Studies On The Cellular And Humoral Immunities In BALB/c Mice Immunized With VP3 Gene Vaccine Of Goose Parvovirus

In this study, BALB/c mice were immunized with pcDNA-GPV-VP3 DNA vaccine by gene gun bombardment( 1μg, 3μg, 6μg)and intramuscular injection( 50μg, 100μg, 200μg), live attenuated vaccine of Sichuan isolate of GPV, pcDNA 3.1 (+) and 0.09%NaC1 were controls. Collected the peripheral blood samples at 3d,7d,14 d,21d,28d,35d,49d,63d,77d,105d,133d,161d,189dand217 dafter immunized. T lymphocyte proliferation test (MTT), Fluorescence Activated Cell Sorter (FACS) and indirect ELISA method were employed respectively to detect cellular and humoral immune responses of BALB/c mice immuned with pcDNA- GPV-VP3 DNA vaccine. Results:1. Peripheral T-lymphocytes responses:â‘ Peripheral T-lymphocytes responses to ConA of the experiment groups (except im 200μg group) and the control groups were significantly different (P＜0.01) or different (P＜0.05). The values were increased from 14 d, chalked to the highest point at 28 d and then degraded after immunization. Otherwise the pcDNA 3.1(+) and 0.09%NaC1 control groups were significantly differrent (P＜0.01) at 7-63 d after immunized.â‘¡Peripheral T lymphocytes proliferation of 6μg group vaccine by gene gun bombardment induced the best response, 3μg group was the better one, 1μg group was the wost one and it showed some dose-dependent character.â‘¢Peripheral T- lympho cytes responses of 100μg to ConA vaccined by intramuscular injection group and 50, 200μg groups were significantly different (P＜0.01). And the response of 200μg group was similar with 0.09%NaC1 control group.â‘£Compared the groups which were immunized by gene gun bombardment and intramuscular injection, Peripheral T-lymphocytes responses to ConA were no different.⑤Compared the pcDNA-GPV-VP3 DNA vaccine groups with live attenuated vaccine control group, Peripheral T-lymphocytes responses of 3, 6, 50, 100μg groups and live attenuated vaccine control group were significantly different (P＜0.01) or different (P＜0.05) at 28d,7-105 d,28d,21-35 d.2. The number of CD4⁺ and CD8⁺ T-lymphocytes:â‘ The number of CD4⁺ and CD8⁺ T-lymphocytes after immunized the pcDNA-GPV-VP3 DNA vaccine, the experiment groups and pcDNA 3.1(+), 0.09%NaC1 control groups were significantly different (P＜0.01) or different (P＜0.05) in different times. The experiment groups chalked to the highest value at 28 dâ‘¡After vaccined by gene gun bombardment, the 6μg group induced much more number of CD4⁺ T-lymphocytes and no different between gene gun bombardment groups of CD8⁺ T-lymphocytesâ‘¢After immunized by intramuscular injection, the 50 lag group induced much more number of CD4^, CD8 T-lymphocytes than 100,200μg groups.â‘£Compared the groups which were immunized by gene gun bombardment and intra- muscular injection, 6μg groups and 50μg group indued the number of CD4⁺, CD8⁺T T-lymphocytes higher than other groups and no defferent between the other groups.⑤Compared the pcDNA-GPV-VP3 DNA vaccine groups with live attenuated vaccine contro1 group, 3, 6μg groups and 50μg group indued the number of CD4⁺, CD8⁺T T-lymphocytes higher than live attenuated vaccine control group.3. The changes of anti-GPV IgG in the peripheral blood:â‘ The OD₄₅₀ values of anti-GPV IgG induced by the pcDNA-GPV-VP3 DNA vaccine, the experiment groups and pcDNA 3.1(+), 0.09%NaC1 control groups were significantly different (P＜0.01) or different (P＜0.05) in different times. The values were increased from 14 d, chalked to the highest point at 28 d and then degraded after immunization. But the titres were low.â‘¡The OD₄₅₀ values ofanti-GPV IgG induced by 6μg group was the highest one, 3μg group was the better one, 1μg group was the wost one and it showed some dose-dependent character.â‘¢After immunized by intramuscular injection, the OD450 value of anti-GPV IgG induced by 50μg group was the highest oneâ‘£Compared the groups which were immunized by gene gun bombardment and intra- muscular injection, they were no different.⑤Compared the pcDNA-GPV-VP3 DNA vaccine groups with live attenuated vaccine control group, the OD₄₅₀ value of anti-GPV IgG induced by 6, 50 ,100μg groups and live attenuated vaccine control group significantly different (P＜0.01) or different (P＜0.05) at 21-35 d and 21 d.