鲤HSP90作为锦鲤疱疹病毒核酸疫苗免疫佐剂的效果研究

为研究鲤热休克蛋白90(HSP90)的免疫效果,根据鲤HSP90 mRNA的基因序列(GenBank),构建真核表达质粒pEGFP-C1-HSP90,并将pEGFP-C1-HSP90转染至鲤脑细胞(CCB)中,荧光倒置显微镜下观察显示,重组质粒pEGFP-C1-HSP90在CCB中成功表达;以不同免疫剂量的重组表达质粒与pIRESORF81联合免疫鲤,通过间接酶联免疫吸附试验(ELISA)检测免疫前后抗体水平。结果表明:注射重组质粒的血清中能够检测到KHV特异性抗体,其抗体水平随着免疫次数的增加而增加,第三次免疫后两周,抗体水平达到最高;联合免疫与单独免疫核酸疫苗pIRES-ORF81相比,抗体水平明显增加,但无显著性差异(P0.05)。研究表明,鲤HSP90作为锦鲤疱疹病毒的免疫佐剂具有一定的目标效果。

Immune effect of carp HSP90 as an adjuvant of koi herpesvirus nucleic acid vaccine

The eukaryotic recombinant plasmid pEGFP-C1-HSP90 as an adjuvant was constructed on the basis of carp HPS90 mRNA gene sequence GenBank login,and then transfected into brain cells of common carp Cyprinus carpio (CCB),being observed by fluorescent inverted microscope,in order to study the immune effect of HSP90. The antibody levels were detected in the common carp injected intramuscularly with the mixture of recombinant ex-pression plasmid and pIRES-ORF81 at several immune doses by indirect enzyme-linked immunosorbent assay (ELISA). The results showed that serum injection of recombinant plasmid KHV was detected in the specific anti-body in the common carp,with increase in the antibody level with the increase in immunization frequency,the peak in third and two weeks after immunization. There was higher antibody level in combined immunization than that in pIRES-ORF81 immunization alone,without significant difference(P>0.05),indicating that the recombinant plas-mid pEGFP-C1-HSP90 as adjuvant of koi herpes virus has certain immune effects.