.a.22233/abstractKaufmann, S. Tuberculosis vaccines: Time to think about the next generation. Seminars in Immunology, 2013; 25: 172-181. http://www.sciencedirect.com/science/article/pii/S1044532313000225A. R. Everitt, S. Clare, J. U. McDonald,, L.Kane, K.Harcourt, M. Ahras, A. Lall, C.Hale, A. Rodgers, D. B. Young, A. Haque, O. Billker, J. S. Tregoning, G. Dougan, P. Kellam:  Defining the range of pathogens susceptible to ifitm3 restriction using a knockout mouse mode. Plos one 2013; 8: 1- 12. http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0080723Kaufmann, S.H.E., A. Dorhoi: Inflammation in tuberculosis: interactions, imbalances and interventions. (Special issue: Host pathogens). Curr. Opin. Immunol. 2013; 25: 441–449. http://www.sciencedirect.com/science/article/pii/S0952791513000757Kaufmann, S.H.E., J. Weiner 3rd: Recent advances towards tuberculosis control: vaccines and biomarkers. J Intern Med. 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.