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Mycobacterium tuberculosis–Secreted Protein, ESAT-6, Inhibits Lipopolysaccharide-Induced MMP-9 Expression and Inflammation Through NF-κB and MAPK Signaling in RAW 264.7 Macrophage Cells

  • Sun-Hyung Ha
  • Hyunju Choi
  • Jun-Young Park
  • Fukushi Abekura
  • Young-Choon Lee
  • Jeong-Ran KimEmail author
  • Cheorl-Ho KimEmail author
Original Article


-20pt?>Mycobacterium tuberculosis (Mtb) is a pathogenic bacterium that causes contagious tuberculosis (TB). Recently, Mtb-secreted proteins have been considered virulence factors and candidates for drugs and vaccines. Among these proteins, 6-kDa early secreted antigenic target (ESAT-6) is known to be able to induce component of matrix metalloproteinase-9 (MMP-9) in epithelial cells, leading to recruitment of macrophages. However, detailed function of ESAT-6 during macrophage recruitment to inflammatory sites remains unknown. Thus, the objective of the present study was to elucidate such function of EAST-6 and mechanism(s) involved. In the present study, we have found that recombinant ESAT-6 purified in the form of ESAT-6 double-connected structure (2E6D) could inhibit lipopolysaccharide (LPS)-induced potential of cell migration and inflammation in murine macrophage cells. Interestingly, 2E6D suppressed LPS-induced MMP-9 expression at both protein and mRNA levels as well as its enzyme activity. Levels of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) enzymes as known upregulators of MMP-9 were significantly decreased when 2E6D has been treated. In addition, nitric oxide (NO) as a second messenger was also significantly decreased by treatment with the purified 2E6D. Furthermore, 2E6D inhibited LPS-induced phosphorylation of IκB and translocation of NF-κB. Moreover, 2E6D suppressed phosphorylation of MAPK signaling proteins. Taken together, these results suggest that ESAT-6 can suppress LPS-induced MMP-9 and inflammation by downregulating COX-2, iNOS, and NO through NF-κB and MAPK signaling.

Key Words

Mycobacterium tuberculosis ESAT-6 inflammation RAW 264.7 macrophage cells 



This study has collaboratively been carried out by a member of SKKU glycobiology laboratory due to its non-financial support.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no competing interests.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Sun-Hyung Ha
    • 1
  • Hyunju Choi
    • 1
  • Jun-Young Park
    • 1
  • Fukushi Abekura
    • 1
  • Young-Choon Lee
    • 2
  • Jeong-Ran Kim
    • 3
    Email author
  • Cheorl-Ho Kim
    • 1
    Email author
  1. 1.Molecular and Cellular Glycobiology Unit, Department of Biological SciencesSungkyunkwan UniversityJangan-guRepublic of Korea
  2. 2.Faculty of Medicinal BiotechnologyDong-A UniversityBusanRepublic of Korea
  3. 3.Department of Research and DevelopmentThe Korean Institute of TuberculosisCheongju-siRepublic of Korea

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