臺灣博碩士論文加值系統

全身性紅斑狼瘡 (Systemic Lupus Erythematosus， SLE) 是屬於典
型的全身性自體免疫疾病，其特色是在 SLE 病人血清中產生針對抗核抗
體。 DNase 是一種核酸內切，其主要的功能在於能夠將去氧核糖核
酸 (DNA) 水解，在正常人類的血液及尿液中此都存在。 在我們的研
究中顯示當利用酵素免疫分析的方式檢測 anti-DNase antibodies大約
有 62.4% 的SLE 病人是呈現陽性反應﹔而在抗核抗體為陰性的對照組
血清中僅 8% 為陽性反應。 另外 SLE 病人血清中 anti-DNase
antibodies 與 anti-DNA antibodies 是呈現正相關。 我們也收集
SLE 病人的血清經由親和性管柱純化的方式純化 anti-DNase IgG，
此種抗體經由酵素免疫法測試可以結合至 DNase 及 DNA﹔若將合成的
DNase peptide 與牛胰臟 DNase I 進行競爭性試驗則可以抑制與
anti-DNase IgG 和牛的 DNase I 結合。 當我們利用親和性管柱所
純化的 anti-DNase IgG 做間接免疫螢光分析法時，發現以 Hep-2
細胞作為受質時會呈現斑點型及核仁型的分佈，但不會在鞭毛蟲
(Crithidia luciliae) 作為受質時不會和其 kinetoplast 結合。
進一步利用親和性管柱所純化的 anti-DNase IgG 比正常人血清所
純化的 IgG 更明顯抑制鏈球菌、牛、人的 DNase 活性。 值得注意
的是，若利用牛的 DNase I 去免疫 BALB/c 及 NZB/NZW 老鼠所產生
的 anti-DNase antibodies 及 anti-DNA antibodies 的效價都較正
常之未被免疫老鼠來的高。此外，取用牛的 DNase I 免疫 BALB/c老
鼠的腎臟做組織學切片，發現在老鼠腎管壁有纖維細胞的替代。
綜合以上結果我們認為，anti-DNase antibodies 普遍存在於 SLE
病人血清中，並且經由親和性管柱所純化的此種 anti-DNase
antibodies 可以抑制 DNase的活性。 因此推測在 SLE 病人血清中，
可能由於 anti-DNase antibodies 的存在而抑制 DNase 水解 DNA
的活性，進而刺激或促進 anti-DNA antibodies的產生。

英文摘要
Systemic lupus erythematosus (SLE) is a prototypic systemic
autoimmune disease characterized by the production of
autoantibodies against nuclear components . Dexoxyribonuclease I
(DNase I), an endonuclease capable of hydrolyzing of DNA, is
normally present in human serum and urine. In our study,
antibodies binding to bovine DNase I were investigated by ELISA
in SLE patients and anti-nuclear antibody (ANA) negative
normal sera. About 62.4% of SLE patients (63/101) were positive
for anti-DNase antibodies while only 8% of normal controls
(8/98) were positive. A positive correlation was found between
the concentrations of anti-DNase and anti-DNA antibodies in
SLE patients. Affinity-purified anti-DNase IgG from pooled SLE
patients serum and BALB/c mice immunized with bovine DNase
bound to DNase as well as DNA coated ELISA plates. A synthetic
peptide which cross-response to the catalytic site of DNase was
able to completely inhibit anti-DNase IgG bound to DNase but
not DNA coated plate. Indirect immunofluorescence of anti-DNase
IgG displayed speckled and nucleolar patterns of Hep-2 cells
but no reaction to the kinetoplast DNA in Crithidia luciliae.
In addition to bovine DNase, anti-DNase IgG also bound to DNase
in streptococcal supernatants and human urine and inhibited
their enzymatic activities . It was also noticed that BALB/c
and NZB/NZW mice immunized with bovine DNase I produced more
anti-DNase and anti-DNA antibodies than na鴳e mice. Besides,
immunized BALB/c mice showed fibroblast cells in renal tubule
walls under histological examination showed.Based on the
data above, we conclude that it is possible that the
inhibition of DNase I activity by the anti-DNase antibodies
may cause a defect in DNA degradation and stimulate or
enhance anti-DNA antibodies production.

目錄
中文摘要
英文摘要
誌謝
縮寫
圖目錄
前言
實驗設計
材料與方法
一、材料與方法
I. 人類血清
II. 實驗動物
III. Peptides的合成
IV. 酵素免疫
V. 親和性管柱層析法
VI. 測定抗體濃度
VII. 間接免疫螢光染色法
VIII. 硫酸十二酯鈉-聚丙烯醯氨膠電泳法
IX. 西方點墨法
X. 酵素活性膠體試驗
XI. 抗體抑制 DNase 活性試驗
XII. 免疫小鼠
XIII腎臟組織學切片
二、試藥與儀器
結果
I. 檢測 SLE 病人血清中的 anti-DNase antibodies
II. Anti-DNase IgG的特性
III. Anti-DNase antibodies對 NZB/NZW F1及
BABL/c老鼠的影響
討論
圖
參考文獻
自述
著作權聲明

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