臺灣博碩士論文加值系統

花粉為大氣環境中已知過敏原，並且與世界各國氣喘、過敏性鼻炎之盛行率及死亡率不斷攀升有關。前期監測大氣中花粉種類與濃度的結果顯示，構樹花粉為台南地區大氣中最主要的花粉組成，約佔40％，部份研究也認為一些過敏症狀的盛行率與暴露到構樹花粉有關。因此，本研究欲透過大氣構樹花粉濃度與醫院門診人次的相關性分析，以及萃取構樹蛋白質進行臨床過敏原試驗，來評估構樹花粉在台灣地區是否為一潛在的新過敏原。本研究以孢粉採樣器收集大氣中構樹花粉以進行微粒計數，在光學顯微鏡下，花粉型態呈現約10 μm-12 μm粒徑範圍，其粒徑小於一般花粉，顯示可能容易被吸入。此外，2005年構樹花粉量相較於前期研究有減少之趨勢(31.9%)。進一步地，利用鄰近監測站之大型醫學中心的每日各類疾病之住院人次與門診人次資料，以及台南空氣污染測站每日數據，進行相關性的探討；初步模式分析結果顯示，當每週空氣中構樹花粉濃度上升100 particles/m3，呼吸道疾病門診(RR=1.142)、過敏性鼻炎門診(RR=1.206)、氣喘門診(RR=1.160)及呼吸道疾病住院人次(RR=1.137)相對危險性均會顯著增加，各類相關疾病門診人次增加之相對危險性中，又以過敏性鼻炎門診增加之比例最高。延續著區域性資料分析的結果，本研究進一步地嘗試透過臨床的血清及皮膚試驗來證明構樹花粉的致敏性。在花季（3至5月）採集新鮮構樹花粉，將採集的新鮮花粉及市售花粉分別進行花粉蛋白質之萃取及定量。試驗族群選以20位過敏性學童（總免疫球蛋白E之中位數為580.50 EU/L及20位健康學齡孩童（總免疫球蛋白E之中位數為17.65 EU/L），共21位男性及19位女性，年齡為介於7-12歲之孩童血清樣本，以酵素免疫分析法 (Enzyme-Linked ImmunoSorbent Assay; ELISA)進行花粉過敏原之特異性免疫球蛋白E (Specific IgE)試驗，並以市售花粉為参考組。此外招募30位自願受試者（12位男性及18位女性，年齡為介於23-38歲之成年人），進行皮膚試驗(Skin prick test; SPT)及特異性花粉過敏原之免疫球蛋白E試驗。以1:100 (w/v) 萃取新鮮花粉，其平均可溶性蛋白質濃度為0.29 mg/ml。比較學齡孩童之試驗族群於新鮮花粉與市售花粉過敏原萃取物之特異性免疫球蛋白E試驗結果顯示具顯著相關性 (spearman correlation= 0.89, p<0.01)。過敏性之學齡孩童吸光值(optical density; OD)高於健康學童 (median value 0.035 v.s 0.025, p<0.05)，皮膚試驗結果呈現，構樹過敏原濃度為10 μg/ml進行皮膚試驗後，反應呈現陽性及陰性兩族群之總免疫球蛋白E之中位數分別為120.0 ku/L及50.6 ku/L，特異性免疫球蛋白E之中位數分別為0.064及0.027，此結果呈現出構樹花粉過敏原之致敏率約為46%。構樹花粉的致敏特性經由特異性花粉過敏原之免疫球蛋白E試驗及皮膚試驗結果顯示，構樹花粉為台灣地區重要之新過敏原。

