GM-CSF与猪瘟病毒E2基因共表达载体的构建及其诱导的免疫应答

为探讨粒细胞—巨噬细胞集落刺激因子(GM-CSF)对猪瘟病毒E2基因核酸疫苗小鼠免疫应答诱导的影响,将CSFVE2基因和GM-CSF片段插入真核表达载体pcDNA3.0内构建pcE2,pcE2-GF及pcGF核酸疫苗.动物试验时,50只雌性供试小鼠随机分为5组,即pcE2,pcE2-GF,pcDNA3.0,pcGF,生理盐水组,每组10只.0,2,4周于小鼠左后肢胫前肌进行肌注免疫,质粒量为每只每次50μg.每次免疫前和末次免疫后2周尾静脉采血收集血清,ELISA检测血清内特异性IgG水平.末次免疫后1周每组随机选3只无菌取脾,MTT法检测淋巴细胞增殖情况.结果表明,与对照组比较,pcE2和pcE2-GF第3次免疫2周后,免疫小鼠IgG抗体水平逐渐增高,淋巴细胞的转化率也明显增高,且pcE2-GF免疫组的IgG抗体水平和淋巴细胞的转化率高于pcE2免疫组.可见,细胞因子GM-CSF可有效提高猪瘟病毒E2基因核酸疫苗的免疫应答.

Immune response induced by DNA vaccine Co-encoding of classical swine fever virus E2 and GM-CSF in mice

To study effect of GM-CSF on immune response of mice elicited by immunization with DNA vaccine encoding of classical swine fever virus E2,CSFVE2 and GM-CSF amplified by PCR.were inserted into pcDNA3.0 to construct:pcE2,pcE2-GF and pcGF.50 female mice were randomly divided into 5 groups,10 each.Group A,B were immunized with pcE2 and pcE2-GF and group B,C and D are controls,immunized with pcDNA3.0,pc GF and physiological saline respectively.At weeks 0,2,4,sera were collected before every immunization and 2 weeks after final immunization.Serum lgG were detected by ELISA.Spleen lymphocyte cells of mice were collected 2 weeks after final immunization.Proliferation activity of spleen lymphocytes was detected by MTT assay.Results showed that compared with control groups,level of IgG in sera increased,and proliferation activity of spleen lymphocytes were increased.FurthermoreI,gG level in sera and proliferation activity of spleen lymphocytes of group B are higher than that of group A.In conclusion,GM-CSF can increase the level of immune response in mice induced by immunization with DNA vaccine encoding of classical swine fever virus E2.