黄芪多糖与复方黄芪多糖对猪瘟疫苗免疫效果影响的研究
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摘要
研究目的:本实验的主要目的在于探索黄芪多糖(Astragalus polysaccharide,APS)和复方黄芪多糖(Compound of astragalus polysaccharides,Co-APS)注射液作为猪瘟活疫苗稀释液以及黄芪多糖和复方黄芪多糖作为猪瘟活疫苗的佐剂或者现有猪瘟疫苗专用稀释液的改良剂。试验方法:以往的研究表明黄芪多糖具有抗病毒,甚至是杀灭病毒的作用,所以本实验通过体外实验,兔体、猪体两种猪瘟疫苗实验动物的体内实验三部分的实验,观察黄芪多糖对猪瘟活疫苗的影响以及它在动物体内对免疫效果的影响。首先,体外实验运用MTT法测定细胞活性和荧光RT-PCR法测定细胞液中猪瘟病毒含量,观察黄芪多糖和复方黄芪多糖注射液对猪睾丸细胞系(Swine testicular cells,ST)细胞活性的影响以及它对猪瘟疫苗弱毒株在猪睾丸细胞系上生长的影响。其次,运用淋巴细胞转化试验、酶联免疫吸附实验,观察黄芪多糖注射液对兔体外周血液中淋巴细胞活性和血清中的淋巴因子含量以及猪瘟抗体水平的影响。最后,进行猪体实验,运用淋巴细胞转化试验、酶联免疫吸附实验,观察黄芪多糖注射液对兔体外周血液中淋巴细胞活性和血清中的淋巴因子含量、猪瘟抗体水平以及免疫球蛋白含量的影响。通过这三个部分的实验来探讨黄芪多糖注射液作为猪瘟活疫苗稀释液以及黄芪多糖作为猪瘟活疫苗的佐剂或者现有猪瘟疫苗专用稀释液的改良剂的可行性。实验结果:体外实验表明,黄芪多糖和复方黄芪多糖对猪睾丸细胞系细胞具有一定的安全浓度范围,在这个安全浓度范围中:高浓度的黄芪多糖具有提高猪睾丸细胞系细胞活性,抑制猪瘟疫苗弱毒株在猪睾丸细胞系上复制的作用;相对而言,适宜浓度的黄芪多糖具有提高猪睾丸细胞系细胞活性作用,同时具有促进猪瘟疫苗弱毒在猪睾丸细胞系上生长的效果。兔体实验表明,黄芪多糖具有提高兔体外周血液中淋巴细胞细胞活性,提升血清中多种淋巴细胞因子含量以及猪瘟抗体水平的作用。猪体实验表明,黄芪多糖具有提高猪体外周血液中淋巴细胞细胞活性,提升血清中多种淋巴细胞因子含量、猪瘟抗体水平以及免疫球蛋白含量的作用。结论:通过体外、兔体、猪体体内三部分实验证明,黄芪多糖注射液完全可以作为猪瘟活疫苗稀释液以及黄芪多糖作为猪瘟活疫苗的佐剂或者现有猪瘟疫苗专用稀释液的改良剂。
Objective: To explore the possibility of astragalus polysaccharide and compound of astragalus polysaccharides injection using to adjuvant and diluents or its ameliorant of swine fever active vaccine. Method: For the antiviral and killing virus activities, the in vitro and in vivo (by animal modal of rabbit and pig) experiments were taken out to observe the vaccination effect of astragalus polysaccharide and compound of astragalus polysaccharides injection to swine fever active vaccine. First, the activity of swine testicular cells and the content of swine fever virus of swine testicular cells impacted by astragalus polysaccharide and compound of astragalus polysaccharides injection were determined by method of MTT and fluorescence RT-PCR respectively. Then, lymphocyte proliferation assay and enzyme linked immunosrodent assay were taken out to survey the vaccination effect of astragalus polysaccharides injection by determining lymphocyte proliferation activity, the content of lymphokine in serum, swine fever antibody and immune globulin. Results: in vitro experiment, astragalus polysaccharide and compound of astragalus polysaccharides injection had a safety concentration range, and astragalus polysaccharides injection in the higher concentration could promote the activity of swine testicular cells and restrain the swine fever vaccine low virulent strain duplicating on swine testicular cells. And astragalus polysaccharides injection in proper concentration could promote the activity of swine testicular cells and the swine fever vaccine low virulent strain duplicating on swine testicular cells. In vivo experiment of rabbit, astragalus polysaccharides had the effect of increasing lymphocyte proliferation, the content of lymphokine in serum and swine fever antibody. In vivo experiment of pig, astragalus polysaccharides had the effect of increasing lymphocyte activty, the content of lymphokine in serum, the level of swine fever antibody and immune globulin. Conclusion, the in vitro and in vivo (by animal modal of rabbit and pig) experiments proved that astragalus polysaccharide injection could be used as adjuvant and diluents or its ameliorant of swine fever active vaccine.
Objective: To explore the possibility of astragalus polysaccharide and compound of astragalus polysaccharides injection using to adjuvant and diluents or its ameliorant of swine fever active vaccine. Method: For the antiviral and killing virus activities, the in vitro and in vivo (by animal modal of rabbit and pig) experiments were taken out to observe the vaccination effect of astragalus polysaccharide and compound of astragalus polysaccharides injection to swine fever active vaccine. First, the activity of swine testicular cells and the content of swine fever virus of swine testicular cells impacted by astragalus polysaccharide and compound of astragalus polysaccharides injection were determined by method of MTT and fluorescence RT-PCR respectively. Then, lymphocyte proliferation assay and enzyme linked immunosrodent assay were taken out to survey the vaccination effect of astragalus polysaccharides injection by determining lymphocyte proliferation activity, the content of lymphokine in serum, swine fever antibody and immune globulin. Results: in vitro experiment, astragalus polysaccharide and compound of astragalus polysaccharides injection had a safety concentration range, and astragalus polysaccharides injection in the higher concentration could promote the activity of swine testicular cells and restrain the swine fever vaccine low virulent strain duplicating on swine testicular cells. And astragalus polysaccharides injection in proper concentration could promote the activity of swine testicular cells and the swine fever vaccine low virulent strain duplicating on swine testicular cells. In vivo experiment of rabbit, astragalus polysaccharides had the effect of increasing lymphocyte proliferation, the content of lymphokine in serum and swine fever antibody. In vivo experiment of pig, astragalus polysaccharides had the effect of increasing lymphocyte activty, the content of lymphokine in serum, the level of swine fever antibody and immune globulin. Conclusion, the in vitro and in vivo (by animal modal of rabbit and pig) experiments proved that astragalus polysaccharide injection could be used as adjuvant and diluents or its ameliorant of swine fever active vaccine.
引文
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