Pollens are known as major allergens in the atmosphere while increasing prevalence and mortality of asthma and rhinitis has been observed around the world. Previous pollen monitoring in Tainan City of Taiwan had revealed that Broussonetia papyrifera could be a major genus, about 40%, in Taiwan, and literatures have reported that allergic symptoms could be induced by exposure to pollens of Broussonetia papyrifera. This study was therefore aimed to assess whether Broussonetia papyrifera pollen in atmosphere could be a seasonal allergen in Taiwan. Daily pollen counts were achieved by collecting pollen spores using a Burkard trap. The size of Broussonetia papyrifera pollens, between 10μm-12μm, was smaller than many prevalent types of pollen, and might be easier to penetrate through respiratory track after inhalation. Our study has analyzed first, using generalized addictive models (GAM) with Poisson regression model, the associations between the levels of daily Broussonetia concentration, about 32% of the total pollens measured, and frequencies of daily clinic visits and hospital admission in a major teaching hospital from 1 January to 31 December in 2005. Time-series analysis, having filtered the seasonal trends and adjusted for the day-of the week cycles, weather factors, and air pollutants, was conducted to determine the influence of ambient spores on asthma hospital admissions. The strongest association was observed for effect of ambient Broussonetia concentrations on the number of clinical visits hospital admission (respiratory tract disease clinic visits (RR=1.142), allergic rhinitis clinic visits (RR=1.206), Asthma clinic visits (RR=1.160) and respiratory tract disease hospital admission (RR=1.137). Fresh pollens of Broussonetia papyrifera were then collected from trees during March to May, the flowering season, for extraction before identical analysis was conducted with commercial pollens (Allergen AB, Sweden) for comparison. Serum of 20 allergic asthmatics (median level of total IgE=580.50 EU/L) and 20 healthy schoolchildren (median level of total IgE= 17.65 EU/L), together of 40 with 21 males and 19 female aged between 7~12 years old, were analyzed for the specific IgE levels after challenged with Broussonetia papyrifera extract, both of fresh ones and commercial preparation. Furthermore, 30 volunteer subjects (12 males and 18 female; aged between 23~38 years old) were recruited for skin prick test (SPT) and the specific IgE analysis to Broussonetia papyrifera by Enzyme-Linked ImmunoSorbent Assay (ELISA). The mean soluble protein content of the fresh pollens extract of was observed at 0.29 mg/ml of the 1:100 (w/v) concentrations. Specific IgE levels measured in subjects when exposed to fresh pollen extract correlated well with those measured after exposure to commercial pollen extract (spearman correlation= 0.89, p<0.01). OD value of specific IgE levels in schoolchildren with allergic asthma was higher than that of healthy children (median value 0.035 v.s 0.025, p<0.05). Using sera from a general population, among 3 different allergen levels used for SPT, the only significant difference, when compared the negative versus the positive response group, that could be consistently supported by the measurements of total IgE (median value of 50.6 ku/L for negative and 120.0 ku/L for positive SPT) and specific IgE level (median value of 0.027 for negative and 0.064 for positive SPT) was at the concentration of 10 μg/ml (46% sensitized rate). The allergic characteristic of Broussonetia papyrifera pollens is supported by tests on both school-aged children as well as young adults based on not only the specific IgE analysis but also the skin-prick-test, and therefore suggested to be one important emerging allergen in Taiwan.

中文摘要 I
英文摘要 I
致謝 III
目錄 I
表目錄 III
圖目錄 IV
第一章 前言 1
1-1 研究緣起 1
1-2 研究目的 2
第二章 文獻回顧 3
2-1 過敏原與過敏性疾病 3
2-2 花粉過敏原 4
2-2-1 氣候條件 6
2-2-2 空氣污染物 6
2-3 各國花粉與過敏性疾病之研究 7
2-3-1 致敏花粉之地域性 7
2-3-2 新致敏花粉 9
2-3-3 新致敏花粉之確認 9
2-4 台灣有關花粉與其致敏性之研究 10
2-4-1 花粉曆研究 10
2-4-2 台灣地區花粉之主要組成 11
2-5 構樹與過敏性症狀之相關研究 11
2-6 小結 12
第三章 研究材料與方法 13
3-1 研究架構 13
3-2 空中花粉與相關疾病門診及住院人次之探討 13
3-2-1 花粉採樣方法 14
3-2-2 花粉鑑定方式 15
3-2-3 資料收集 16
3-3 過敏原之確認 16
3-3-1 花粉收集與保存 16
3-3-2 花粉蛋白質萃取及定量 17
3-3-3 受試者選取 19
3-3-4 特異性過敏原測試 (Specific IgE) 21
3-3-5 臨床皮膚穿刺試驗 (Skin prick test) 21
3-4 統計分析方法 22
3-4-1 空中花粉與相關疾病門診及住院人次之探討 22
3-4-2 過敏原之確認 23
第四章 研究結果 24
4-1 空中花粉濃度分布對過敏性呼吸道疾病門診及住院人次之貢獻 24
4-1-1 花粉採樣及鑑數結果 24
4-1-2 空中花粉濃度與門診及住院人次之相關性 25
4-2 構樹花粉致敏原之確認 28
4-2-1 構樹花粉型態 28
4-2-2 花粉蛋白質萃取及定量 28
4-2-3 學齡孩童之特異性過敏原測試 28
4-2-4 成人受試者之試驗結果 29
第五章 研究討論 33
5-1 空中花粉濃度分布對過敏性呼吸道疾病門診人次之貢獻 33
5-1-1 花粉採樣及鑑數結果 33
5-1-2 空中花粉與相關疾病門診人次之相關性 34
5-2 構樹花粉致敏原之確認 35
5-2-1 學齡孩童之特異性過敏原測試 35
5-2-2 成人受試者之試驗結果 35
5-2-3 皮膚試驗過敏原濃度之選擇 37
5-2-4 花粉之致敏機制 38
5-3 研究限制 39
第六章 研究結論與建議 40
参考文獻 42
附錄 76
附錄一 同意臨床試驗證明書 76
附錄二 優良臨床試驗規範訓練結業證書 77
附錄三 國立成功大學醫學院附設醫院人體試驗委員會臨床試驗申請表 78
附錄四 國立成功大學醫學院附設醫院臨床試驗說明及同意書 81
自述 83

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