ࡱ> #`JbjbjmmNN 0008h, 44444      $ h"-Q@-44S~QQQ44QQQ4( `~0S0,###8Q--X   $ 0   0      Poster Abstract Submission Form Alternatives to Animal Testing: New approaches in the development and control of biologicals Deadline: 14th March 2008 APPLICANT OR PARTICIPANT DETAILS Title (Dr., Mr, Mrs, Ms, )DrFirst NameKatjaFamily NameGotovacCompany/InstitutionInstitute of Immunology IncAddress for Correspondence R&D, Rockfellerova 10Town / CityZagrebPostcode / County10 000CountryCroatiaTelephone+385 1 4684500Fax+385 1 4684309E-mailkgotovac@imz.hrPOSTER DETAILS Poster Title: Toward Human Pyrogen Test - possibilities, recommendations and obstaclesFirst Author's Full Name: Tick if the same as above (Dr Katja GotovacFirst Author's Institution, Address, and E-mail (separate using commas) Tick if the same as above (Institute of Immunology Inc, Rockefelerova 10, 10 000 Zagreb, Croatia, kgotovac@imz.hrInitials, last name and institution of all other authors: K Bendelja, V Vdovi, L Lupret Golea, A Vidovi, I Fabijani, A Cepika, I Svoboda Beusan,  Ljevakovi, V `porec, A Gagro, T Sindik Miloaevi All from Institute of Immunology Inc., Zagreb, Croatia ABSTRACT (Please limit to one A4 page, approx. 500 words) The crucial step in the quality control insurance of different pharmaceutical products requires absence of fever-inducing contaminants (pyrogens), especially in drugs intended for the parenteral application. Importance of pyrogen testing is required by EU Pharmacopoeia (Ph.Eur. 2005:2.6.8.). Since 1942, the rabbit pyrogen test (RPT) has been established as a golden standard for testing various pyrogens (Gram-negative endotoxins, non-endotoxin pyrogens, fungal pyrogens). Optionally, the Limulus-Amebocyte-Lysate test (LAL) is used as highly specific test for detection of Gram-negative bacteria endotoxin, but it fails to detect non-endotoxin pyrogens and therefore can not be used solely, instead of RPT. According to recommendations for the development and promotion of tests that will reduce, refine and replace use of laboratory animals, five alternative methods to RPT have been proposed by the European Centre for the Validation of Alternative Methods (ECVAM) recently. These novel in vitro pyrogen tests are based on mechanisms of fiver reaction in humans. The principle of proposed tests is measurement of proinflammatory cytokines (IL-1 and/or IL-6) released from either fresh or cryopreserved human blood cells or a human monocytoid cell line (MM6) in response to the presence of exogenous pyrogens. In 2006 ECVAM submitted results from the study to the Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM) for further evaluation. The ECVAM Scientific Advisory Committee and independent Peer Review Panel, gather in the February 2007 and provided comments, that are in line with ICCVAM draft test method recommendations; (1) proposed tests could serve as a substitute for RPT only for detection of Gram-negative endotoxin on a case-by-case basis, subjected to product specific validation procedure, (2) in vitro pyrogen tests should also include various biological products, and (3) non-endotoxin pyrogens in a future studies. Institute of Immunology Inc., Zagreb is the major regional producers of plasma products, antiserum and viral and bacterial vaccines. Since plasma products are group of biological substances that interfere with the LAL test, they are tested for pyrogens using RPT. Disadvantages of RPT in this case are numerous because most of these products are immunogenic and require a high consumption of animals. Following above stated recommendations, our intention is to introduce and to validate in vitro pyrogen tests for human albumin and intravenous immunoglobulin, using human blood cells and detecting proinflammatory IL-1( productions upon pyrogen (Gram-negative and Gram-positive) stimulation. Results will be compared to standard RPT results and main obstacles from our experience will be discussed. 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