Effect of cell mediated immunity regulation of duck enhanced by duck IFN-α eukaryon expression plasmid and inoculated with DPV attenuated vaccine by gene-gun

Abstract

In order to study the effect of cell mediated immunity regulation of duck IFN-α eukaryon expression plasmid (pcDNA-SDIFN-α) on duck plague virus (DPV) attenuated vaccine in ducks, pcDNA-SDIFN-α was administered to 28-day-old ducks at doses of 1, 3 and 6 μg per duck, respectively, by gene-gun. PBS and empty vector pcDNA were used as control. Fifteen days later, all ducks were injected with DPV attenuated vaccine and blood samples were collected at 3, 7, 14, 21, 28, 35, 49, 63 and 84 days after injection. T-lymphocyte proliferation tests (MTT) were used to detect the T-lymphocyte proliferation in the peripheral blood (PBL) of ducks. Blood samples collected at 7, 14, 21, 28, 35 and 49 days after injection were detected by fluorescence-activated cell sorter (FACS) for recording the number of CD3+ T-lymphocytes of ducks. Results were as follows: (1) Reaction of T-lymphocytes in PBL to ConA (OD value) of ducks treated with pcDNA-SDIFN-α was higher than that of PBS and pcDNA control groups in 3–84 days. There were highly significant differences between the 1 μg per duck group and the two control groups in 3–84 days (P ⩽ (0.01), between the 3 μg per duck group and the two control groups in 3–84 days (P ⩽ 0.01, P ⩽ 0.05), and between the 6 μg per duck group and the two control groups in 7–49 days (P ⩽ 0.01, P ⩽ 0.05). The significant difference was also present between the groups of 1, 3 and 6 μg per duck in 3–35 days (P ⩽ 0.05). However, there was no significant difference between the 3 and 6 μg per duck groups (P ⩾ 0.05). The pcDNA control group was higher than PBS control group, but no difference was detected (P ⩾ 0.05). (2) Change of the number of CD3+ T-lymphocytes in ducks administered with different doses of pcDNA-SDIFN-α was higher than that of PBS and pcDNA control groups in 7–49 days. The change in the 1 μg per duck group was significantly higher than that in PBS and pcDNA control groups in 14–49 days (P ⩽ 0.01). There were significant differences between the 3 μg per duck group and the two control groups in 21–49 days (P ⩽ 0.01, P ⩽ 0.05) and between the 6 μg per duck group and the two control groups in 7–49 days (P ⩽ 0.01, P ⩽ 0.05). However, no significant differences among the groups of 1, 3, and 6 μg per duck groups (P ⩾ 0.05) and between the two control groups (P ⩾ 0.05) were found. The results indicated that pcDNA-SDIFN-α administered 15 days before injection of DPV-attenuated vaccine could significantly enhance cellular immunity induced by DPV-attenuated vaccine. pcDNA-SDIFN-α is an excellent DPV-attenuated vaccine molecular adjuvant and the best result can be obtained with the dose of 1 μg per duck of pcDNA-SDIFN-α inoculated by gene-gun.