title: Validation of an eight parameter immunophenotyping panel in adult canines for assessment of immunotoxicity.
abstract: Analysis of peripheral blood leukocyte populations by flow cytometry in adult beagles is a critical component of immunotoxicity assessment in regulated pre-clinical toxicology studies. In this study, data is presented utilizing a single panel, six-color method to simultaneously enumerate absolute cell counts and determine the relative percentage of leukocytes. A GLP validation was performed to determine intra- and inter-assay variance, inter-instrument variance, and pre- and post-fixation stability for the target populations. The results demonstrated all samples met acceptance criteria, CV values less than 25%, for all precision and stability intervals assessed. The intra and inter-assay data demonstrated the single panel method generated acceptable precision. Furthermore, stability results indicated whole blood samples and processed samples may be stored without a statistically significant difference in the data compared to samples immediately processed and analyzed after blood collection. This assay will provide researchers a more precise and efficient tool to evaluate the immunotoxic effects of a test article on canine peripheral blood leukocytes during pre-clinical drug testing.
authors.affiliation: Department of Cell and Molecular Biology, MPI Research, 54943 North Main Street, Mattawan, MI 49009, USA. Brandon.Zeigler@mpiresearch.com
authors.forename: Brandon M
authors.initials: BM
authors.lastname: Zeigler
authors.forename: Yvonne
authors.initials: Y
authors.lastname: Boyle-Holmes
authors.forename: Deanna
authors.initials: D
authors.lastname: Falzone
authors.forename: John
authors.initials: J
authors.lastname: Farmer
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000235:genetics / Q000378:metabolism*; D004285:Dogs / Q000276:immunology* / Q000378:metabolism*; D005260:Female; D005434:Flow Cytometry / Q000662:veterinary; D005786:Gene Expression Regulation / Q000276:immunology*; D007962:Leukocytes / Q000378:metabolism*; D008297:Male; D015203:Reproducibility of Results
chemical_list: D015703:Antigens, CD