Tuberculosis and Respiratory Diseases

The Korean Academy of Tuberculosis and Respiratory Diseases (대한결핵및호흡기학회)
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Construction of Recombinant BCGs Overexpressing Antigen 85 Complex and Their Protective Efficacy against Mycobacterium tuberculosis Infection in a Mouse Model

항원 85 복합체를 과발현하는 재조합 BCG의 개발 및 마우스 모델에 있어서의 결핵균 감염에 대한 방어 효능

  • Lee, Seung-Heon (Department of Molecular Biology, Korean Institute of Tuberculosis) ;
  • Jeon, Bo-Young (Department of Microbiology, Yonsei University College of Medicine) ;
  • Park, Young-Gil (Department of Molecular Biology, Korean Institute of Tuberculosis) ;
  • Lee, Hye-Young (Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University) ;
  • Cho, Sang-Nae (Department of Microbiology, Yonsei University College of Medicine) ;
  • Kim, Hyo-Joon (Department of Biochemistry and Molecular Biology, College of Science Technology, Hanyang University) ;
  • Bai, Gill-Han (Department of Molecular Biology, Korean Institute of Tuberculosis)
  • 이승헌 (대한결핵협회 결핵연구원) ;
  • 전보영 (연세대학교 의과대학 미생물학교실) ;
  • 박영길 (대한결핵협회 결핵연구원) ;
  • 이혜영 (연세대학교 보건과학대학 임상병리과) ;
  • 조상래 (연세대학교 의과대학 미생물학교실) ;
  • 김효준 (한양대학교 과학기술대학 생화학과) ;
  • 배길한 (대한결핵협회 결핵연구원)
  • Received : 2004.03.26
  • Accepted : 2004.05.14
  • Published : 2004.08.30
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Abstract

Tuberculosis (TB) remains an enormous global health problem, and a new vaccine against TB more potent than the current inadequate BCG vaccine is urgently needed. We constructed three recombinant Mycobacterium bovis BCG (rBCG) strains over-expressing antigen (Ag) 85A, Ag85B, or both of M. tuberculosis using their own promoter and secretory sequence, or hsp60 promoter. SDS-PAGE analysis of rBCG proteins showed overexpression of Ag85A and Ag85B proteins in higher level than of those in their parental strain of BCG. In addition, rBCG(rBCG/B.FA) over-expressing Ag85A and Ag85B induced strong IFN-${\gamma}$ production in splenocytes. However, there was no significant difference in protective efficacy between rBCG and their parental BCG strain. In this study, therefore, rBCG over-expressing Ag85A, Ag85B, or both failed to show enhanced protection against M. tuberculosis infection in a mouse model.

결핵균 감염에 대한 주요 방어 항원 물질로서 항원 85 복합체(Ag85A, B, C)가 주목되고 있다. 우리는 이들 항원들을 과발현하는 재조합 BCG를 클로닝하였고, 마우스 모델을 이용하여 결핵균 감염에 대한 재조합 BCG의 방어 효능을 알아보고자 하였다. 항원 85A를 과발현하는 재조합 BCG를 rBCG/FA, 항원 85B를 과발현하는 재조합 BCG를 rBCG/FB, 그리고, 이들 두 항원을 과발현하는 재조합 BCG를 rBCG/B.FA라고 명명하였고, 이들 항원들의 과발현 여부를 SDSPAGE상에서 확인한 결과, 재조합 BCG에서 항원 85A와 B 단백질이 BCG에 비해 과발현된 것을 알 수 있었다. 면역주사한 마우스에서 분리한 비장세포를 M. tuberculosis H37Rv의 culture filtrate protein(CFP)으로 자극하여 분비된 IFN-${\gamma}$ 농도를 측정한 결과에서는 rBCG/B.FA만이 BCG에 비해 높은 IFN-${\gamma}$ 농도를 나타내었으나, 마우스 모델을 이용한 결핵균 감염에 대한 재조합 BCG의 방어 효능 실험에서는 BCG와 뚜렷한 차이를 나타내지 못하였다.

Keywords

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