摘要
利用粒子作為藥物遞送系統可以增強多種免疫反應，而此增益效果使其具有用於疫苗載體的潛力。在近期的研究中，研究人員發現顆粒的大小和形狀會影響它們與免疫系統的相互作用，但免疫系統如何區分顆粒結構的潛在機制尚未明朗。為了探討這個問題，在本研究中，我們合成了三種不同形狀（spherical, spiky, and spanish-needle）的鋁金屬有機骨架（Al-MOF），此金屬有機框架為一可同時作為免疫佐劑和攜帶抗原(卵清蛋白, OVA)的次單元疫苗，同時我們也會探討三種不同型態的Al-MOF疫苗對於刺激並延長抗原特異性免疫反應的能力。MOFs為金屬離子和有機配位體所構成的多孔材料，由於其卓越的設計靈活性和生物降解性，MOF已在各種生物醫學應用中被用作載體系統以遞送核酸或蛋白質藥物；鋁鹽為常用的免疫佐劑，具有優異的生物相容性以及能有效增強抗原所引起的免疫反應；OVA則是已被廣泛用於免疫研究中的模型抗原。在材料測試方面，我們通過掃描式電子顯微鏡(SEM)、能量色散X射線(EDX)光譜、動態光散射(DLS)和粉末 X 射線衍射儀(PXRD)觀察這三種Al-MOF的形態和特徵，並成功將OVA 加載至此載體中。在細胞實驗方面，Al-MOF的濃度即使達到300 μg/mL，仍然沒有觀測到顯著的細胞毒性。同時，我們也針對巨噬細胞對不同形狀MOF的細胞攝取及其後續的免疫刺激反應進行研究。在未來的研究中，我們擬經由皮下注射的方式將此載體打入小鼠體內，並評估此載體系統的長期抗原特異性免疫反應。

關鍵詞：疫苗， 免疫原性， 金屬有機骨架， 卵清蛋白 (OVA) ， 抗原

Abstract
Particles-associated drug delivery systems are promising for vaccination as they can enhance diverse immune responses. Research has shown that the size and shape of particles could affect their interaction with immune systems. However, the underlying mechanism by which the immune system distinguishes the structure of antigen-presenting particulates has yet to be fully understood. To address this issue, three different shapes (spherical, spiky, and Spanish-needle) of the aluminum-based metal-organic framework (Al-MOF) can act synergistically as an adjuvant and a delivery vehicle is synthesized to carry model antigen ovalbumin (OVA), serving as a vaccine. The ability of these three Al-MOF vaccines in antigen presentation and following processing by the immune cells to elicit a potent and prolonged antigen-specific response was accessed. MOFs are a class of porous hybrid materials constructed from metal ions and organic linkers. Owing to their superior design flexibility and intrinsic biodegradability, MOFs have been used as carrier systems to deliver nucleic acids or protein drugs in various biomedical applications. Aluminum salts are frequently used adjuvants due to their excellent biocompatibility and their ability to boost immune responses to various antigens. OVA has been widely used as a model vaccine in immunization studies. The morphology and characteristics of these three Al-MOFs were confirmed by scanning electron microscopy (SEM), energy-dispersive X-ray (EDX) spectroscopy, dynamic light scattering (DLS), and powder X-ray diffractometer (PXRD). In addition, OVA could be successfully loaded into Al-MOFs. In vitro results showed that no significant cytotoxicity of these three Al-MOFs was detected at concentrations of up to 300 μg/mL. The cell uptake of different shapes of MOFs by macrophages and their ability to activate the subsequent immune responses will be studied. In future works, the subcutaneous tissue of test mice will be established and the prolonged antigen-specific response of such MOFs system will be evaluated.

Keywords: vaccine, immunogenicity, metal-organic frameworks, ovalbumin (OVA), antigen

Table of Contents
摘要 i
Abstract ii
Contents iii
List of Figures v
List of Table vi
List of Abbreviation vii
Chapter 1: Introduction 1
1.1 Subunit Vaccine 1
1.2 Pollen 1
1.3 Research Design 2
Chapter 2: Results and Discussion 4
2.1 Characteristic of Al-MOFs 4
2.2 Cytotoxicity tests 9
2.3 Uptake of Al-MOFs by macrophages 9
2.4 Characteristics of OVA@MOFs 11
2.5 Uptake of OVA@MOFs by macrophages 13
2.6 In vitro degradation of OVA@MOFs 14
2.7 In vitro immune activation of OVA@MOFs 15
2.8 Conclusion 16
Chapter 3: Materials and Methods 17
3.1 Experimental Materials 17
3.2 Preparation and Characteristics of Al-MOFs 17
3.3 Preparation and Characterization of OVA@MOFs 17
3.4 Cell Viability Assay 18
3.5 Activation and Uptake of MOFs to Macrophages 18
References 20

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