Flow cytometoric analysis of chicken NK activity and its use on the effect of restraint stress

Immune system is organized by the influence of both neural and endocrine systems. NK activity plays an important role in the innate immunity. In this study, we observed the effects of restraint stress on chicken peripheral blood NK activity. Viability of FITC-labeled RP9 was measured with PI after treatment with the effector cells. Chicken peripheral blood CD8alpha(+) cells expressed strong cytotoxic activity, in contrast to thrombocytes, while peripheral blood CD3(+) CD8alpha(+) cells and CD4(+) cells had little cytotoxic activity. Con A supernatant enhanced the cytotoxic activity of CD8alpha(+) cells. Therefore, it is considered that these cytotoxic activities measured by flow cytometory (FCM) analysis are NK activity. When chickens were exposed to restraint stress, the levels of serum corticosterone increased transiently over a short period of time while the NK activity decreased. The decreased NK activity, however, did not recover to the intact levels for a long time, even once the serum corticosterone levels had recovered. These data indicate that chicken NK activity is able to be measured by flow cytometric analysis and that restraint stress causes severe damage to the chicken NK activity.