Yonsei Med J.  2011 Nov;52(6):1008-1015. 10.3349/ymj.2011.52.6.1008.

Co-Immunization of Plasmid DNA Encoding IL-12 and IL-18 with Bacillus Calmette-Guerin Vaccine against Progressive Tuberculosis

• Jeon BY
• Eoh H
• Ha SJ
• Bang H
• Kim SC
• Sung YC
• Cho SN

• KMID: 1058823
• DOI: http://doi.org/10.3349/ymj.2011.52.6.1008

Abstract

PURPOSE
Bacillus Calmette-Guerin (BCG) vaccine has widely been used to immunize against tuberculosis, but its protective efficacy is variable in adult pulmonary tuberculosis, while it is not efficiently protective against progressive infection of virulent Mycobacterium tuberculosis strains. In this study, the protective effects of plasmid DNA vaccine constructs encoding IL-12 or IL-18 with the BCG vaccine were evaluated against progressive infection of M. tuberculosis, using mouse aerosol challenge model.
MATERIALS AND METHODS
Plasmid DNA vaccine constructs encoding IL-12 or IL-18 were constructed and mice were immunized with the BCG vaccine or with IL-12 DNA or IL-18 DNA vaccine constructs together with the BCG vaccine.
RESULTS
The BCG vaccine induced high level of interferon gamma (IFN-gamma) but co-immunization of IL-12 or IL-18 DNA vaccine constructs with the BCG vaccine induced significantly higher level of IFN-gamma than a single BCG vaccine. The BCG vaccine was highly protective at early stage of M. tuberculosis infection, but its protective efficacy was reduced at later stage of infection. The co-immunization of IL-12 DNA vaccine constructs with the BCG vaccine was slightly more protective at early stage of infection and was significantly more protective at later stage infection than a single BCG vaccine.
CONCLUSION
Co-immunization of IL-12 DNA vaccine with the BCG vaccine induced more protective immunity and was more effective for protection against progressive infection of M. tuberculosis.

Keyword

Tuberculosis; BCG; IL-12; IL-18; DNA vaccine

MeSH Terms

Animals
BCG Vaccine/*immunology
Female
Immunoenzyme Techniques
Interferon-gamma/blood
Interleukin-12/*genetics
Interleukin-18/*genetics
Mice
Mice, Inbred C57BL
Plasmids/genetics
Tuberculosis/blood/*immunology/prevention & control
Vaccines, DNA/*genetics/*immunology

Figure

• 

  Fig. 1 Construction of plasmid DNA encoding IL-12 and IL-18. Plasmid DNA vector, pGX10, is composed of a CMV promotor, TPL, SV40 late polyA tail, and an SV40 enhancer. pGX10-IL-12 was generated by insertion of murine IL-12p35, the internal ribosomal entry site (IRES) of encephalomyocarditis virus, and murine IL-12p40. pGX10-IL-18 was generated by insertion of murine IL-18. SV40, simian virus 40; pCMV, CMV promoter; TPL, tripartite leader sequences; CMV, cytomegalovirus.

• 

  Fig. 2 IFN-γ production from splenocytes by experimental vaccines. Spleno-cytes were prepared from three mice per group 30 days after the last vaccination and then stimulated with CFP antigens in vitro. The level of IFN-γ after 6 days of incubation was analyzed. The experiments were repeated three times with similar results, and the data from one representative experiment are shown. The results are expressed as the mean amounts of IFN-γ (pg/mL) of three mice (±SD) per group. p-value less than 0.05 was considered to be significant and was represented as follows: IFN-γ, interferon gamma; CFP, culture filtrate protein antigens; BCG, Bacille-Calmette-Guerin. *p<0.05, **p<0.001.

• 

  Fig. 3 Protective efficacy of experimental vaccines. Ten mice per group were immunized with the BCG vaccine singly, and co-immunized with the BCG vaccine and IL-12 DNA or IL-18 DNA vaccine. Immunized mice were infected by aerosol exposure with virulent M. tuberculosis H37Rv. The number of CFUs was measured in the lungs (A) and spleens (B) of five mice per group at 30 and 90 days post-challenge. The level of protection of experimental vaccines was calculated as CFUs in the naïve minus CFUs in the vaccinated mice. Data are represented as log10 CFU±SD, where n=5 and data is representative of two experiments. The ANOVA test was used to determine the significance of differences between experimental groups. p-value less than 0.05 was considered as significant and represented as follows; *p<0.05, **p<0.01, and ***p<0.001. BCG, bacillus Calmette-Guérin; CFU, colony forming unit; ANOVA, analysis of variance.

• 

  Fig. 4 Histopathology of lungs after aerosol infection with virulent M. tuberculosis. The right middle lobe of the lungs of mice was removed after 90 days post-challenge and lung sections were stained with H&E (×20). The representative histopathology of lungs of naïve mice (A), the BCG vaccine-immunized mice (B), co-immunized mice of the BCG vaccine and IL-12 DNA vaccine (C), and IL-18 DNA vaccine (D).

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