Korean Journal of Clinical Laboratory Science (대한임상검사과학회지)

Korean Society for Clinical Laboratory Science (대한임상검사과학회)
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The Development of Diagnosis for Atopic Dermatitis by Evaluating the Expression of Skin Barrier Proteins Using a Non-Invasive Method

비침습적 피부장벽단백질 측정을 통한 아토피 피부염의 진단 및 유용성

  • Kim, In Sik (Department of Biomedical Laboratory Science, School of Medicine, Eulji University) ;
  • Lee, Ji-Sook (Department of Clinical Laboratory Science, Wonkwang Health Science University)
  • 김인식 (을지대학교 대전캠퍼스 임상병리학과) ;
  • 이지숙 (원광보건대학교 임상병리과)
  • Received : 2017.10.11
  • Accepted : 2017.10.31
  • Published : 2017.12.31
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Abstract

Atopic dermatitis is a chronic inflammatory skin disease caused by a variety of genetic and environmental factors, dysregulation of immunological response, as well as dysfunction of the skin barrier proteins. The purpose of this study is to develop an ELISA kit suitable for evaluating the expression of skin barrier proteins. Proteins were obtained from the skin via AriNo and D-Squame patches. The efficiency of protein collection from the skin, using the Arino patch, was shown to be more effective than using D-Squame; while the efficiency of lysis using 0.1% Triton-X100 was higher than that of other lysis solutions, including 0.1 M Tris-HCL, 0.1% Tween-20, and 5 mM KOH. Recombinant skin barrier proteins, such as filaggrin and involucrin, were produced by molecular biological methods. Monoclonal antibodies against filaggrin and involucrin were produced by immunization of mice, fusion of spleen cells and myeloma cells, as well as a selection of antibody-producing hybridoma cells. The filaggrin expression in the skin of subjects suffering from atopic dermatitis was lower than that in normal mice. Involucrin expression was not altered between normal individuals and subjects with atopic dermatitis. These findings contribute to an elucidation of the importance of the skin barrier protein expression in atopic dermatitis and the development of a diagnostic kit for atopic dermatitis.

아토피 피부염은 만성염증 피부질환으로서 유전적 요소, 환경적 요소, 면역반응의이상, 피부장벽단백질의 기능 이상에 의하여 발생한다. 본 연구의 목적은 피부장벽단백질의 발현을 측정할 수 있는 ELISA 키트 개발에 있다. AriNo와 D-Squame 패치를 이용하여 비침습적으로 피부에서 단백질을 얻을 수 가 있었고, AriNO가 좀 더 많은 단백질을 획득하였다. 0.1% Triton X-100용액이 다른 용해용액인 0.1 M Tris-HCL, 5 mM KOH, 0.1% Tween-20보다 높은 단백질 수율을 나타냈다. 피부장벽 단백질 측정을 위한 ELISA 키트 개발을 위하여 분자생물학적인 방법을 이용하여 필라그린과 인보루크린의 재조합단백질을 생산하였고, 이에 대한 단일클론항체를 면역학적인 방법을 이용하여 만들었다. 아토피 피부염 환자의 피부에서는 필리그린의 발현이 유의하게 줄어들었고, 인보루크린은 정상인과 아토피 피부염 환자에서 차이를 나타내지 않았다. 본 연구의 결과를 통하여 아토피 피부염에서 피부장벽단백질의 중요성을 규명하였고, 향후 아토피 피부염의 진단키트를 개발하는데, 유용할 것이다.

Keywords

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