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研究生:黃瓊慧
研究生(外文):Huang, Chiung-Hui
論文名稱:以家塵璊過敏原Derp8建立小鼠異位性皮膚炎之動物模式
論文名稱(外文):Establishment of murine model for atopic dermatitis using group 8 allergen from Dermatophagoides pteronyssinus
指導教授:蔡考圓許世明許世明引用關係許秉寧許秉寧引用關係
指導教授(外文):Chua Kaw-YanHsu Su-MingHsu Ping-Ning
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:免疫學研究所
學門:醫藥衛生學門
學類:醫學學類
論文種類:學術論文
論文出版年:1997
畢業學年度:85
語文別:中文
論文頁數:79
中文關鍵詞:異位性皮膚炎過敏原
外文關鍵詞:atopic dermatitisallergen
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早期在本實驗室中我們發現利用由日本血吸蟲所得的Glutathione-S-
tramsferase(GST)蛋白-Sj26,可在Balb/c小鼠中誘導類似人類異位性皮
膚炎(atopic dermatitis,AD)的動物模式。最近一個新的家塵璊過敏原
Der p8已經被選殖出,並且知道它與GST有相似性。因而促使我們想利用
此過敏原再次於Balb/c小鼠中呈現此動物模式。在本實驗中我們首先由
Dermatophagoides pteronyssinus cDNA library中利用PCR將Derp 8基因
選殖出,並成功的將這段基因送入Pichia yeast表現系統中,進而表現出
重組蛋白(recombinant protein)。我們利用西方墨點法(Wertern blot)
及N端牲呔序列分析證實此重組蛋白為Der p8。並利用Q管柱將Der p8純化
。利用此由yeast表現的Derp 8,以腹腔注射來免疫Balb/c小鼠,我們發
現可在Balb/c小鼠體內引發對Der p8抗原特異性IgE及total IgE上升。而
在第37天,可在小鼠的皮膚切片中觀察到有明顯的發炎細胞-噬酸性白血
球(eosinophil)及單核球 (mononuclear cell) -浸潤的情形。此皮膚的
病理變化與抗原特異性IgE或total IgE的濃度似乎沒有直接的關係。而結
果亦顯示,過敏原Der p8在引發此動物模式上扮演了一個特殊的角色。利
用此動物模式,我們可以進一步研究過敏原在AD中所扮演的角色,並探討
引發此疾病的真正機制。

In preliminary studies, we demonstrated that an inbred strain of
mice, Balb/c, when injected with recombinant Sj26 protein, a
glutathione-S-transferase of Schistosoma japonicum, developed an
inflammatory disease in skin which resembled human atopic
dermatitis. Recently, a new human allergen, Der p8, has been
cloned from Dermatophagoides pteronyssinus, and is homologous to
glutathione-S-transferase. It has been suggested that Der p8
might share similar immunogenicity as that of parasite GST, and
therefore may be able to induce atopic dermatitis change similar
to that of Sj26. In my experiments, we cloned the cDNA of Der p8
from the cDNA library of Dermatophagoides pteronyssinus using
polymerase chain reaction(PCR) and expressed the recombinant
protein in the Pichia expression system. Using Western blot and
N-terminal sequencing, we demonstrated the recombinant protein
was identical to Der p8, and we further purified this protein by
Q column. When Balb/c mices were injected with this recombinant
Der p8 intraperitoneally, Der p8 specific IgE could be detected
in the serum. At the 37th day, the skin pathology showed
inflammatory changes with infiltration of eosinophils and
mononuclear cells in the dermis layer. The development of
pathology was independent of the concentration of Der p 8
specific nor the total IgE. Our results indicated that the
allergen, Der 8, played an important role in the eliciting of
this inflammatory responses. This animal model provides a system
for further investigation of the role of allergen and the
mechanism which elicit the atopic dermatitis

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