Abstract:

BACKGROUND: The establishment of an effective assessment model for monitoring post-transplant recipient's immune status is the difficulty in current researches.
OBJECTIVE: To establish an in vivo method of cytotoxicity assay in rabbits; to verify whether the in vivo cytotoxicity assay can detect the immune status of the recipient rabbits.
METHODS: New Zealand female-male rabbit skin graft models were established; the untransplanted female and male rabbits served as control. The spleens of both female and male rabbits were removed to prepare single cell suspension; and then live cell dye hydroxyfluorescein diacetate succinimidyl ester of different concentrations were added to stain and prepare for the 1:1 mixed cell suspension. Peripheral blood samples of the male rabbits were collected at 1, 2, 3 and 8 hours after the infusion of the mixed cell suspension. Flow cytometry was used to detect the ratio of the two kinds of fluorescytes in the peripheral blood; the killing rate of specific cells in the donors was calculated.
RESULTS AND CONCLUSION: The female-male rabbit model of skin graft rejection was established in the 2nd week after skin transplantaion. The ratio changes in the peripheral blood of model group and control group were different after the infusion of mixed cell suspension; the killing rate in the male rabbits of the model group was significantly higher than that of the control group at 1, 2, 4 and 8 hours after infusion (P < 0.01). These findings indicate that cytotoxicity assay in vivo can be used to monitor immune state; it can directly reflect the specific immune environment in vivo of the skin graft models and the intensity of the immune attack that the graft suffered.