Study on immunization dose selection of DNA vaccine B21 for therapeutic tuberculosis
10.13200/j.cnki.cjb.004519
- VernacularTitle:治疗性肺结核DNA疫苗B21免疫剂量的筛选
- Author:
DONG Jiaxin
- Publication Type:Journal Article
- Keywords:
Tuberculosis(TB);
DNA vaccine;
Immunization close;
Cellular immunity;
Humoral immunity;
Immune memory
- From:
Chinese Journal of Biologicals
2025;38(07):802-807+814
- CountryChina
- Language:Chinese
-
Abstract:
Objective To evaluate the immune responses of mice immunized with different doses of therapeutic tuberculosis DNA vaccine B21, in order to provide a reference for dose selection of quality control methods for the vaccine efficacy.Methods SPF female C57BL/6 mice were immunized by intramuscular injection of 25(LD), 50(MD), 100(HD) μg/100 μL B21 DNA vaccine through the medial thigh, combined with electroporation stimulation for a total of three times at an interval of two weeks. Three weeks after the last immunization, the spleen lymphocytes and peripheral blood serum were isolated. The cellular immunity and immune memory effects were detected by enzyme-linked immunosorbent spot(ELISpot) and flow cytometry(FCM), and the humoral immunity was detected by ELISA.Results Three weeks after the last immunization, the numbers of Ag85b-specific and Rv1738-specific IFNγ spot-forming cells(SFCs) in HD group were significantly higher than those in MD and LD groups(F = 11. 22 and 15. 89, respectively,each P < 0. 001); Rv2029c-specific IFNγ SFCs were significantly higher than that of MD group(F = 5. 761, P < 0. 05), but showed no significant difference with LD group(F = 5. 761,P > 0. 05). IL-2 SFCs of the three antigens in HD group were significantly higher than those in MD and LD groups(F =8. 044-33. 07, each P < 0. 05). The proportion of CD4~+ T cells secreting IFNγ in HD group was significantly higher than those in MD and LD groups(F = 5. 976, P < 0. 01), while there was no significant difference in the proportion of CD4~+ T cells secreting IL-2 among the three groups(F = 2. 836, P > 0. 05). The proportion of IFNγ~+ memory T cells(CD44~+ CD62L~+/~-) in HD group was significantly higher than those in MD and LD groups(F = 8. 793, P < 0. 05), and the proportion of activated memory B cells(CD19~+ CD27~+) in HD group was significantly higher than those in MD and LD groups(F = 7. 958, P < 0. 01).There was no significant difference in B21-specific IgG antibody level among the three groups(F = 5. 765, P > 0. 05).Conclusion B21DNA vaccine effectively induced cellular immunity, humoral immunity and immune memory responses in a dose-dependent manner. The dose of 100 μg/100 μL induced the best immunization effect, and ELISpot could detect IFNγ/IL-2 SFCs with good stability and reproducibility, which can be used as the recommended immune dose,evaluation indicator and detection method for quality control of B21 vaccine efficacy.
