title: Evaluation and prevalidation of an immunotoxicity test based on human whole-blood cytokine release.
abstract: Immunotoxicology is a relatively new field in toxicology, and is one of emerging importance, because immunotoxicity appears to contribute to the development of cancer, autoimmune disorders, allergies and other diseases. At present, there is a lack of human cell-based immunotoxicity assays for predicting the toxicity of xenobiotics toward the immune system in a simple, fast, economical and reliable way. Existing immunotoxicity tests are mainly performed in animals, although species differences favour human-based testing. Whole-blood cytokine release models have attracted increasing interest, and are broadly used for pharmacological in vitro and ex vivo studies, as well as for pyrogenicity testing. We have adapted those methods for immunotoxicity testing, to permit the potency testing of immunostimulants and immunosuppressants. Following stimulation with a lipopolysaccharide or staphylococcal enterotoxin B, monocytes and lymphocytes release interleukin-1beta and interleukin-4, respectively. Thirty-one pharmaceutical compounds, with known effects on the immune system, were used to optimise and standardise the method, by analysing their effects on cytokine release. The in vitro results were expressed as IC50 values for immunosuppression, and SC(4) (fourfold increase) values for immunostimulation, and compared with therapeutic serum concentrations of the compounds in patients, and in vivo LD50 values from animal studies. The in vitro results correlated well with the in vivo data, so the test appears to reflect immunomodulation. Results were reproducible (CV = 20 +/- 5%), and the method could be transferred to another laboratory (r(2) = 0.99). We therefore propose this method for further validation and for use in immunotoxicity testing strategies.
authors.affiliation: European Centre for the Validation of Alternative Methods, Institute for Health & Consumer Protection, European Commission Joint Research Centre, 21020 Ispra (VA), Italy.
authors.forename: Ingrid
authors.initials: I
authors.lastname: Langezaal
authors.forename: Sebastian
authors.initials: S
authors.lastname: Hoffmann
authors.forename: Thomas
authors.initials: T
authors.lastname: Hartung
authors.forename: Sandra
authors.initials: S
authors.lastname: Coecke
mesh_terms: D000276:Adjuvants, Immunologic / Q000633:toxicity*; D000826:Animal Testing Alternatives; D000818:Animals; D001773:Blood Cells / Q000187:drug effects* / Q000378:metabolism; D016207:Cytokines / Q000097:blood*; D003602:Cytotoxicity, Immunologic; D004305:Dose-Response Relationship, Drug; D064420:Drug-Related Side Effects and Adverse Reactions; D004768:Enterotoxins / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D066298:In Vitro Techniques; D007928:Lethal Dose 50; D008070:Lipopolysaccharides / Q000494:pharmacology; D015203:Reproducibility of Results
chemical_list: D000276:Adjuvants, Immunologic; D016207:Cytokines; D004768:Enterotoxins; D008070:Lipopolysaccharides; C031818:enterotoxin B, staphylococcal