猪轮状病毒与传染性胃肠炎病毒核酸免疫研究

将昆明鼠随机分为A,B,C,D4组,各组分别采用含有猪轮状病毒(RV)JL94株VP7基因的重组真核表达质粒pcDNA-VP7,含有猪传染性胃肠炎病毒(TGEV)TH98株N基因的重组真核表达质粒pcDNA-N,pcD-NA-VP7和pcDNA-N、真核表达载体pcDNA3.1(+),肌肉注射3次,每次间隔2周。首次注射前后定期采血,检测血清抗体和外周血淋巴细胞中CD4+,CD8+T细胞数量的变化。A,C组小鼠血清在首免后第14天即可检出针对RVVP7的阳性抗体(P/N≥2.0)。B组小鼠血清在首免后第39天检出针对TGEVN蛋白的阳性抗体(P/N≥2.0)。A,B,C组小鼠在首免后外周血CD4+、CD8+T细胞数量与D组小鼠相比,在不同时间有显著差异(P<0.05),并明显高于D组小鼠。说明猪轮状病毒与传染性胃肠炎病毒核酸免疫后能诱发机体免疫反应。

Study on the nucleic acid immunization of procine rotavirus and transmissible gastroenteritis virus

The Kunming mice used for nucleic acid immunization were randomly divided into four groups(A,B,C and D).Group A was injected intramuscularly with the recombinant plasmid,pcDNA-VP7,which contained VP7 gene of the native JL94 isolate of porcine rotavirus(RV),group B with the recombinant plasmid,pcDNA-N,which contained N gene of the TH98 isolate of transmissible gastroenteritis virus(TGEV),group C with the combinative pcDNA-VP7 and pcDNA-N;group D with empty eukaryotic expression plasmid pcDNA3.1(+).The mice in each group were injected three times totally,and the interval of every time was two weeks.Blood samples collected at regular intervals from mice before and after primary injection,were used for detecting specific antibody levels in serum and the differences in number of CD4+,CD8+ T cells in peripheral blood lymphocytes. The results showed that the antibodies to RV could be detected in the serum of group A and C mice from 14 days post-immunization(PI),the antibodies to TGEV N protein could be detected in the serum of group B mice at 39 days of PI.Comparing with group D,there were significant differences(P<0.05) in number of CD4+,CD8+ T cells in peripheral blood lymphocytes of group A,B,C at different time,furthermore,the number of CD4+,CD8+ T cells of each group were higher than group D.This elucidated that the immune response of mice could be induced after immunized with porcine rotavirus and transmissible gastroenteritis virus nucleic acid.