Evaluation of protective efficacy conferred by a recombinant Mycobacterium bovis BCG expressing a fusion protein of Ag85A-ESAT-6

J Microbiol Immunol Infect. 2014 Feb;47(1):48-56. doi: 10.1016/j.jmii.2012.11.005. Epub 2013 Jan 26.

Abstract

Background: We previously constructed a recombinant bacille Calmette-Guérin (rBCG-AE) strain that could express a fused Ag85A-ESAT-6 protein. That study suggested that the rBCG-AE strain was able to induce a higher titer of antibody and elicit a more long-lived and stronger Th1-type cellular immune responses than the parental BCG strain, the rBCG-A strain (i.e., expressing Ag85A), or the rBCG-E strain (i.e., expressing ESAT-6).

Methods: In the current study, we further investigated the strain's protective efficacy against Mycobacterium tuberculosis H37Rv infection in BALB/c mice through evaluating organ bacterial loads, lung histopathology, lung immunohistochemistry, and net weight gain or loss by using conventional BCG, rBCG-A, and rBCG-E as the controls.

Results: From the 3rd to 9th weeks after the challenge infection, the bacterial counts were significantly lower in tissues (e.g., spleen and lung tissues) in the mice immunized with rBCG-AE than in the control group, but were higher than the counts in the BCG group. The pathological damage in the lung tissues of the rBCG-AE group gradually improved from the 6th to 9th weeks after being infected with M. tuberculosis H37Rv, but the score of pathological changes in the rBCG-AE group was obviously higher than the score in the BCG group. There was no difference in the percentage of IFN-γ and iNOS positive cells in the lung tissues of the rBCG-AE and BCG groups.

Conclusion: The results suggest that rBCG-AE can not promote protective efficacy against M. tuberculosis H37Rv infection, compared to the BCG vaccine.

Keywords: Ag85A; ESAT-6; Protective efficacy; Recombinant Mycobacterium bovis bacillus Calmette-Guérin (rBCG).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acyltransferases / genetics
  • Acyltransferases / immunology*
  • Animals
  • Antigens, Bacterial / genetics
  • Antigens, Bacterial / immunology*
  • BCG Vaccine / administration & dosage
  • BCG Vaccine / genetics
  • BCG Vaccine / immunology*
  • Bacterial Load
  • Bacterial Proteins / genetics
  • Bacterial Proteins / immunology*
  • Body Weight
  • Female
  • Histocytochemistry
  • Immunohistochemistry
  • Lung / immunology
  • Lung / microbiology
  • Lung / pathology
  • Mice
  • Mice, Inbred BALB C
  • Mycobacterium tuberculosis / immunology
  • Mycobacterium tuberculosis / isolation & purification
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / immunology
  • Tuberculosis / immunology
  • Tuberculosis / prevention & control*
  • Vaccines, Synthetic / administration & dosage
  • Vaccines, Synthetic / genetics
  • Vaccines, Synthetic / immunology

Substances

  • Antigens, Bacterial
  • BCG Vaccine
  • Bacterial Proteins
  • ESAT-6 protein, Mycobacterium tuberculosis
  • Recombinant Fusion Proteins
  • Vaccines, Synthetic
  • Acyltransferases
  • antigen 85A, Mycobacterium tuberculosis