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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 林建達 | zh_TW |
| dc.contributor.advisor | Jian-Da Lin | en |
| dc.contributor.author | 陳言亭 | zh_TW |
| dc.contributor.author | Yen-Ting Chen | en |
| dc.date.accessioned | 2023-10-03T16:11:36Z | - |
| dc.date.available | 2023-11-10 | - |
| dc.date.copyright | 2023-10-03 | - |
| dc.date.issued | 2023 | - |
| dc.date.submitted | 2023-08-04 | - |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/90463 | - |
| dc.description.abstract | 第一型免疫反應及慢性發炎與動脈粥狀硬化的進程息息相關，而近期的臨床目標在於透過增加第二型免疫反應以降低發炎症狀來促進斑塊消退。核苷酸結合寡聚域蛋白質2 (NOD2) 受體能偵測細菌或其產物以刺激免疫反應。根據先前研究指出，Nod2-/-小鼠有腸道菌相失衡及較少的第二型免疫反應等現象，然而腸道菌群失衡，動脈粥狀硬化疾病進程及第一/二型免疫反應之間的相關機制仍待探究，因此本研究將探討腸道菌群失調是否能促進動脈粥狀硬化的進展，而增加第二型免疫反應是否能治療疾病。本研究預期透過鼠鞭蟲 (T. muris) 及多型性螺旋線蟲 (H. polygyrus) 腸道寄生蟲的感染能抑制發炎反應使斑塊消退。我們建立了高參數免疫分析圖譜，透過光譜式流式細胞分析技術可同時分析組織中35種免疫細胞標記，並在疾病進展及逆轉期間長期對野生型 (WT) 及Nod2-/-小鼠的血液進行免疫細胞群分析。結果顯示，淋巴細胞在兩種基因型小鼠中的表現量有顯著差異，在疾病進程中Ly6C+/Ly6C-單核球、樹突細胞、γ/δ T細胞及effector memory CD4/CD8 T細胞顯著增加，而CD25+ CD4 T細胞則降低其表現。此外，經過寄生蟲感染後的動脈粥狀硬化小鼠發現腹腔組織中的M2巨噬細胞、CD25+ ST2+調節性T細胞、先天性淋巴細胞2及活化的CD4/CD8 T細胞群擴增。最後，透過切片定量斑塊大小，我們成功利用寄生蟲感染誘導動脈粥狀硬化小鼠主動脈弓的斑塊消退，且證實了斑塊消退依賴於NOD2基因的存在。因此，由本實驗結果顯示，基於強烈第二型免疫反應的生成，寄生蟲感染將成為未來治療動脈粥狀硬化的潛在策略。 | zh_TW |
| dc.description.abstract | Type 1 immune responses and chronic inflammation are always associated with atherosclerosis progression. A recent clinical goal aims to promote plaque regression by reducing inflammation that may be correlated with the increase of type 2 immunity. Nucleotide-binding oligomerization domain-containing protein 2 (NOD2) is the bacterial sensor to stimulate immune responses, and previous studies have indicated Nod2-/- mice present gut dysbiosis and reduced type 2 immunity. However, the relationships between gut dysbiosis, atherosclerosis progression, and type 1/2 immune responses remain elusive. Here, we aim to interrogate whether gut dysbiosis can promote atherosclerosis progression and whether the inflammatory phenotypes can be reversed by increasing type 2 immunity. We hypothesized that the gastrointestinal helminth infection, Trichuris muris (T. muris) and Heligmosomoides polygyrus (H. polygyrus), can resolve inflammation and induce plaque regression. Currently, the powerful immune profiling panel is built up to analyze the 35 immune markers simultaneously by spectral flow cytometry. We longitudinally perform immune profiling on the blood of wild-type (WT) and Nod2-/- mice during atherosclerosis progression and regression. The significant differences of B cells and CD4 T cells were observed in both genotypes of mice. We show that Ly6C+/Ly6C- monocytes, dendritic cells, γ/δ T cells, and effector memory CD4/CD8 T cells increase in the blood, while CD25+ CD4 T cells decrease during atherogenesis progression. Notably, strong type 2 immune responses were induced by helminth-infected atherosclerotic mice and may induce plaque regression, as evidenced by the expansion of M2 macrophages, CD25+ ST2+ regulatory T cells, type 2 innate lymphoid cells, and activated CD4/CD8 T cells in peritoneal cavity tissue. Plaque regression in the aortic root of atherosclerotic mice was observed with helminth treatments, as confirmed through quantitative analysis. Additionally, our study reveals that plaque regression is NOD2-dependent. Overall, our results suggest promising strategies by helminth treatments for future therapeutics in atherosclerosis. | en |
| dc.description.provenance | Submitted by admin ntu (admin@lib.ntu.edu.tw) on 2023-10-03T16:11:36Z No. of bitstreams: 0 | en |
| dc.description.provenance | Made available in DSpace on 2023-10-03T16:11:36Z (GMT). No. of bitstreams: 0 | en |
| dc.description.tableofcontents | 口試委員會審定書 I
