PubMed · 9727479
The source did not provide an abstract. Follow the original record for more information.
Explore connections, maps & timelines
M Cramer. 1998. Experiments using animals.. https://doi.org/10.1097/00006534-199809030-00069
Cite the original work for its findings. Save a collection to share your selection of sources.
KEEP EXPLORING
According to researchers who gathered here at a high-powered summit this month, toxicology may be on the verge of changing the way it collects raw data--adopting a process that could reduce animal use and improve test results. The new approach, called "toxicogenomics," uses DNA arrays to profile gene expression in cells exposed to test compounds. But some leaders in the field warn against rushing too quickly to embrace DNA tests, which are still difficult to interpret.
The European Centre for the Validation of Alternative Methods (ECVAM) facilitates, co-ordinates and participates in validation activities at the European Union level. Various experimental studies, e.g. in the areas of haematotoxicity, reproductive toxicity, nephrotoxicity and epithelial barrier function, and metabolism-mediated toxicity, are underway in ECVAM's laboratories. ECVAM itself is currently involved in the prevalidation/validation of two assays, the colony-forming unit granulocyte/macrophage (CFU-GM) assays for predicting acute neutropenia and the embryonic stem cell test for predicting embryotoxicity. In the areas of metabolism-mediated toxicity and nephrotoxicity and epithelial barrier function, several assays are in the course of development. In many cases, the recommendations of various ECVAM workshops are being followed.
In order to reduce animal testing for quality control of pharmaceutical agents intended for parenteral use, the Limulus amebocyte lysate (LAL) assay is now being accepted in many cases as an alternative to measuring pyrogenic activity of samples in rabbits. However, since the LAL test is specific for cell wall components from Gram-negative bacteria and is sometimes difficult to perform in samples containing large amounts of protein, this alternative still leaves a considerable diagnostic gap. Here, we have optimized a previously established test based on assessing the formation of neopterin or nitrite in interferon-gamma-treated human (THP-1) or murine (J774A.1, RAW264.7) monocytoid cell lines, respectively, in response to bacterial pyrogens. Optimal results were obtained either with THP-1 cells in serum-containing media and using a high concentration of interferon-gamma (IFN-gamma) or with RAW264.7 cells in serum-free media and independent of the IFN-gamma dose. Results were significantly correlated with those obtained by another cell-culture-based assay in which formation of tumor necrosis factor-alpha by THP-1 1G3 cells was assessed. Also in RAW264.7 murine monocytoid cells, formation of nitrite and of tumor necrosis factor-alpha in response to a variety of samples was correlated. Samples shown to be pyrogenic in rabbits in a previous study were unambiguously detected with the test presented here. As expected, the LAL test was negative with cell-free supernatants from Staphylococcus aureus66 kDa). Taken together, these results indicate that the use of monocytoid cell lines and the detection of metabolites which are triggered in the course of immunostimulation could fill the gap left by the LAL test and help to further reduce animal testing for pyrogens.