Abstract
Blastogenesis of canine peripheral blood lymphocytes (PBL) against phytohemagglutinin (PHA) was measured by the glucose consumption test (GCT). Two hundred thousand lymphocytes isolated from a dog were placed in a flat-bottomed microplate in DM-160 medium supplemented with 15% normal canine serum (incubated at 56°C for 30 minutes) and 20μg/ml PHA-P (final concentration). The plate was sealed tightly and incubated for 96 hr at 38°C. The mean of SR value on 34 clinically healthy dogs was 73.8±20.0% under those culture conditions and there was a high correlation (r=0.79, P<0.01) of results obtained between this test and the fluorometric microassay of DNA in mononuclear cell using ethidium bromide. The blastogenesis of PBL was inhibited significantly (P<0.05) by the treatment with mitomycin C. The SR value of lymphocytes of which isolated from six dogs with experimental lobectomy was decreased from 49.7±22.9% to 16.3±15.0% after operation. GCT was able to reveal a statistical depression (P<0.05) of lymphocyte blastogenesis after operation. In brief, this test could detect efficiently a depression of lymphocyte blastogenesis in vitro and in vivo. In conclusion, this GCT was regarded as a new effective method for measurement of blastogenesis of canine PBL.
