Participant Leader: E.L. Roggen

NZ is the world-leading producer of industrial proteins, especially enzymes, with improved performance. Since 1995, NZ has been focussing also on the development of an Immuno-Technology Platform for fast assessment of the immunogenicity of proteins, including therapeutic proteins, by means of cell-based in vitro assays and a proprietary computer-based technique for identification of epitopes in a structural context. NZ is very active at national and international level in the field of Animal Alternatives, and is very strongly in favour of open IPR-free collaborations aiming at validated generally accepted in vitro assays.

Participant Leader: H.-J. Thierse

The department of molecular immunology is an educational and research institute of the university co-associated with the Max-Planck Institute for Immunobiology (MPI) of Freiburg. The research group of Dr. Thierse has a long experience in analysing allergic diseases in human in vitro models as well as in animal models. The research activities focus on early, initial molecular and cellular sensitizing steps in human contact allergy, with primary allergen-interacting proteins from human antigen presenting cells (APC), keratinocytes and allergen-specific T cells. The group contributed significantly to the characterization of allergen-specific human T cell clones and the function of nickel-protein complexes in human nickel allergy, which represents the most common form of human contact allergy in westernized countries. The group also has an established expertise in immunological, proteomic and biochemical techniques, including DIGE (2-dimensional differential gel electrophoresis), multicoloured flow cytometry and confocal microscopy analysis. The laboratory is equipped with up-to-date cell culture facilities, analytical and various in vitro assays. Past and present financial resources include grants from the German Federal Ministry of Education and Research (BMBF), the industry and the local government of Baden-Wuerttemberg.

Including:
a) European Centre for the Validation of Alternative Methods (ECVAM) and
b) European Chemicals Bureau (ECB)

Participant Leader: S. Casati

As general contribution ECVAM will ensure that test methods are developed according to ECVAM's criteria for validation. Specifically, ECVAM will provide the coordination of WP0 and will ensure through ECB, appropriate liaison and involvement of the Competent Authorities experts of the Member States in a) prioritisation of chemicals to be used as training set and b) for the development of a database of fully characterised reference compounds.

Participant Leader: S. Teissier

L'Oréal, the world leader cosmetic company, devotes about 3% of its turnover to research. Since 1984, L'Oréal is actively engaged in the field of Alternatives to Animal testing and participates to validations of in vitro methods. L'Oréal has been involved in previous in vitro European Research Projects based on reconstructed skin models, including or not immunocompetent cells. The Immunotoxicology Group is devoted to the establishment of in vitro methods, predictive of the sensitising potential of raw material. These last years, research has been focused on early, initial molecular and cellular events in human allergy.

Participant Leader: G. Maxwell

The Safety and Environmental Assurance Centre (SEAC), Unilever is the main central resource for the safety evaluation of all ingredients and formulations that Unilever wishes to market anywhere in the world. As such SEAC has a longstanding experience and interest in the field of investigative contact allergy and including the chemistry of skin sensitisation.
Unilever published the Relative Alkylation Index, quantifying the relative extent of a sensitizer binding to skin protein, in 1982 as a function of the dose, the chemical reactivity and hydrophobicity was first by. Since then SEAC has continued to publish on average 1-2 papers a year in this territory. SEAC contributed significantly to the development of the first sensitisation rulebase within the expert prediction system DEREK for Windows (Deductive Estimation of Risk from Existing Knowledge). This is a prediction system containing a large rule base of structural alerts covering many different endpoints but including a substantial number of alerts for skin sensitisation. SEAC has collaborated with the developers of this system in order to enhance it and provide new insights for the development of new or modified rules for skin sensitisation. These new insights have been as a result of PhD studentships funded by SEAC or by significant involvement in the last European Framework V project which focused on fragrance allergy.

