The comparison of specific antibodies levels in the BALB/c mice immunized with the pcDNA3.1 recombinant plasmids of NS1 gene partial sequence from different dengue type 2 virus strains
10.3760/cma.j.issn.0254-5101.2011.04.016
- VernacularTitle:两株登革2型病毒NS1基因真核重组质粒免疫BALB/c小鼠产生特异性抗体水平的比较
- Author:
Lijuan REN
;
Li ZUO
;
Haidong ZHU
;
Fangfang HU
- Publication Type:Journal Article
- Keywords:
Dengue type 2 virus;
NS1 gene;
Recombinant plasmid;
BALB/c mice;
Humoral immune response
- From:
Chinese Journal of Microbiology and Immunology
2011;31(4):350-355
- CountryChina
- Language:Chinese
-
Abstract:
Objective To compare the humoral immune response of BALB/c mice immunized by recombinant plasmids PeDNA3.1-M-NS1 and pcDNA3.1-N-NS1.Methods Dengue type 2 virus(DENV2)NS1 gene were constructed two partial sequences(1-413 bp)of the pcDNA3.1 eukaryotic plasmids and pET28a(+)plasmid for prokaryotic expression,identification,purification and quantification.The BALB/c mice were immunized by pcDNA3.1-M-NS1,pcDNA3.1-N-NS1 recombinant plasmids with adjuvant.Each animal received a primary inoculation and two boosts at 1-week intervals.Then the blood samples of BALB/c mice were collected from different experiment groups at day 7,14,28 and 56,respectively after first immunization.The specific IgM/IgG antibodies for NS1 protein in serum were confirmed by indirect ELISA.And then the activities of the specific protective antibody were determined by cytopathic effect inhibition(CPEI).Results Construction of the pET28a(+)-NS1 m/pET28a(+)-NS1n prokaryotic expression plasmid,SDS-PAGE analysis showed that,NS1 gene partial sequence was expressed,both the relative molecular weight of about 22.3×103:Western blot showed that the protein can bind anti-His tag monoclonal antibody;byNi affinity chromatographywith apurity of 92% protein,on the C6/36 cell toxicity,and can be used ELASA detection.The results showed that the levels of specific IgM/IgG antibody and neutralizing antibody activities were increased in pcDNA3.1-M-NS1 booster immunization group than other groups.The result had been observed longer duration of antibody level in peDNA3.1-M-NS1 booster immunization group.Conclusion Humoral immune response were significantly different between pcDNA3.1-M-NS1 and pcDNA3.1-N-NS1 recombinant plasmid immunized mice groups.
