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標題: | 氧化鐵奈米粒子對小鼠初代微膠細胞處理及呈現抗原的影響 The Effect of Iron Oxide Nanoparticles on the Antigen Processing and Presenting in Murine Microglia |
作者: | Chih-hua Shih 施芷華 |
指導教授: | 詹東榮(Tong-Rong Jan) |
關鍵字: | 氧化鐵奈米粒子,微膠細胞,抗原吞噬,抗原處理,抗原呈現,共同刺激分子,介白素, antigen uptake,antigen processing,antigen presenting,costimulatory molecule,interleukin,iron oxide nanoparticles,microglia,murine, |
出版年 : | 2013 |
學位: | 碩士 |
摘要: | 氧化鐵奈米粒子廣泛應用於生物醫學及臨床上核磁共振成像的影像對比劑,包括了中樞神經系統的造影。微膠細胞為中樞神經系統中類似巨噬細胞的免疫細胞,在神經免疫扮演重要功能,且負責處理入侵中樞的外來抗原。本研究目的在於探討氧化鐵奈米粒子對小鼠微膠細胞攝入、處理及呈現抗原的影響。初代小鼠微膠細胞暴露於氧化鐵奈米粒子後,以流式細胞儀藉由pHrodo E. coli 粒子, Alexa647-卵白蛋白 (OVA)以及DQ-OVA分別分析其抗原吞噬、攝入及處理的能力。而抗原呈現的能力則是在脂多醣及IFN-r刺激後,以流式細胞儀及real-time RT-PCR測定其相關因子,包括了MHCII,共同刺激分子CD40、CD80、CD86以及介白素-12、23、27的表現。此外,氧化鐵奈米粒子對微膠細胞刺激T淋巴球的能力則以與抗原免疫過小鼠之T淋巴球共同培養後T淋巴球所表現的細胞激素進行評估。實驗結果顯示氧化鐵奈米粒子的暴露會抑制微膠細胞的吞噬作用,及對OVA的攝入和處理能力。且氧化鐵奈米粒子會抑制CD40及IL-12 p35 mRNA的表現,但促進IL-23、CD80及CD86 mRNA以及細胞膜上CD86的表現。另外,氧化鐵奈米粒子處理後的微膠細胞對T淋巴球刺激的能力可藉由IL-2及IFN-r的分泌上升可見顯著的增加。綜合上述,本研究結果指出氧化鐵奈米粒子雖抑制了微膠細胞吞噬及處理抗原的能力,卻促進了微膠細胞刺激T 淋巴球的能力,並伴隨共同刺激分子及介白素的改變。 Superparamagnetic iron oxide nanoparticles have been employed as magnetic resonance imaging (MRI) contrast agents for a variety of biomedical research and clinical applications, including the imaging of the central nervous system (CNS). As the central resident immune cells with macrophage-like functions, microglial cells play a key role in neuroimmunity and are the dominant cells responsible for managing foreign materials entering to the CNS. The objective of this study was to investigate the effect of iron oxide nanoparticles on the antigen-uptake, processing and presenting activities of murine microglia. Primary murine microglial cells were exposed to iron oxide nanoparticles, and their phagocytotic, antigen uptake and processing activities were determined by flow cytometry using pHrodo E. coli bioparticles, Alexa FluorR 647 conjugated with ovalbumin (Alexa647-OVA) and DQ-OVA, respectively. The expression of antigen presentation-related molecules, including major histocompatibility complex (MHC) II, the costimulatory molecules CD40, CD80, and CD86 and the cytokines IL-12, IL-23 and IL-27 by microglia stimulated with lipopolysaccharide (LPS) and interferon-r was measured by flow cytometry and real time RT-PCR. In addition, the capability of iron oxide nanoparticle-exposed microglia to stimulate T cell cytokine production was evaluated by co-culture of microglia and antigen-primed splenocytes. Exposure to iron oxide nanoparticles attenuated the phagocytosis of pHrodo E. coli and the uptake and processing of ovalbumin (OVA) by microglia. Iron oxide nanoparticles decreased the CD40 and IL-12 p35 mRNA expression, but increased the expression of CD80, CD86 and IL-23 mRNA and the cell membrane CD86. In addition, the capacity of microglia to stimulate OVA-primed splenocytes was enhanced by iron oxide nanoparticles, as evidenced by the increased production of IL-2 and IFN-r. Collectively, the present study demonstrated that iron oxide nanoparticles affected microglial functionality. The phagocytic, antigen-uptake and processing activities were attenuated. In contrast, the capability of microglia to stimulate T cells was enhanced, which may be attributed to the alteration of the expression of surface markers and cytokines. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/60226 |
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顯示於系所單位: | 獸醫學系 |
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