Participant Leader: C. Borrebaeck

Department of Immunotechnology has a long-term experience as contractor (BIO2-CT92-0269, BIO4-CT95-0252, BIO4-CT96-0246, PL960389 & BMH4-97-2131) as well as coordinator (QLK3-CT-2000-00270 & BIO4-CT98-0407) of EU projects, in the biotechnological and biomedical area. Furthermore, the department has a long tradition in research based on in vitro cell cultures, molecular biology, protein engineering, genomics and proteomics, the latter two based on microarray platforms. The Swegene Microarray Resource Centre (an academic non-profit service centre) is also located at the department.

Participant Leader: F. Sallusto

The Institute for Research in Biomedicine (IRB) is a private non-profit research institute that conducts basic research in the field of immunology. The laboratory of Dr. Sallusto has been working in the field of human dendritic cell and T lymphocyte biology. In recent years the laboratory has focused on the relationship between migratory capacity and effector function of memory T lymphocytes.

Participant Leader: S. Gibbs

The Department of Dermatology directs its research on the different aspects of immunology and inflammatory diseases. The close link with the clinic ensures the outstanding clinical relevance to patient care. Current research focuses on i) the clinical and pathological aspects of allergic and irritant contact dermatitis and ii) the development and application of tissue engineered skin substitutes for healing chronic wounds and burn wounds. The lab is equipped with all appropriate culture-, cell and tissue-handling facilities, and has immediate access to cell and tissue banks, molecular and micro-array facilities. Our task in this project will be to participate in: i) the identification of in vivo relevant markers for sensitisation, using in vitro derived dendritic cell lines; ii) development and optimalization of (immuno-competent-) 3-dimensional skin / lung model assays; iii) development of in vitro T cell priming assays.

Participant Leader: B.K. Park

The mission of LPOOL is to study drug and chemical toxicity from molecule to man. One of the main aims of the research group is to investigate of the role of metabolism and protein binding in the development of immune-mediated reactions to drugs and chemicals. Over 300 publications have arisen from the group. In order to fulfil the objectives of sens-it-iv, the group has access to state-of-the-art equipment including QToF mass spectrometry, HPLC facilities, flow cytometry facilities, a radiochemical laboratory, class III infection facilities, several real time PCR machines and a proteomics laboratory.

Participant Leader: J. McLeod

UWE has a vibrant research culture, and is widely acknowledged to be at the forefront of science in areas of 3D cell culture models and has wide experience in participating and co-ordinating European projects In-house facilities include full cell culture facilities, FACS, confocal microscope, SEM, TEM, ESEM, proteomics, transcriptomics, 2-D electrophoresis and differential gel electrophoresis (DIGE, Amersham Bioscience), gene cloning, micro-array facilities and quantitative RT-PCR, and both matrix assisted laser desorption ionisation - time of flight (MALDI-TOF-MS) and electrospray ionisation - liquid chromatography mass spectrometers (ESI-LC-MS).

Participant Leader: A. Braun

The Department of Immunology and Allergology conducts pharmacological efficacy tests for research in allergy, inflammation and infection as well as different immunotoxicity tests. The target organ of most of these studies is the lung.. All tests can be performed according to the quality requirements of GLP. For registration purposes (EMEA, OECD, FDA), both the pharmacological efficacy and the allergenic or toxicological potential of substance candidates are determined. Immunological endpoints can be identified both in in vitro test systems and in animal models. We have access to a broad range of "state of the art" analytical methods including immunological, molecular-biological, histological and lung physiological methods.

Participant Leader: M. Marinovich

The Department of Pharmacological Sciences is part of the Faculty of Pharmacy of the University of Milan, Italy. It has about 140 people dedicated to research and teaching. The laboratory of "Immunotoxicology and Immunopharmacology" is part of the Laboratory of Toxicology, directed by Professor Corrado L. Galli. The overall interest of the Laboratory of "Immunotoxicology and Immunopharmacology" is on:
- Skin immunotoxicity (irritation and allergy),
- immunosenescence,
- Effects of pesticides on immune system of occupational exposed workers

