Construction of Eukaryotic Expression Vectors Encoding CFP-10 and ESAT-6 Genes and Their Potential in Lymphocyte Proliferation

Abstract

Background: Mycobacterium (M.) bovis is the agent of bovine tuberculosis (TB) in a range of animal species, including humans. Recent advances in immunology and the molecular biology of Mycobacterium have allowed identification of a large number of antigens with the potential for the development of a new TB vaccine. The ESAT-6 and CFP-10 proteins of M. bovis are important structural and functional proteins known to be important immunogens. Methods: In the current study, the DNAs encoding these genes were utilized in the construction of pcDNA 3.1+/ESAT-6 and pcDNA3.1+/CFP-10 plasmids. After intramuscular injection of BALB/c mice with these plasmids, ESAT-6 and CFP-10 mRNA expression was assessed by RT-PCR. Mice were inoculated and boosted with the plasmids to evaluate their effects on lymphocyte proliferation. Results: Our results indicate the plasmids are expressed at the RNA level and can induce lymphocyte proliferation. Conclusion: Further study is needed to characterize the effect of these antigens on the immune system and determine whether they are effective vaccine candidates against M. bovis.

Keywords

• BALB/c mice
• CFP-10
• DNA vaccine
• ESAT-6
• Mycobacterium bovis
• PPD
• Proliferation assay