中文摘要
狗牙根禾花花粉為台灣及世界各地主要的禾花花粉過敏原, 而Cyn d 1是分子量34kDa的主要過敏原成份, 也是本實驗室所研究的重點。本研究室在過去已利用分子選殖的方法得知Cyn d 1可分酸性和鹼性過敏原基因。此外, 利用兩類Cyn d 1重組蛋白在動物實驗也證實兩類Cyn d 1免疫性質確有差異。
本研究進一步分析台灣地區之狗牙根花粉中Cyn d 1屬酸性或鹼性, 並以酵母菌表現的兩類Cyn d 1重組蛋白探討兩類Cyn d 1過敏原性 ( allergenicity ) 之不同。
首先分析台灣地區狗牙根花粉究竟含酸性Cyn d 1或鹼性 Cyn d 1, 以RT-PCR分析在屏東地區所採集到的花粉, 發現只具酸性Cyn d 1之mRNA, 不具鹼性之mRNA, 再用分子選殖的方法得到九株Cyn d 1 cDNA clone, 和國外酸性Cyn d 1序列做比較之後, 發現依據第157 ( S/N ) 、第181 ( P/S )及第192 ( L/Q ) 胺基酸的不同可將酸性Cyn d 1再分成兩群, 台灣地區屬於前者酸性Cyn d 1。
進而評估Cyn d 1重組蛋白的生物性功能 ( biological function ), 首先進行組織胺釋放試驗, 取四個對狗牙根花粉過敏者週邊血進行研究, 發現對於其中三人, 酸性重組蛋白在引發嗜鹼性球釋放出組織胺的能力皆較鹼性重組蛋白強。 且再用免疫競爭抑制法分析哪一種重組蛋白具較強的IgE競爭力, 發現六個病人裡, 有三個待測血清中, 酸性重組蛋白和IgE結合的能力強過鹼性重組蛋白, 其中以酸性同質異構物11號對十五位過敏病人的pool serum有最強的IgE競爭抑制力, 而鹼性同質異構物9-4-3的競爭抑制力則最低。最後用重組蛋白引起試管內淋巴細胞活化反應, 從六位病人中則看到酸鹼重組蛋白皆可刺激淋巴球增生, 刺激能力並無明顯差異。

英文摘要
Bermuda grass ( Cynodon dactylon ) pollen is a major grass pollen allergen found not only in Taiwan but also all over the world. The purpose of this study is to investigate the 34 kilodalton ( kDa ) major allergen of BGP, Cyn d 1. In the past, Cyn d 1 had been separated into the acidic and basic proteins by molecular cloning, and the immunological properties of the two groups of Cyn d 1 were different in the animal model experiment using the two groups of Cyn d 1 recombinant antigens .
In the present study, we further analyzed to which group Taiwan’s Cyn d 1 of BGP belonged. We used the two groups of recombinant Cyn d 1 proteins which were expressed by Yeast Pichia to study the difference of two groups of Cyn d 1.
First, we analyzed whether Taiwan’s BGP contains acidic Cyn d 1 or basic Cyn d 1. We used Reverse Ttranscription - Polymerase Chain Reaction to analyze Ping Tung’s BGP that we took, only mRNA of acidic Cyn d 1 was found but mRNA of basic Cyn d 1 was not. We got nine cDNA clones of Cyn d 1 by molecular cloning. According to the 157th ( S/N )、181th ( P/S ) and 192th ( L/Q ) amino acid substitution we divided acidic Cyn d 1 into two groups after being compared with foreign Cyn d 1, and Taiwan’s Cyn d 1 was the former.
Then we further evaluated the biological function of recombinant Cyn d 1 proteins. First we practiced histamine release assay to investigate peripheral blood of four patients who are sensitive to BGP. The acidic recombinant proteins were found to have better ability in inducing basophiles to release histamine than basic recombinant proteins toward three of the four patient’s peripheral blood. We further analyzed which recombinant protein had higher competitive ability by dot inhibition, and in three of the six patient’s serum, we found the IgE-binding capacity of acidic recombinant proteins were higher than basic recombinant proteins. Among these recombinant proteins toward pool serum, acidic isoform 11 was found to has the highest IgE competitive ability , and the IgE competitive ability of the basic isoform 9-4-3 was the lowest. Finally, we used recombinant proteins to elicit lymphocyte activation of six patients, not much different stimulation capacity between acidic and basic recombinant proteins was found.

目錄
中文摘要 -----------------------------------------------------------Ⅰ
英文摘要 -----------------------------------------------------------Ⅲ
緒論 ----------------------------------------------------------------- 1
材料與方法 -------------------------------------------------------- 11
結果 ----------------------------------------------------------------- 24
討論 ----------------------------------------------------------------- 28
參考文獻 ----------------------------------------------------------- 33
圖表 -----------------------------------------------------------------

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