Oral Immunogenicity of Plant-MadeMycobacterium tuberculosisESAT6 and CFP10

Elena A. Uvarova,Olesya V. Nosareva,Alla A. Zagorskaya,Natalya V. Permyakova,Almagul A. Kakimzhanova,Pavel A. Belavin,Elena V. Deineko

doi:10.1155/2013/316304

Abstract

Two lines of transgenic carrot plants producing Mycobacterium tuberculosis proteins (ESAT6 and CFP10) have been constructed. The target proteins are present in carrot storage roots at a level not less than 0.056% of the total storage protein (TSP) for ESAT6 and 0.002% of TSP for CFP10. As has been shown, oral immunization of mice induces both the cell-mediated and humoral immunities. These data suggest that the proteins in question are appropriate as a candidate edible vaccine against tuberculosis.

Highlights

The tuberculosis caused by the bacterium Mycobacterium tuberculosis is currently a topical problem in many countries of the world
The following primers were used for amplifying the genes esat6 and cfp10 using the M. tuberculosis genomic DNA as a template: esatupper 5󸀠-GCTCTAGAATGACAGAGCAGCAGTGGAATTTCGCGG-3󸀠 and esat-low 5󸀠-CGGGATCCCTATGCGAACATCCC-3󸀠 for the esat6 gene and for the cfp10 gene, cfpupper 5󸀠-GCTCTAGAATGGCAGAGATGAAGACC-3󸀠, and cfp-low 5󸀠-CGGGATCCGAATTCTCAGAAGCC-3󸀠
The fragments of esat6 (288 bp) and cfp10 (303 bp) genes were hydrolyzed with the restriction endonucleases XbaI and BamHI, purified using a GenElute HP Plasmid Midiprep kit (Sigma-Aldrich, United States), and ligated with the pBI121 preliminary hydrolyzed at the same sites

Summary

Introduction

The tuberculosis caused by the bacterium Mycobacterium tuberculosis is currently a topical problem in many countries of the world. The number of tuberculosis cases, including fatal outcomes, has been ever increasing [1]. The recorded dynamics of tuberculosis morbidity are complicated by the spread HIV and hepatitis, ascribed by the WHO to socially significant diseases [2]. The Bacillus Calmette-Guerin (BCG) vaccine—a live attenuated M. bovis strain [3]—was designed as early as the 1920s; by the end of the 20th century, about 3 billion persons worldwide had been vaccinated [4, 5]. Note that the genome of all the M. bovis BCG strains lacks the RD1 region, characteristic of M. tuberculosis; this region houses very important virulence factors, such as ESAT6 and CFP10 [6, 7]. Despite the facts that BCG as a prevention vaccine is used for protection of uninfected children and is inefficient for adults for the most abundant tuberculosis form, lung tuberculosis, BCG still remains the only available vaccine against tuberculosis [3]

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