In vitro methods
1. Cytotoxicity: LDH release, MTT reduction, protein synthesis, apoptosis, cell cycle, mitochondrial potential, ROS production, calcium measurement
2. Immunotoxicity: phenotyping, cytokine production, cell proliferation, NK cell activity, oxidative burst, whole blood assay, transcription factors activation, DNA macroarray
3. Cell culturing: peripheral blood leukocytes, primary macrophages (alveolar, splenic and peritoneal), HEL30 and NTCT keratinocyte cell lines, THP-1 cells, fibroblast, reconstituted human epidermis (EpiDerm, EpiSkin)
4. Other relevant matters: our main interest in the project is the development of alternative in vitro testing to discriminate allergy from irritation based on differential gene expression and, the understanding of the role of the new marker(s) in the induction of skin irritation and allergy.

Participant Leader: I. Nelissen

The strategic research within the Environmental Toxicology Unit (TOX) of Vito focuses on the development and application of in vitro technologies that aim to reduce, replace and refine the use of animal testing. Our expertise extends in the domains, genotoxicology, endocrine disruption and haemato-immunotoxicology. A considerable part of our personnel is dedicated to immunotoxicity research and works on the development of an in vitro assay for contact and respiratory sensitisation. Within the Sensi-it-iv project we will contribute to WP2 and WP8 and focus on the CD34-dendritic cell model and on cell lines such as the A549 epithelial cell line, TOX has ample infrastructure for cell culture, protein analysis (FACS, ELISA) and molecular biology (real-time quantitative PCR, microarray analyses).

Participant Leader: M. Assenmacher

For more than ten years the research and development department at Miltenyi has a large programme to develop innovative T cell and dendritic cell (DC) isolation reagents, culture systems and analysis tools. We will contribute with our unique expertise on cell separation and analysis to WP3 (Dendritic cell - T cell interaction). In particular on the characterization of allergen-specific T cells as well as on the ultimate aim, the development of an assay to assess antigenicity. To this end we will contribute to create defined compositions of DCs and T cells, to optimize the culture system (including attempts to enrich for antigen-specific T cells) and to the sensitive detection and characterization antigen-specific T cells.

Participant Leader: A. Cavani

The IDI hospital represents the largest Dermatological hospital in Italy, with more than 200 beds and 500 daily outpatients, and has been qualified as IRCCS (Institute for Biomedical Research and Care) since 1993. The laboratory of Immunology is mainly focused on the identification of pathomechanisms responsible for immuno-mediated skin diseases, allergic contact dermatitis to small chemicals in particular, being connected with a large allergology department, that can provide blood samples from patients affected with hapten-induced diseases.
The Laboratory is fully equipped for immunology, cellular and molecular biology research, including cell culture facilities, light, electron and confocal microscopes, flow cytometers, cell sorter, spectrophotometers, realtime PCR, electrophoresis and blotting apparatus, ELISA and ELISPOT readers.
Major contribution to the project of the Laboratory of Immunology at IDI will be to characterize T cell populations involved in human allergic contact dermatitis to chemicals, to develop DC-T cell and EC-DC-T cell based predictive assays.

Participant Leader: H.U. Weltzien

The MPI IB is a nationally and internationally recognised and leading Institute in the field of basic research in Immunology and developmental Biology. H.U.Weltzien's laboratory has significantly contributed to the understanding of the structural, molecular and cellular basis of antigen recognition by T cells. Internationally particular recognised is their work on T cell interactions with chemical haptens, which forms a basis for modern understanding of the mechanisms underlying chemical-induced allergies. More recent work has concentrated on mechanisms involved in the activation of human T cells from allergic individuals allergic to penicillins or heavy metals.

