Evaluation of immune effect of ESAT6-Fc DNA vaccine against Mycobacterium tuberculosis
10.13200/j.cnki.cjb.003972
- VernacularTitle:结核分枝杆菌ESAT6-Fc DNA疫苗的免疫效应评价
- Author:
YAO Si
- Publication Type:Journal Article
- From:
Chinese Journal of Biologicals
2023;36(8):897-901
- CountryChina
- Language:Chinese
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Abstract:
Objective To evaluate the immune effect of ESAT6-Fc DNA vaccine against Mycobacterium tuberculosis(M.tb),so as to lay a foundation of the next protective test of the vaccine against M.tb infection.Methods The female C57BL/6J mice were randomly divided into two groups with 5 mice in each group,which were injected with the recombinant plasmid pcD-ESAT6-Fc containing HSV2gD signal peptide gene,Rv3875 gene and mouse Fc gene(1 116 bp nucleotides)as well as PBS intramuscularly through thigh,respectively,with the dosage of 50 μg/leg,100 μg in total. After the first immunization,immunization was boosted once a week for three times continuously. Blood samples were collected 14,28,42 d after the first immunization,and the titers of IgG,IgG1 and IgG2a in serum were detected by indirect ELISA;Serum and spleen samples were collected 42 d after the first immunization,spleen cells were isolated and detected for the ratio of CD4~+ and CD8~+T cells secreting IFNγ by flow cytometry,and the contents of IFNγ and IL-2 in serum were determined by ELISA.Results IgG,IgG1 and IgG2a specific antibodies were detected in the serum of mice immunized with the recombinant plasmid 14 d after the first immunization,and the titers continued to increase;42 d after the first immunization,The ratio of CD3~+CD4~+IFNγ~+ and CD3~+CD8~+IFNγ~+ cells and the contents of IFNγ and IL-2 in the recombinant plasmid immunized group were significantly higher than those in the PBS group(t = 12. 43,13. 35,7. 04 and 6. 65,respectively,each P <0. 001).Conclusion The recombinant plasmid pcD-ESAT6-Fc can effectively induce the humoral and cellular immune responses in mice,which lays an experimental foundation for the further development of ESAT6-Fc DNA vaccine.
关键词(KeyWords)： 结核分枝杆菌;Rv3875基因;真核表达载体;体液免疫;细胞免疫;DNA疫苗
Mycobacterium tuberculosis(M.tb);Rv3875gene;Eukaryotic expression vector;Humoral immunity;Cellular immunity;DNA vaccine
