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Study on Immunoregulation Effect of Duck IFN-α Eukaryon Expression Plasmid to Ducks Immunized with DP Attenuated Vaccine

Author: ChengZhiPing
Tutor: WangMingShu;ChengAnChun
School: Sichuan Agricultural University
Course: Preventive Veterinary Medicine
Keywords: Gene gun Duck IFN - a eukaryotic expression plasmid The duck plague attenuated vaccine Cell-mediated immunity Humoral immunity Molecular adjuvant
CLC: S858.32
Type: Master's thesis
Year: 2007
Downloads: 59
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Abstract


Duck eukaryotic expression plasmid, alpha-interferon (pcDNA the-SDIFN-alpha), three doses of each 1,3,6 g Bombardment 50,100,200 μg per three-dose intramuscular injection method were immunized with the 28-day-old duck, and 15 d after inoculation of all duck duck plague (DP) attenuated vaccine, at the same time set the empty plasmid pcDNA3.1 () control group, DP the attenuated control group and blank control group, after inoculation 3,7 14,21,28,35,49,63,84,105 d mining anticoagulant lymphocyte proliferation test (MTT method) Determination of duck peripheral blood T lymphocyte transformation 7,14,21,28,35,49; d adopt anti-clotting measured by flow cytometry (FACS) CD * T lymphocytes dynamic changes; 7th, 14th, 21st, 28th, 35th, 49,63,84,105,126,154,182 d mining coagulation indirect ELISA and neutralization test for detection of serum antibody levels; 12 h, 24 h, 3 d, 7 d, 14 d, 21 d, 28 d, 35 d, 49 d, 63 d, 84 d adopt anti-clotting quantitative real-time PCR detection of blood in the DPV-DNA content. For the following results: 1. Ability of the reaction of T lymphocytes to ConA (OD value) the the pcDNA-SDIFN-alpha to a different dose, different ways immunization Ap, re-immunization DP attenuated in 3 d, the reaction ability of T lymphocytes to ConA was significantly higher than that attenuated control group, empty vector control group and blank control group, 28 d peak. The experimental group is extremely significant with (P ≤ 0.01) or significant (P ≤ 0.05) higher than the the Attenuated control group, vector control and blank control group; vector control and attenuated the control group significantly (P ≤ 0.05) higher than the control Group; vector control attenuated slightly higher than the control group, but no significant difference (P ≥ 0.05). 2. CD 3 the number of lymphocytes in the first 7 d the duck peripheral blood CDs 3 T lymphocytes increased significantly , the experimental group duck peripheral blood CD 3 T lymphocytes were significantly (P ≤ 0.01) or significant (P ≤ 0.05) higher than the attenuated control group, empty vector control group and the control group; vector control and attenuated control group significantly (P ≤ 0.05) or highly significant (P ≤ 0.01) higher than the control group; empty vector control group slightly higher than the attenuated control group but no significant difference (P ≥ 0.05). 3. Peripheral blood antibody IgG levels (OD) in the first 7 d, in addition to the control group, all groups duck serum antibody levels increased rapidly, reached a peak in 28 days when. The 1,3,6,200 microg the immunohistochemistry duck serum antibody IgG were significantly (P <0.05) or highly significant (P ≤ 0.01) higher than the the attenuated control group, empty vector control group and blank control group; 50 and 100 ug immunohistochemistry slightly higher Attenuated control group and empty vector control group, but no significant difference (P ≥ 0.05); attenuated control group and empty vector control group level of Ap serum antibody no significant difference (P ≥ 0.05), but a very significant significantly (P ≤ 0.01) or significantly (P ≤ 0.05) higher than the control group. 4. Peripheral blood and antibody titer in the experimental group, attenuated control group and empty vector control group duck duck plague specific antibody, the antibody titer in 28 days when peak. The 1,3,6,200 μg immunohistochemical antibody neutralization titer higher than 50,100 μg immunohistochemistry, the the attenuated control group and empty vector control group antibody titer. 5. DNA content of peripheral blood DPV-in addition to the control group, all groups duck peripheral blood of DPV-DNA in 12 h can be detected to 12 h-49 days was significantly (P ≤ 0.01) higher than the control group, DPV -DNA copy number in order of magnitude of substantially 3.5-4.2. Experimental group, DPV-DNA in the duck peripheral blood of the the attenuated control group and the empty vector control group showed no significant difference (P ≥ 0.05). In summary, the pcDNA-SDIFN-alpha 15 d immune in advance to make DP attenuated vaccine-induced T lymphocyte transformation ability, the CD 3 T lymphocytes increase in the number of serum antibody IgG levels and neutralizing antibody titer rise is a good enhanced DP attenuated vaccine molecular adjuvant cellular and humoral immune the pcDNA-SDIFN-alpha intramuscular injection of 200 microg / only bombardment 1μg / only the best. the the pcDNA-SDIFN-alpha by different ways, different doses of immune duck DPV proliferation in the peripheral blood of ducks. Different ways the immune pcDNA-SDIFN-alpha DP attenuated vaccines stimulate the cellular immune response, Bombardment better than the intramuscular injection method; while in the humoral immune response, intramuscular injection method is better than the Bombardment.

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Livestock, poultry, wildlife diseases > Poultry > Duck
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