Participant Leader: E. Maggi

The expertise of the laboratory is mainly devoted to the fields of immune regulation, functional T cell subsets, cytokines and chemokine networks applied to atopy, immunodeficiencies and autoimmunity. The research fields of the group: i)Characterization of allergens and T cell response to them, ii) Regulatory mechanisms of IgE response in humans, iii) Paradigm Th1/Th2, iv) Role of chemokines and chemokine receptors in thymic lymphopoiesis and tumorogenesis, v) Regulatory T cells of the immune response.
The group employs the cellular and molecular biology techniques relevant to the present project such as: Lymphocyte culture and T cell cloning procedures, production of antigen(peptide)-specific T cell lines, development of in vitro models for studying the modulation of Th cell differentiation, ELISA or biological assays for cytokine and chemokine measurement, Western Blot analysis for membrane molecules, quantitative real-time RT-PCR (TaqMan), multiparametric cytofluorescence analysis, cytofluorometric evaluation of intracellular cytokine at single cell level, cytoflurometric cell sorting, DNA sequencing, double immunohistochemical staining of cells, in situ hybridization for cytokines/chemokines or their receptors, transient and permanent transfection of many types of cells, confocal microscopy analysis.

Participant Leader: G. Renner

Colipa will contribute to the Technology Transfer/Dissemination activities of the project through a well-established membership platform of companies and national cosmetic associations in the EU. Its Steering Committee on Alternatives to Animal Testing (SCAAT) coordinates a collaborative research program on skin sensitization. Colipa will contribute to Sens-it-iv's scientific integration with the SCAAT program, in order to accelerate progress, optimise applicability and promote adoption of the final cell-based methods by a large part of the end-users.

Participant Leader: P. Aeby

Cosmital SA is a subsidiary of Wella AG, Darmstadt. One of the main focuses of Cosmital SA is the research and development of in vitro alternatives to animal tests for the evaluation of the toxicological profiles of chemicals.

Participant Leader: Dr. Joan Albert Vericat

IVTIP was established in 1993 to serve as an industrial forum following progress in the development of in vitro testing as applied to regulatory testing and in the compound discovery and development process. Currently it has 31 members; its members are European cosmetic, chemical and pharmaceutical companies, as well as dedicated SMEs operating in technology niches. Of its members, 40% are SMEs.
During the past 11 years, IVTIP's efforts have resulted in

• The building of a large network between industry and academia operating in the field of in vitro testing. Through its meetings and its members, it has succeeded in creating a personalized network involving more than 60 European companies and close to 100 dedicated scientists;
• Successful examples of technology transfer. Through its meetings, most of the EU funded research projects in in vitro testing and the 3Rs have been followed, its coordinators invited to present their results and links established between IVTIP member companies and academics. Furthermore, IVTIP initiated the Technology Transfer Review Project, an EU funded project (QLK3-1999-30005), which served as a template for the construction of this project. The Review Project demonstrated the usefulness of in depth analysis of a project's results;
• Increased use of in vitro testing approaches in European industry, both for regulatory testing as well as for use in proprietary compound selection and development processes.

IVTIP has also advised the European Commission and the European Parliament on industry's needs in the area of in vitro testing research through regular position papers that reflect the consensus opinion of its member companies.

Participant Leader: W.P. Hermans

Scientific Writing & Consultancy is a private partnership, established in 1985 by its current director, Dr. W. Hermans. The partnership is now located in Pedreguer, Spain and registered as a Sociedad Civil. Previously it was located in the Netherlands (1985-1994) and Ireland (1994-2004).
Since its foundation, the partnership is specialized in the writing of original brochures, books and leaflets about science and technology (specialization biotechnology) intended for a well educated but non-specialized public. Since 1985, more than 50 brochures and booklets have been researched, written and published, all based on original sources (scientific articles, interviews with scientists).
The consultancy part consists of the running of the Secretariats of two industrial platforms: ACTIP, the Animal Cell Technology Industrial Platform and IVTIP, the In Vitro Testing Industrial Platform. These are platforms where representatives of European industries meet, and where recommendations for EU research policy are formulated. These recommendations are submitted to the European Commission and the European Parliament. Dr. Hermans is Executive Secretary for these two platforms: since 1990 for ACTIP and since 1993 for IVTIP. She takes care of day to day business, formulates policy, and together with her staff organizes 4 plenary 2-day meetings per year where representatives of member companies discuss business issues and meet researchers to talk about frontline research projects. The activities within these platforms take the form of transfer of technology and use of RTD results.

