Abstract: Pick-off blood with anticoagulant aseptically and the lymphocytes were isolated. Effects of FCS，PBMC，PHA-P of different concentration on promote lymphocytes proliferation were tested by orthogonal test. On the base of the best level combination, different concentration canine or pig TF was investigated to detect its effect on the activation of lymphocytes by MTT essay. The result of experiment demonstrate that the best levels combination of canine lymphocyte, PHA-P, calf blood serum were 2.5×106 unit/ml，6.25μg/ml，10% respectively. Under this optimum cultural condition, the lymphocytes remarkable proliferated between 0.195~25mg/ml of canine and pig TF，but the best stimulate concentration of the canine and pig TF only or it with PHA-P is 1.56mg/ml. Furthermore, it is confirmed that lymphocytes developed by the canine and pig TF single was much better than they cooperated with PHA-P. The results confirm that the same or difficult race TF’s effect was obvious and provides a basis for further application on the difficult race TF preventing and curing canine’s virus disease.