A new colorimetric method for measuring cell-mediated cytotoxicity in dogs.

In order to replace the radioactive 51chromium release assay (CRA), a colorimetric method for the determination of cell-mediated cytotoxicity of natural killer (NK) effector cells of dogs and adherent target cells was developed using the dye Rose Bengal (RB). After a 14 h incubation period of leucocytes isolated from the peripheral blood (PBL) of dogs and a natural killer cell-sensitive canine adenocarcinoma cell line (CTAC), effector and lysed target cells were removed by washing, and the surviving adherent target cells were stained with RB. The optical density (OD) of the remaining target cells was measured in a microspectrophotometer (ELISA reader) and was found to correspond to the number of surviving cells, and thus was inversely correlated to the cytotoxic activity. The RB assay revealed almost identical cytotoxic values when compared with the CRA. In contrast to this assay the RBA is quick and easy to perform, inexpensive and avoids radioactive materials and waste. However, the method is restricted to adherent target cells.

[Two mesenchymal tumor cell lines for the determination of natural killer (NK) cell activity in the peripheral blood of dogs]. / Zwei mesenchymale Tumorzellinien zur Bestimmung der Natürlichen Killer (NK)-Zell-Aktivität im peripheren Blut des Hundes.

The two mesenchymal cell lines K1 and K6 were established from round cell tumours located at the lips of two dogs. Both cell lines were characterized as being of myelomonocytic origin by morphological, cytochemical, immunocytochemical and functional criteria. In the 51chromium release assay (effector/target cell ratio 100/1) the K6-cell line revealed a mean cytotoxic sensitivity of 53.6% and thus showed a susceptibility similar to that of the epithelial CTAC line (57.8%). The K1-cell line exhibited less cytotoxic activity (41%) when incubated for 14 h, but showed results comparable to the K6-cell line, when the incubation time was reduced to 8 h.