가와사끼병에서 가용성 ELAM-1에 관한 연구

Abstract

[한글]

Kawasaki병은 혈관염의 임상양상을 나타내는 급성 전신성 질환으로 현재까지 그 원인에 대한 연구가 많이 진행되었으나 확실한 병인론은 밝혀진 바가 없다. 근자에 들어서 혈관내피세포가 활성화 될 때 세포표면에 발현되는 여러가지 항원들 중에서 특히 혈중의 백혈

구와 결합하는 intercellular adhesion molecules에 관한 연구가 활발히 진행되면서 혈관내피세포에서만 발현되는 endothelial leukocyte adhesion molecule-1 (ELAM-1)에 대한 관심이 증가하고 있다. 더우기 ELAM-1은 가용성 상태로 세포표면에서 유리되어 혈중에서 발견될 수 있어서, 가용성 ELAM-1의 측정은 염증성 질환에서 혈관염의 정도를 측정하는데 유용한 것으로 보고되고 있다. 저자는 급성 전신성 혈관염인 Kawasaki병에서 가용성 ELAM-1치를 측정하고 아울러 C-reactive protein(CRP), tumor necrosis factor-α(TNF-o), interleukin-6(IL-6) 등을 측정하여 급성기에 보이는 혈관염과의 연관성을 살펴보고, 아급성기로 이행함에 따른 변화를 조사하였다.

1992년 9월부터 1993년 6월까지 세브란스병원 소아과에 입원하였던 Kawasaki병 환아 24명, 동기간에 입원한 Henoch-Schonlein자반병 환아 10명, Behcet병 환자 18명, 정상소아 10명을 대상을 하였고, Kawasaki병에서는 급성기 및 아급성기 혈청을 얻어 -70"C에 보관

하였다가 실험 직전에 실온에 녹여 사용하였다.

가용성 ELAM-1은 Bender MedSystems의 ELISA kit를 이용하여 측정하였고, TNF-α는 R&D Systems사의 Quantikine***TM ELISA kit를 사용하여 측정하였으며, IL-6는 Fujirebio Inc.사의 ELISA kit를 이용하여 측정하였다.CRP는 fluorescence polarization immunoassay

방법으로 측정하였다.

그 결과 혈청 가용성 ELAM-1은 급성기에 27.5 ng/ml로 현저한 증가를 보이다가 아급성기에는 16.2 ng/ml로 감소하였다. TNF-α는 급성기에 44.1pg/ml로 증가하였다가 아급성기에는 24.0 pg/ml로 현저하게 감소하였다. 대조군으로 Henoch- Schonlein자반병에서 혈청

가용성 ELAM-1은 12.9 ng/ml로 정상(6 ng/ml이하)과 비교하여 증가되어 있었으나 급성기 Kawasaki병에서처럼 증가하지는 않았다. 또한 Behcet병에서는 5.5 ng/ml로 정상에 비하여 증가된 것을 관찰할 수 없었다. Kawasaki병 환아의 혈청 가용성 ELAM-1치는 혈중 TNF-α

와 직선적인 상관관계를 보이고 있었으며, 아울러 혈중 IL-6치와도 상관관계를 보였다 그러나 혈중 CRP나 ESR과는 상관관계를 보이지 않았다.

이처럼 Kawasaki병에서 가용성 ELAM-1이 혈중에 증가되는 것은 혈관의 염증상태의 정도와 비례하는 것으로 사료되며 혈관내피세포의 표면에 표현되는 ELAM-1이 Kawasaki병에서 병인론과 관계가 있는 지에 대해서는 더 많은 연구가 있어야 될 것으로 사료된다.

Serum soluble endothelial leukocyte adhesion molecule-1 level in Kawasaki disease

Wan Kyu Lee

Department of Medical Science The Graduate School, Yonsei University

(Directed by Assistcnt Professor Dong Soo Kim)

Kawasaki disease is an acute systemic disease with clinical manifestions of

vasculitis. There have been many studies concerning the etiology and pathogenesis

of the disease. But they have not yet been clarified. Rececntly among various

surface antigens expressed on the cell membrane during the activation of the

vascular endothelial cells, the intercellular adhesion molecules which bind with

the peripheral blood leukocytes have been studied a great deal. Especially the

endothelial leukocyte adhesion molecule-1(ELAM-1), which is expressed only on the

vascular endothelial cells, is gathering interests of many researchers. The ELAM-1,

which can be detected in the blood as a soluble antigen shedded from the cell

membrane, could be used as an index of severity of the vasculitis in inflammatory

diseases. In this study the serum soluble ELAM-1 levels along with C-reactive

protein(CRP), tumor necrosis factor-α(TNF-α), and interleukin-6(IL-6) levels have

been determined in the patients with Kawasaki disease to see the relationship with

the degree of vasculitis.

