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鲤I-IFN-γ作为锦鲤疱疹病毒核酸疫苗免疫佐剂的效果研究
于慧,周井祥
0
(吉林农业大学动物科学技术学院)
摘要:
为了研究鲤Ⅰ-IFN-γ的免疫效果,根据鲤Ⅰ-IFN-γ的mRNA基因序列(GenBank),构建真核表达质粒pEGFP-N1-Ⅰ-IFN-γ,重组表达质粒与不同剂量的转染试剂EndoFectionTM Max混合后转染鲤鱼肌肉细胞,通过荧光倒置显微镜观察,重组表达质粒pEGFP-N1-Ⅰ-IFN-γ在细胞中成功表达。以多种免疫剂量的重组表达质粒与pIRES-ORF81混合对鲤鱼进行肌肉注射,每周免疫一次,共免疫三次,免疫前采血,收集血清,通过酶联免疫吸附试验(ELISA)检测抗体水平。结果表明:于第三次免疫后两周,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,抗体水平明显升高,但差异不显著(P>0.05)。鲤Ⅰ-IFN-γ作为锦鲤疱疹病毒的免疫佐剂具有一定的免疫效果。
关键词:  鲤I型γ干扰素  锦鲤疱疹病毒  核酸疫苗pIRES-ORF81  免疫效果
DOI:
投稿时间:2016-07-04修订日期:2016-08-05
基金项目:公益性行业专项-锦鲤疱疹病毒病核酸疫苗的研制及初步应用(106037-2016-2018);国家现代农业产业体系项目(CARS-46-29)
Immune Effect of Carp I-IFN-γ as Koi Herpesvirus Nucleic Acid Vaccine Adjuvants
(JiLin Agricultural University)
Abstract:
Study on the carp Ⅰ-IFN-γ immune effect, on the basis ofⅠ-IFN mRNA gene sequence Genbank login, The eukaryotic recombinant plasmid pEGFP-N1-Ⅰ-IFN were constructed.Mixing recombinant expression plasmid and transfection reagents in several immune dose transfected carp muscle cells. The expression of recombinant expression plasmid pEGFP-N1-Ⅰ-IFN-γ in carp muscle cells were observed by fluorescent inverted microscope.By using the mixture of recombinant expression plasmidin several immune dose and pIRES - ORF81 injected Carp′s intramuscular and immunized once a week, three times in total. Blood was sampling before injection and serum was collected to detect antibody levels by enzyme-linked immunosorbent assay(ELISA). Two weeks after the third immunization,experiment result showed that antibody titer reach the highest level. Compared with pIRES - ORF81alone immune,antibody level of combined immunization increased obviously, but no significant difference was observed between them(P>0.05). Show that carp Ⅰ - IFN -γ have certain immune effectas koi herpesvirus immune adjuvant.
Key words:  Ⅰ-IFN-γ  koi herpesvirus  nucleic acid vaccine pIRES- ORF81  immune effect

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