Techniques for assessing canine mononuclear phagocyte function as part of an immunotoxicologic evaluation

Abstract

Although the dog is used extensively as a model for toxicologic investigation, few practical techniques for assessing immunotoxicity in this species have been reported. We have adapted techniques for assessing peripheral blood monocyte (PBM) function in the dog. Methods for isolating PBM were compared; a technique employing hypertonic conditioning followed by density gradient centrifugation, which resulted in > 90% purity and a greater yield of PBM than that obtained with other previously reported methods is described, along with methods for measuring antibody-dependent cellular cytotoxicity (ADCC) and the production of prostaglandin E 2 (PGE 2) and superoxide anion (O 2 −). An assay for the measurement of interleukin-1 (IL-1), based on its ability to induce interleukin-2 (IL-2) production by the EL-4 mouse thymoma line in the presence of the calcium ionophore A23187, was examined. Together, these assays provide the tools to better define drug or chemical effects on the mononuclear phagocyte system (MPS) of this important animal model.