The study subjects were 24 patients with Kawasaki disease, 10 with

Henoch-Schonlein purpura, and 18 with Behcet's disease who were admitted to the

Department of Pediatrics, Severanc Hospital between September 1972 and June 1993.

Ten children who were admitted for elective surgery consisted of the negative

control group. The serum samples were obtained during the acute and subacute

stages, stored frozen at -7O°C, and thawed at room temperature just before the

measurement.

The serum soluble ELAM-1 levels were measured by the sandwich ELISA method(Bender

MedSystems), TNF-α levels by the sandwich ELISA method(R&D System), and IL-6

levels by the sandwich ELISA method(Fujirebio Inc.). CRP was measured by the

fluorescence polorization immunoassay method.

The serum soluble ELAM-1 level rapidly rose up to 27.5 ng/ml during the acute

stage and fell to 16.2 ng/ml during the subacute stage. Similary TNF-α level

increased to 44.1 pg/ml during the acute stage and significantly decreased to 24.0

pg/ml during the subacute stage. In the patients with Henoch-Schonlein purpura, the

serum soluble ELAM-1 level was elevated to 12.9 ng/ml, which was higher as compared

to the normal elvel(less than 6 ng/ml) but not as high as that in the patients with

Kawasaki disease. For Behcet's disease it was 5.5 ng/ml; that is , not at all

higher than the normal level. There was a linear relationship between the serum

soluble ELAM-1 levels of the Kawasaki patients and the serum TNF-a or IL-6 levels.

However, the serum CRP or ESR did not show any direct relationship with the serum

soluble ELAM-1 levels.

As the results suggest , the serum soluble ELAM-1 levels increase in proportion

with the degree of vasculitis. And whether the ELAM-1, which is expressed on the

cell surface of the vascular endothelium, is related to the pathogenesis of

Kawasaki disease must further be studied.

[영문]

Kawasaki disease is an acute systemic disease with clinical manifestions of vasculitis. There have been many studies concerning the etiology and pathogenesis of the disease. But they have not yet been clarified. Rececntly among various surface antigens expressed on the cell membrane during the activation of the vascular endothelial cells, the intercellular adhesion molecules which bind with the peripheral blood leukocytes have been studied a great deal. Especially the endothelial leukocyte adhesion molecule-1(ELAM-1), which is expressed only on the

vascular endothelial cells, is gathering interests of many researchers. The ELAM-1, which can be detected in the blood as a soluble antigen shedded from the cell membrane, could be used as an index of severity of the vasculitis in inflammatory diseases. In this study the serum soluble ELAM-1 levels along with C-reactive protein(CRP), tumor necrosis factor-α(TNF-α), and interleukin-6(IL-6) levels have been determined in the patients with Kawasaki disease to see the relationship with the degree of vasculitis.

The study subjects were 24 patients with Kawasaki disease, 10 with Henoch-Schonlein purpura, and 18 with Behcet's disease who were admitted to the Department of Pediatrics, Severanc Hospital between September 1972 and June 1993.

Ten children who were admitted for elective surgery consisted of the negative control group. The serum samples were obtained during the acute and subacute stages, stored frozen at -7O°C, and thawed at room temperature just before the measurement.

The serum soluble ELAM-1 levels were measured by the sandwich ELISA method(Bender MedSystems), TNF-α levels by the sandwich ELISA method(R&D System), and IL-6 levels by the sandwich ELISA method(Fujirebio Inc.). CRP was measured by the fluorescence polorization immunoassay method.

The serum soluble ELAM-1 level rapidly rose up to 27.5 ng/ml during the acute stage and fell to 16.2 ng/ml during the subacute stage. Similary TNF-α level increased to 44.1 pg/ml during the acute stage and significantly decreased to 24.0 pg/ml during the subacute stage. In the patients with Henoch-Schonlein purpura, the serum soluble ELAM-1 level was elevated to 12.9 ng/ml, which was higher as compared to the normal elvel(less than 6 ng/ml) but not as high as that in the patients with Kawasaki disease. For Behcet's disease it was 5.5 ng/ml; that is , not at all higher than the normal level. There was a linear relationship between the serum soluble ELAM-1 levels of the Kawasaki patients and the serum TNF-a or IL-6 levels.

However, the serum CRP or ESR did not show any direct relationship with the serum soluble ELAM-1 levels.

As the results suggest , the serum soluble ELAM-1 levels increase in proportion with the degree of vasculitis. And whether the ELAM-1, which is expressed on the cell surface of the vascular endothelium, is related to the pathogenesis of Kawasaki disease must further be studied.