本文選題：微小隱孢子蟲 + IL-17�。� 參考：《西北農(nóng)林科技大學(xué)》2015年碩士論文

【摘要】：微小隱孢子蟲(Cryptosporidium parvum)是一種全球性分布的寄生于宿主消化道上皮細胞刷狀緣等的人獸共患寄生原蟲,嚴重威脅著人和家畜的健康。自1907年Tyzzer首先在小鼠的胃消化腺上皮細胞內(nèi)發(fā)現(xiàn)鼠隱孢子蟲(C.muris)以來,已在包括人在內(nèi)的5類脊椎動物體內(nèi)發(fā)現(xiàn)了隱孢子蟲屬的27個有效種。由C.parvum引起的隱孢子蟲病主要危害人和牛、山羊、綿羊等重要經(jīng)濟動物,尤其犢牛在動物和人的隱孢子蟲病互傳中扮演著主要角色。由于目前隱孢子蟲病防治尚缺乏有效的藥物和疫苗,因此,深入探討宿主對隱孢子蟲免疫防御機制、尤其是弄清宿主抗隱孢子蟲感染的關(guān)鍵細胞亞群及其作用機制,將為尋找關(guān)鍵靶標、研發(fā)預(yù)防性疫苗及特異性藥物和有效防控隱孢子蟲病提供思路。本研究首先對兩株Ⅱd亞型的C.parvum進行鑒定,選取合適的分離株感染BALB/c小鼠,建立感染模型;以感染模型為基礎(chǔ),檢測C.parvum感染BALB/c小鼠的脾臟、小腸相關(guān)淋巴組織中Th17主效應(yīng)細胞因子IL-17及Th17細胞分化、效應(yīng)相關(guān)細胞因子、轉(zhuǎn)錄因子及轉(zhuǎn)導(dǎo)分子在不同時間點的表達變化情況,以此來明確Th17細胞在C.parvum感染和清除中的作用。1.為建立合適的感染動物模型,將C.parvum的兩個分離株進行鑒定,并選取ⅡdA15G2R1亞型作為受試卵囊,按照每只1×106個微小隱孢子蟲卵囊分別經(jīng)口感染BALB/c小鼠,測定其排卵囊規(guī)律,發(fā)現(xiàn)小鼠于感染后1 d到20 d連續(xù)排卵囊,感染后21 d停止排卵囊;并對其排出卵囊進行形態(tài)學(xué)及基因型(18S rRNA)鑒定,遺傳穩(wěn)定性良好。2.應(yīng)用實時定量PCR(Quantitative Real time-PCR)測定感染后不同時間脾臟和腸相關(guān)淋巴組織中Th17細胞亞群分化相關(guān)促進因子(TGF-β、IL-6)、轉(zhuǎn)錄因子(RORγt)和信號傳導(dǎo)分子(STAT-3)的表達和變化,結(jié)果顯示在C.parvum感染和清除過程中,宿主Th17細胞亞群分化相關(guān)細胞因子都出現(xiàn)了不同程度的上調(diào)表達。3.應(yīng)用實時定量PCR方法測定感染后不同時間脾臟和腸相關(guān)淋巴組織中Th17細胞分化維持細胞因子(IL-23)、效應(yīng)細胞因子(IL-17、IL-22、TNF-α)等的表達和變化;并采用酶聯(lián)免疫吸附試驗(ELISA)檢測感染后小鼠組織IL-17的蛋白水平變化情況,分析Th17細胞亞群在C.parvum感染和清除過程中的效應(yīng)情況。主效應(yīng)因子IL-17在感染早期、中期都有較高水平的表達;其他效應(yīng)因子幾乎在整個感染過程中都有較高水平的表達。
[Abstract]:Cryptosporidium parvum (Cryptosporidium parvum) is a globally distributed zoonotic parasite parasitic on the epithelial cells of the host digestive tract, which is a serious threat to the health of human and domestic animals. Since Tyzzer first found Cryptosporidium muris in mouse gastric digestive gland epithelium in 1907, 27 effective species of Cryptosporidium have been found in 5 kinds of vertebrates including human. Cryptosporidiosis caused by C. parvum mainly harms people and important economic animals such as cattle, goats and sheep, especially calves play a major role in the transmission of Cryptosporidiosis between animals and humans. Due to the lack of effective drugs and vaccines in the prevention and treatment of Cryptosporidiosis, the mechanism of host immune defense against Cryptosporidium is discussed, especially the key cell subsets of host against Cryptosporidium infection and its mechanism of action. It will provide ideas for finding key targets, developing prophylactic vaccines and specific drugs and effectively preventing and controlling Cryptosporidiosis. In this study, two strains of C.parvum with subtype 鈪，

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