E.tenella河北株pcDNA3.0-IL-2-EtMIC-2真核表达载体的免疫调节作用

Effects of the immunomodulatory of eukaryotic expression vector pcDNA3.0 IL-2-EtMIC-2 of Eimeria tenella of Hebei strain

为了研究柔嫩艾美耳球虫(E.tenella)河北株真核表达载体pcDNA3.0-IL-2-EtMIC-2的免疫调节作用,将240只14日龄雏鸡随机分成8组,第1,2组分别为阳性、阴性对照,在14,21 d,3、4组分别肌注100μg的pcDNA3.0-IL-2、pcDNA3.0-EtMIC-2;5、6、7组分别肌注50,100,150μg的pcDNA3.0-IL-2-EtMIC-2;8组21 d时滴口免疫鸡球虫病四价活疫苗。28 d,阳性对照组和6个试验组每只鸡经口攻毒4×10~4个E.tenella卵囊,阴性对照组每只鸡灌服同体积的生理盐水。结果显示:当pcDNA3.0-IL-2-EtMIC-2真核表达载体的免疫剂量达100,150μg/只时,试验后7,21,28 d IgG的含量、7 d的IgA含量和21,28 d的IFN-γ含量均显著高于阴性对照组、阳性对照组、单独免疫pcDNA3.0-IL-2和pcDNA3.0-EtMIC-2组(P<0.05);试验后28 d,血清中IL-2的含量显著高于阴性和阳性对照组(P<0.05),与单独免疫pcDNA3.0-IL-2和pcDNA3.0-EtMIC-2组相比差异不显著(P>0.05);试验后7,14,21,28 d脾脏IL-2和IFN-γmRNA表达量均显著高于阳性对照组、单独免疫pcDNA3.0-IL-2和pcDNA3.0-EtMIC-2组(P<0.05)。综上所述,IL-2可增强EtMIC-2基因的免疫原性,适量的真核表达载体pcDNA3.0-IL-2-EtMIC-2可显著提高感染柔嫩艾美尔球虫雏鸡的免疫功能。

To study the immunoregulatory effects of eukaryotic expression vector pcDNA3.0-IL-2-EtMIC-2 of E.tenella Hebei strain,240 14-day-old chicks were randomly divided into 8 groups.The first and second groups were positive and negative controls respectively.At 14 and 21 days,groups 3 and 4 were intramuscularly injected with 100μg of pcDNA3.0-IL-2 and pcDNA3.0.-EtMIC-2,5,6,and 7 groups were intramuscularly injected with 50,100,and 150μg of pcDNA3.0-IL-2-EtMIC-2,and 8 groups of mice were immunized with a four-valent live vaccine for oral coccidiosis.On the 28 th day,each chicken in the positive control group and the 6 test groups was challenged with 4×104 E.tenella oocysts by mouth,and each chicken in the negative control group was given the same volume of normal saline.The results showed that when the immunization dose of pcDNA3.0-IL-2-EtMIC-2 eukaryotic expression vector reached 100,150μg/head,the serum of IgG at 7,21,28 d,the content of IgA at 7 d,the content of IFN-γat 21,28 d,were significantly increased compared with the negative control group,positive control group,pcDNA3.0-IL-2 group and pcDNA3.0-EtMIC-2 group(P<0.05),At 28 days after the test,the serum IL-2 content was significantly higher than that of the negative and positive control groups(P<0.05),and the difference was not significant compared with the immunized pcDNA3.0-IL-2 and pcDNA3.0-EtMIC-2 groups alone(P>0.05);At 7,21,28 d,the expressions of IL-2 and IFN-γmRNA in spleen were significantly higher than those in the positive control group and pcDNA3.0-IL-2 and pcDNA3.0-EtMIC-2 groups(P<0.05).In summary,IL-2 can enhance the immunogenicity of EtMIC-2 gene,and the appropriate amount of eukaryotic expression vector pcDNA3.0-IL-2-EtMIC-2 can significantly improve the immune function of chicks infected with E.tenella.