Participant Leader: H. Merk

The focus of this research group relates to probing the mechanisms of allergic reactions in the skin to small molecular weight compounds capable of causing disorders such as allergic contact dermatitis and drug allergy. The scientific members are multi-disciplinary and include physician-scientists including dermatologists and allergologists as well as basic scientists in molecular biology and immuno-biology. For more than 20 years the participant has been investigating the mechanism of extrahepatic xenobiotic metabolism in the skin and in immunocompetent cells and its role in evoking sensitization against small molecular weight compounds. All of the investigators involved in this project are members of the Arbeitsgemeinschaft Dermatologische Forschung (ADF), Deutsche Dermatologische Gesellschaft (DDG) and the European Society of Dermatology Research (ESDR). The unit has access to state-of-the-art equipment such as microarray workstation (hybridisation, scanner), flow cytometry facilities, several real time PCR machines, sequencers, a radiochemical laboratory and cell culture laboratories (Keratinocytes, lymphocytes, dendritic cells, 3-D skin equivalents).

Participant Leader: J.P. Lepoittevin

The mission of the research group is to study relations between the chemical structure of a molecule and its ability to act as a skin sensitizer. The participant's interest in the field of chemical allergy spans over 20 years. One of the main aims of the research group is to investigate how haptens and pro-haptens react with proteins to form new antigenic structures. These studies are carried out at the molecular level allowing to precisely identify the mechanism by which haptens are reacting with each modified amino acid. Over 100 publications have arisen from the group, the activities of which are available on a web-site (http://www-chimie.u-strasbg.fr/~ldc/). The research group has been involved in several EU projects related to skin allergy and development of alternative methods (BIOT-CT90-0186-C, Biomed II BMH4-CT96-0877, QLK4-1999-01558). In order to fulfil the objectives, the group has access to state-of-the-art equipment including high field NMR spectroscopy, mass spectrometry, HPLC facilities as well as synthesis of labelled molecules and peptide synthesis. For this project, newly refurbished laboratories dedicated to bioorganic chemistry will be used.

Participant Leader: J. Riera

Proteomika works in the field of biomedical research using proteomics tools for target discovery with a strong interest toxicoproteomics The company is a member of the Human Proteome Organisation (HUPO) and is highly experienced in sample preparation technologies. The company uses both 2-dimensional gel electrophoresis and chromatographic techniques for protein and peptide separation and uses MALDI-TOF, TOF/TOF and Q-TOF mass spectrometers for high throughput protein identification and characterisation of post-translational protein modifications.

Participant Leader: C. Helma

The development and application of Data Mining and Machine Learning techniques for toxicologial problems is one of the main research areas of the Machine Learning Lab at the Institute for Computer Science (ALU-ICS). The present research focus is the development of inductive databases (i.e. databases that can be queried for patterns and regularities within the data) for various biological and chemical domains (e.g. DNA and Protein sequences, 2D and 3D molecular structures,...) The lab is widely acknowledged to be one of the driving forces in the development of novel techniques for the prediction of the toxicity of untested compounds (e.g. MolFea and lazar). Staff from the Machine Learning Lab were the main organizers of the Predictive Toxicology Challenge 2000-2001, editors of a special section about Predictive Toxicology for the journal Bioinformatics and editor of a recent textbook about Predictive Toxicology. The Machine Learning Lab has a rich experience in European Projects. The chair (Prof. DeRaedt) coordinated two ESPRIT projects on Inductive Logic Programming (ILP1 and ILP2) and was one of the principal investigators of the ALADIN (ESPRIT-28.623) project. Presently the lab is involved in the CINQ (IST-2000-26469) and APRIL (IST-2001-33035) projects and coordinates the DAISY (HPMT-CT-2001-00251) Program. Apart from that it is involved in co operations with the industry and organized the 12th European Conference on Machine Learning and the 5th European Conference on Principles and Practice of Data Mining and Knowledge Discovery in Databases. The Machine Learning Lab will take care of the Data Management and Bioinformatics part of the project. It will provide a central repository for experimental data from the Sens-it-iv project, a web-interface for the encrypted submission and retrieval of data and possibilities to identify relevant patterns and relationships (e.g. similarities of in-vitro cell lines) within the data.

