- 學位論文
痢疾阿米巴抗原誘導單核球源樹突細胞 基因表現之狀況:非HIV及HIV患者間之比較
Gene Expression Pattern of Monocyte Drived Dendritic Cell Stimulated by Entamoeba histolytica Antigen: Comparison of Non-HIV and HIV Candidates
摘要
痢疾阿米巴感染人體的腸道後造成的疾病除了阿米巴性大腸炎以外，尤以侵入性的阿米巴肝膿瘍(ALA)最常發生。根據台大醫院的統計，愛滋病毒(HIV)的帶原者發生肝膿瘍的比例約占全體ALA病患的59%，使得ALA有機會成為一個感染HIV的新指標。實驗中在HIV患者的T細胞受到抗原刺激後，由CD69表現量提升的幅度發現，10μg阿米巴的全細胞混合抗原的使用量對免疫反應的刺激強度最強。以HIV感染的有無配對著ALA發生的有無分出四種組別，分別收下各組受試者的周邊血液單核細胞，純化出單核球後，培養六天成為未成熟樹突細胞，加入阿米巴抗原後繼而再培養一天成為成熟樹突細胞。四組受試者以樹突細胞成熟前後來比對在基因晶片上表現量具有統計差異的基因。為找出HIV造成的影響，以有、無感染HIV的群組中發現感染HIV群組的239個基因及未感染HIV群組 30個基因具有統計意義而篩出。此兩群基因再分析找出與全體受試者有、無發生ALA具一致性的基因，分別找出14個基因及9個基因。受試者發生ALA下，不論是否感染HIV皆啟動了一個基因–sperm associated antigen 9，此基因與其他–感染HIV的13個及未感染HIV的8個基因群，都可能和容易發生ALA存有尚未研究出的關聯性。
並列摘要
In addition to amebic colitis, amebic liver abscess (ALA) is the most common invasive disease among the Entamoeba histolytica infection. According to the data from National Taiwan University Hospital, up to 59% of ALA cases had underlying HIV (human immunodeficiency virus) infection in recent 10 years. Thus the clinical identification of ALA may be a surrogate marker of HIV infection. In our titration experiments, we found 10 μg / mL of amoeba total lysate (amoeba antigen) was the optimal concentration for in vitro activation of cellular immune responses. We enrolled 7 subjects and divided them into 4 groups according to their HIV seropositivity and having ALA or not. We isolated their PBMC (peripheral blood mononuclear cells) and purified monocytes by negative isolation. These monocytes differentiated to immature dendritic cells (DC) during the 6 days of in vitro culture, and were stimulated by amoeba antigen for one day or not. We harvested these day 7 cells and analyzed their gene expression by genome-wide screening by DNA microarray technique. 4 groups of candidates is formed into clusters for analysis. In order to find out the influences of HIV infection, 239 genes of HIV cluster and 30 genes of non-HIV cluster are sifted with statistical meaning. Of all candidates, further analysis investigates consistency of gene expression with ALA production at all, 14 genes from HIV cluster and 9 genes from non-HIV cluster are found out via categorized difference of influence with HIV. A gene–sperm associated antigen 9 has appeared together of two cluster, this gene and other 13 genes from HIV cluster and 8 genes from non-HIV cluster are related to that has not worked out yet may be apt to present ALA.
參考文獻
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