땅콩 알레르기의 생쥐모델에서 Bifidobacterium (strain BGN4)의 항알레르기 효과
Evaluation of Anti-allergic Effect of Bifidobacteria in Murine Model of Peanut Allergy
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목 적 : 땅콩알레르기의 생쥐모델을 이용하여 bifidobacreria(BGN4)의 위장관 투여에 의한 항원 특이적 IgE 항체 생산 조절과 비장세포의 항원 특이적 사이토카인 생성에 미치는 영향을 알아보기 위하여 본 연구를 시행하였다. 방 법 : 연구자들이 이미 확립한 IgE-매개성 땅콩 알레르기의 생쥐모델을 이용하였다. 이전의 연구 결과 가장 적절한 IgE, 생성 반응과 항원 특이적 사이토카인 생성이 유도된 땅콩 감작 농도인 1 mg/dose/mouse의 용량으로 실험 제 1, 2, 3, 7, 21일에 땅콩의 조항원을 위장관을 통하여 감작시켰다. 이 모델을 이용하여, BGN4가 제1형 알레르기 면역반응에 미치는 영향을 보기 위하여 BGN4의 경구 치료시기에 따라 실험군을 4군으로 나누어 진행하였다. 제1군은 땅콩 감작 전 2주간 BGN4를 투여한 군(pre/PN), 제2군은 땅콩 감작 전 2주 및 감작 시작 후 2주간의 총 4주간 BGN4를 투여한 군(Pre＋Post/PN), 제3군은 땅콩 감작 전 2주간 PBS만 투여한 양성대조군(PBS/PN), 제4군은 땅콩 감작 및 BGN4 치료를 하지 않은 음성 대조군(naive)이었다. 연령 4주의 female C3H/HeJ strain의 생쥐를 각 군당 3-4마리씩 이용하여 동일한 실험을 2회씩 반복하였다. 각 군별로 4군간 실험 주별 혈청 땅콩 특이 IgE 농도을 비교하였고, 실험 4주에 얻은 비장세포의 72시간 배양을 통한 땅콩 특이적 IL-4, IL-10, IL-12, IFN-γ의 생성량을 비교 분석하였다.결 과 : 땅콩의 위장관 감작이 끝난 후 땅콩 특이 IgE의 농도는 제1군(pre/PN)과 제 3군(PBS/PN)에 비하여 제2군(pre＋post/PN)에서 의미 있게 감소하였다. 그러나 땅콩 항원 자극에 따른 비장세포의 증식능은 감소하지 않았다. 한편 비장세포배양 부유액에서의 IL-4, INF-γ 및 IL-10은 제1군과 2군에서 모두 감소하였고, 특히 IL-4 의 생성 감소 및 IFN-γ/IL-4 ratio 증가는 혈청 땅콩 특이 IgE 생성의 감소와 비례하였다. IL-12는 감작 항원 자극에 의해서는 BGN4 감작 전 투여 및 감작기간 중 지속투여군인 제2군에서만 의미 있는 농도로 생성되었으며, Con A 자극에 의한 IL-12의 생성은 제2군 뿐 아니라 BGN4 전처치군인 제1군에서도 증가하였다.
Abstract
Purpose : We underwent this study to evaluate the immunomodulating effects of intragastric administration of bifidobacterium(BGN4) using murine model of peanut allergy. Methods : C3H/HeJ mice were sensitized with 1 mg/dose of crude peanut(PN) extract with cholera toxin, intragastricly. Group I mice were pretreated with BGN4 for 14 days before PN snesitization, Group II were treated 14-days each before and during sensitization, Group III were sham treated PN sensitized mice, and Group IV were naive. PN-specific serum IgE levels and PN stimulated cytokine productions from splenocyte were measured in study groups. Results : PN-specific IgE levels were significantly lowered in Group II mice compare to Group I or Group III. PN-stimulated IL-4 productions were also remarkably depressed in Group II mice. The ratio of IFN-γ/IL-4 in Group II was the highest among experimental Groups, furthermore, PN-stimulated IL-12 production was only measured in Group II mice. The decreased levels of PN-specific IgE in Group II consist with decreased production of IL-4 and increased ratio of IFN-γ/IL-4 in this experiment. Conclusion : BGN4 treatment, especially pre-and-during PN sensitization, seemed to have anti-allergic effect by suppressing PN-specific IgE production. And lowered production of IL-4, increased production of IL-12, and the increased ratio of IFN-γ/IL-4 could be suggested as a part of immunomodulating mechanism of BGN4 treatment in this experiment.
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