Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Stimulatory Effect of Staphylococcal Protein A on Inflammatory Response in Human HaCaT Keratinocytes

사람의 피부상피세포에서 황색포도상구균의 독소인자인 Staphylococcal Protein A의 염증반응 촉진효과

  • Kwon, Hyun-Jin (Department of Laboratory Medicine, Daegu Fatima Hospital) ;
  • Kim, Yeon-Jung (Seoul Medical Center Research Institute, Seoul Medical Center) ;
  • Jang, Sung-Hee (Seoul Medical Center Research Institute, Seoul Medical Center) ;
  • Bae, Bo-Kyoung (Research Institute for Veterinary Science, College of Veterinary Medicine, Seoul National University) ;
  • Youn, Hwa-Young (Research Institute for Veterinary Science, College of Veterinary Medicine, Seoul National University) ;
  • Lee, Hee-Woo (Research Institute for Veterinary Science, College of Veterinary Medicine, Seoul National University)
  • 권현진 (대구파티마 병원 진단검사실) ;
  • 김연정 (서울의료원 부설연구소) ;
  • 장성희 (서울의료원 부설연구소) ;
  • 배보경 (서울대학교 수의과대학 수의과학연구소) ;
  • 윤화영 (서울대학교 수의과대학 수의과학연구소) ;
  • 이희우 (서울대학교 수의과대학 수의과학연구소)
  • Received : 2011.12.07
  • Accepted : 2011.12.16
  • Published : 2011.12.31
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Abstract

Staphylococcus aureus is a major human pathogen that is associated with various types of local and systemic infection. Staphylococcal protin A (SPA), a highly expressed surface component of S. aureus, may have a role in virulence such as activating inflammation and interfering with immune clearance. We examined the effect of recombinant SPA on inflammatory response in human HaCaT keratinocytes. The recombinant SPA protein was prepared using the pET-28a Vector System in Escherichia coli. The expression of pro-inflammatory related adhesion molecules and cytokines in HaCaT cells incubated for 6, 12, and 24 h with SPA (2 ${\mu}g$/ml) was analyzed by comparative RT-PCR or ELISA. The expression of E-selectin, ICAM-1, MCP-1, IL-6 and IL-8 was significantly increased in HaCaT from 6 to 24 h after treatment with SPA. SPA showed the effect on the adhesion-promoting ability of U937 monocytes to HaCaT cells. Our data demonstrate that SPA stimulates inflammatory response of HaCaT cells, implicating an important factor for exacerbation of skin inflammation of immunologic disease.

황색포도상구균은 사람에게서 염증을 동반한 다양한 형태의 국소적 또는 전신적 감염을 일으키는 주요 병원균이며, 황색포도상구균에서 풍부하게 발현되는 Staphylococcal protein A (SPA)는 염증의 활성화나 면역 반응의 회피와 관련된 균력인자로서 작용할 수 있다. 본 연구에서는 사람의 HaCaT 피부상피세포에서 재조합 SPA 단백질을 이용하여 염증반응에 대한 효과를 조사하기 위해서 pET-28a 발현벡터시스템을 이용하여 성공적으로 재조합 SPA 단백질을 제작하였고, 이 단백질(2 ${\mu}g$/ml)을 6, 12 및 24시간 처리한 HaCaT 피부상피세포에서 RT-PCR 및 ELISA를 이용하여 염증관련 부착인자 및 사이토카인의 발현을 분석하였다. SPA 처리 후 6시간에서 24시간까지 E-selectin, ICAM-1, MCP-1, IL-6 및 IL-8의 발현이 현저하게 증가함을 확인하였다. 또한 SPA는 HaCaT 피부상피세포에 대한 U937 단핵구의 부착력을 증진시켰다. 따라서, 본 연구의 결과는 SPA가 HaCaT 피부상피세포의 염증반응을 촉진시킨다는 사실을 보여주었으며, 황색포도상구균에 의한 피부염증질환에 있어서 중요한 병원성인자로서의 역할을 수행한다는 사실을 시사해준다.

Keywords

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