Participant Leader: S.F. Martin

The Department of Dermatology at the Faculty for Medicine in Freiburg has a great expertise in allergy diagnosis, allergy testing and treatment including hyposensitization therapy. Not only do we perform ambulant therapy, but the associated clinic treats a large number of stationary patients. The research laboratories are specialized in studying the immunologic pathomechanisms of allergic and autoimmune diseases of the skin. The strength of this setting is the in-house collaboration of clinicians and researchers involved in basic science. The specific expertise of Dr. Martin's laboratory is the 15-year experience in the analysis of T cell responses to chemical allergens including recently also plant allergens with a focus on contact hypersensitivities. The laboratory is investigating the role and interplay of effector and regulatory T cells, dendritic cells and the programming of T cells for skin specific migration by dendritic cells and peripheral and lymphatic tissue microenvironments. The laboratory has all relevant facilities to perform tissue culture, functional assay and phenotyping, protein biochemistry and molecular biology as well as radioactive assays. A core facility within the Faculty provides proteomic and genomic platform technology.
Our work benefits from the outstanding infrastructure in Freiburg with a strong interactive community of clinicans and scientists involved in allergy and immunology research. This structure resulted from the establishment of a Clinical Research Group "Pathomechanisms of the Allergic Inflammation" which was funded by the German Ministry for Education and Research (BMBF) from 1.1.1998 until 31.12.2003. The Faculty for Medicine is now funding a core group at the Department of Dermatology and the continuation of a growing research group is actively pursued and supported by the Faculty. Further partners are the Department of Pneumology, the Department of Pediatrics, the Faculty for Biology, the Faculty for Pharmacology and Toxicology and the Max-Planck-Institute for Immunobiology. This extraordinary setup allows us to use the expertise of the other partners in the form of active collaborations.

Participant Leader: Vera Rogiers

ecopa (European Consensus-Platform for Alternatives): This is the only quadripartite not-for-profit organisation which promotes a consensus strategy for 3R-alternative methods in the EU. Innovative is that the 4 parties having a major interest, animal welfare, industry, government and academia, are equally represented and reach consensus. The concept of consensus between the 4 parties concerned has been accepted in various countries as an efficient way to stimulate research into alternatives to animal experiments and enforcing the acceptance of alternatives in experimental practice. ecopa's goal is to respond to the need for the creation of a pan-European platform. Besides of the fact that a link is needed between the different national platforms, consensus discussions with all relevant groups will maximise results and minimise conflicts within the 3 R's strategy. Jointly accepted opinions transformed into a strong plea have substantial impact for progress on issues related to alternative methods.
ecopa has co-initiated and currently supports among others the CONAM project under the EU 6th Framework Program, whose main objectives are to build a solid network on 3R-alternatives, ideally including all European countries and with the aim to deliver critical consensus expert opinions on 3R-issues, to draw attention on new alternatives and technologies, to disseminate this information and to initiate collaboration. An information system on alternative method development has been created for this purpose for the first time in Europe that will, almost in real time, supply the interested organizations, institutions and individuals within EU and non-EU candidate countries with the respective information.
Moreover, ecopa's involvement in the ReProTect and the PREDICTOMICS 6th Framework Program Integrated projects indicates their focus, expertise and commitment in the development and promotion of alternative methods. ecopa's networking offers equilibrated, scientifically-sound and technically-relevant expertise which can additionally benefit the technology transfer and dissemination objectives of the Sens-it-iv project.