臺灣博碩士論文加值系統

D 型肝炎病毒 ( hepaptitis delta virus, HDV )為一缺陷型病毒，必
需與B 型肝炎病毒 ( hepaptitis B virus, HBV )同時感染 ( coinfection )
或重複感染 ( superinfection ) 方可完成生活史。合併感染HBV 及
HDV 的病患，形成慢性肝炎、猛爆性肝炎、肝硬化、甚至肝癌的比
例均較單獨感染HBV 為高，但其分子層次的機制不明。因HDV 可
產生兩種大小不同的蛋白：large delta antigen ( LDAg )和small delta
antigen ( SDAg )，懷疑此兩抗原有可能與宿主細胞的分子作用而產生
症狀。本實驗室利用秀麗隱桿線蟲( Caenorhabditis elegans, C. elegans )
生活史短、容易操作，並有清楚的遺傳學建立分子途徑的特色建立研
究病毒之平台。在先前的實驗中發現：在轉殖線蟲咽喉部 ( pharynx )
表現HDV 的SDAg 會使線蟲有生長遲緩，及生育力降低的現象。本
實驗為了知道此抗原何段具有作用，將不同長度之SDAg 分別命名為
SD441、SD327、SD267、SD201 表現在線蟲的pharynx，並比較不同
基因型的SDAg，以及在不同發育階段表現SDAg 的結果有何差異，
藉以研究此蛋白對線蟲的影響。發現 (1) 除了SD201 此不具
nuclear-localization signal ( NLS )的SDAg 外，其餘的對線蟲有顯著影
響，(2) 在線蟲L4 時於pharynx 表現SDAg 會對線蟲造成最大影響，
(3) 對線蟲造成影響的強弱為：第二基因型大於第一基因型大於第三
基因型。以上結果顯示SDAg 需進入核內干擾核內未知分子作用才能
對線蟲造成影響。未來若能找到此分子，則可回推是否在人類也有同
源基因，而有助了解HDV 引起症狀的真相。

It is known that hepatitis delta virus (HDV) requires hepatitis B
virus (HBV) to complete its life cycle. When patients are coinfected or
superinfected with HBV and HDV, it causes fulminant hepatitis and
progressive chronic liver cirrhosis, indicating HDV plays an important
role that makes the symptoms become more seriously. The mechanism by
which HDV infection contributes to clinical hepatitis is poorly
understood. HDV has two different size of proteins, small and large delta
antigens (SDAg and LDAg), they might interact with hosts’ molecules
and make some symptoms. Our lab has established Caenorhabditis
elegans (C.elegans) as a platform to study whether HDV antigens will
affect worm growth. In previous studies, transgenic worms expressing
SDAg in pharynx caused growth retard. In my thesis, I used different
length of SDAgs named SD441, SD327, SD267, and SD201 to dissect
the domain of delta antigens, which leads to growth retard, as well the
effect of different genotypes (genotype I, genotype II, genotype III) and
expression at different stages. I found that (1) the influence are distinct in
all different length of SDAgs, except SD441, the SDAg without
nuclear-localization signal ( NLS ), (2) SDAg expression at L4 stage has
the most serious effect, and (3) the degree of effect is genotype II >
genotype I > genotype III. These results show that the entrance of SDAg
into hosts’ nucleus is very important to result in growth retardation and
egg-laying activity. SDAg might interfere with unknown molecules in the
nucleus. Therefore, if these molecules would be identified, we could
search for the homologous genes in human, and possible to delineate the
molecular mechanisms involved in the pathogenesis of HDV.

誌謝.............................................................................................................i
中文摘要....................................................................................................ii
Abstract.....................................................................................................iii
目錄...........................................................................................................iv
前言............................................................................................................1
實驗目的..................................................................................................10
材料與方法..............................................................................................11
實驗結果..................................................................................................25
討論..........................................................................................................34
參考文獻..................................................................................................39
圖..............................................................................................................46
附錄..........................................................................................................56

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