H. Serum IL-6 and IL-17 levels were down-regulated after transfer
Ⅴ. CONCLUSION
We confirmed the frequency of CD4+CD25+T cells in BD-like mice was lower than BDN mice. CD4+CD25+T and Treg cell levels by primary culture of splenocytes were lower in BD-like mice compared to BDN mice. The frequencies of CD4+CD25+T and Treg cells in co-cultures from normal CD4+T cells isolated from splenocytes with normal mDC was higher than CD4+T cells from splenocytes of BDN and BD-like. In vivo, the frequency of CD4+CD25+T cells were higher in transfer with 3 x 105CD4+CD25+T cells in BD-like mice compared to transfer with 3 x 103and 3 x 104CD4+CD25+T cells. In addition, transfer with CD4+CD25+T cells to BD-like mice improved the BD-like symptoms. The severity scores were accordingly decreased compared to transfer of CD4+CD25-T cells to BD-like mice. After 3 x 105 CD4+CD25+T cells transfer, the severity score of BD-like mice was decreased compared to before the transfer. The serum protein levels of TGF-β and IL-10 in BD-like mice transferred with 3 x 105 CD4+CD25+T cells were higher, but IFN-γ, TNF-α, IL-6, and IL-17 were lower than not transferred BD-like mice. The mRNA level of TGF-β was also affected in the transfer of CD4+CD25+T cells. TGF-β was higher than not transferred BD-like mice. Whereas IFN-γ, TNF-α, IL-17A, IL-17F, and ROR-γt of transferred with CD4+CD25+T cells were lower than not transferred BD-like mice.
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국문요약
-단순포진 바이러스로 유도한 베체트병 마우스모델에서 CD4+CD25+ 조절 T 세포의 역할
아주대학교 대학원 의생명과학과 심 주 아
(지도교수: 손 성 향)
베체트병은 만성 염증질환으로 재발성 구강 궤양, 외음부 궤양, 피부 및 안 증상 등이 특징인 질병이다. 베체트병은 여러 증상들이 동시에 발병하거나 초기 피부 점막증상이 호전과 악화를 거듭하다 관절 증상, 안 증상, 신경계 증상까지 발전할 수 있다. 특히 안 증상은 대표적인 베체트병 주 증상으로 실명이라는 심각한 후유증을 초래할 수 있다. 이러한 심각한 증상을 변화시킬 수 있는 방법으로, 본 연구에서는 T 세포의 한 부분으로 알려져 있는 CD4+CD25+ 조절 T 세포의 역할에 대하여 알아보고자 한다. CD4+ CD25+ 조절 T 세포는 자기 와
비자기 항원에서 잘못된 면역반응을 예방할 수 있고 말초조직에서
self-tolerance 를 유지시켜 준다. 그리고 자가면역을 조절하고 병을 일으킬 수 있는 림프구를 억제하여 자가면역을 예방할 수 있다고 알려져 있다. 이와 같은
보고를 바탕으로 단순포진 바이러스로 유도한 베체트병 마우스모델에서
CD4+CD25+ 조절 T 세포의 연관성을 알아보고자 한다.
베체트병 마우스모델의 비장에서 CD4+CD25+ 조절 T 세포의 발현 양상을 알아보고 CD4+CD25+T 세포를 꼬리 정맥을 통하여 transfer 하여 베체트병 증상에 어떠한 영향을 주는지 관찰하였다. 2 주 후 transfer 한 베체트병 마우스의 비장에서 CD4+CD25+T 와 조절 T 세포의 발현을 유세포 분석기로 확인하였다.
그리고 혈청을 얻어 ELISA 를 통하여 사이토카인의 변화를 확인하였고, RT-PCR 을 통하여 사이토카인의 mRNA 수준을 비교하였다.
그 결과 베체트병 마우스모델의 증상 군이 무증상 군 (바이러스 접종 후 증상이 나타나지 않은 군) 에 비하여 CD4+CD25+ 조절 T 세포가 낮게 발현되었다. 그
발현의 차이가 증상의 변화에 관련이 있는지 알아보기 위하여 베체트병
마우스모델에 CD4+CD25+T 세포를 주입하여 비장에서 조절 T 세포의 변화를 알아본 결과 3 x 103 과 3 x 104 CD4+CD25+T 세포를 주입한 3 x 105 를 주입한
군에서 조절 T 세포가 증가하는 것을 확인하였다. 또한 3 x 105 CD4+CD25+T
세포를 주입한 군에서 증상이 호전되는 것을 관찰하였으며 severity score 역시
유의하게 감소되었다. 사이토카인의 혈청단백질 변화를 측정한 결과 주입하지
않은 베체트병 마우스모델 보다 3 x 105 CD4+CD25+T 세포를 주입한 마우스
모델에서 TGF-β 와 IL-10 이 증가하였고 IFN-γ, TNF-α, IL-6 와 IL-17 은 유의하게 감소한 결과를 보였다. mRNA 역시 단백질 수준과 비슷한 결과를 나타냈다.
이러한 결과를 바탕으로 CD4+CD25+ 조절 T 세포의 발현 저하가 베체트병
마우스모델에서 증상을 유발시키는 요인으로 작용하는 것으로 보이고,
핵심어 : CD4+CD25+T 세포, 조절 T 세포, 베체트병 마우스모델