Acknowledgments II Abstract IV 中文摘要 VI Abbreviation VII Contents XI List of Figures XV Chapter 1 Introduction 1 1.1 Atherosclerosis 1 1.2 The critical role of monocyte-macrophages in atherosclerosis 6 1.3 The importance of nucleotide-binding oligomerization domain (NOD)-like receptors (NLRs) to maintain gut homeostasis by preventing chronic inflammation 12 1.4 The relationship between type 1/2 immunity and plaque progression/regression 16 1.5 The interplay of helminth infection and type 2 immunity in the gut 20 1.5.1 Trichuris muris 24 1.5.2 Heligmosomoides polygyrus 27 1.6 Aims and Objectives of the study 31 Chapter 2 Materials and Methods 33 2.1 Experimental models and Subjects details 33 2.1.1 Mice 33 2.1.2 Experimental design and sample collection 33 2.2 Polymerase chain reaction (PCR) genotyping of Nod2-/- mice 36 2.2.1 Genomic DNA making from mice 36 2.2.2 The making of primer Stock (100 µM) and working concentration (10 µM) of primers 36 2.2.3 PCR reactions 37 2.2.4 Electrophoresis 37 2.3 Total cholesterol assays 38 2.4 Parasite cultivation, infection and counting 39 2.4.1 Trichuris muris 39 2.4.1.1 Cultivation 39 2.4.1.2 Infection 41 2.4.1.3 Adult worms count 41 2.4.2 Heligmosomoides polygyrus 42 2.4.2.1 Cultivation 42 2.4.2.2 Infection 44 2.4.2.3 Adult worms count 44 2.4.3 Fecal eggs enumeration 45 2.5 Flow cytometry 46 2.5.1 Sample preparation 46 2.5.1.1 Blood collection 46 2.5.1.2 Isolation of Peritoneal cavity cells (PEC) 47 2.5.1.3 Separation of aorta-bound cells 48 2.5.2 Flow cytometry staining 50 2.5.2.1 35 immune markers 50 2.5.2.2 Nuclear protein (Th panel) 51 2.5.3 Compensation (reference tubes) preparation 53 2.5.4 Flow cytometry analysis 53 2.5.5 Data analysis 54 2.6 Immunohistochemistry (IHC) Staining 54 2.6.1 Arch frozen section 54 2.6.2 CD68 staining 54 2.6.3 Aortic Root Imaging 56 2.7 Principal component analysis (PCA) 57 2.8 Quantification and statistical analysis 57 Chapter 3 Results 58 3.1 Immune profiling panel between different tissues 58 3.2 The impact of different conditions with Trichuris muris eggs infection for inducing stronger type 2 immunity in mice 60 3.3 AAVmPCSK9 injection induces hypercholesterolemia and high body weight in mice 64 3.4 Helminth infection with chow diet switch can reverse hypercholesterolemia and reduce body weight induced by AAVmPCSK9 injection 66 3.5 Significant differences in lymphocytes expression were identified in blood between wild-type and Nod2-/- mice during atherosclerosis progression 67 3.6 Helminth infection induces an increase in specific immune cell populations during atherosclerosis regression 71 3.7 Helminth infection induces a strong type 2 immune response during atherosclerosis regression 84 3.8 Helminth infections can induce regression of atherosclerosis in rAAVmPCSK9 injected mice 92 Chapter 4 Discussion 138 4.1 Interplay of helminth infection, NOD2 gene, and immune responses in atherosclerosis: Unforeseen findings and implications 138 4.2 Gut dysbiosis, metabolite imbalance, and their role in atherosclerosis development: Insights from Nod2-/- mice 139 4.3 NOD2 and atherosclerosis: Deciphering contradictions and pursuing mechanistic insights 141 4.4 Modulation of immune cell populations in atherosclerosis: Impact of NOD2 deficiency and helminth infections 143 4.5 Critical role of T and B Cell activation markers in atherosclerosis progression and regression: Insights from immune profiling 145 4.6 Further studies and limitations: The impact of diet, infection, and immune responses on atherosclerosis 148 4.7 Limitations of current mouse models and role of T cells in atherosclerosis research 150 4.8 Emerging therapies for atherosclerosis and the potential role of helminth infection, E/S proteins, and gut microbiota in disease regression 151 4.9 Future therapies: Targeting microbiota for atherosclerosis treatment 155 4.10 Conclusions: Unraveling therapeutic potential in atherosclerosis 157 References 159 Appendix 176 | - |
| dc.language.iso | en | - |
| dc.subject | 寄生蟲感染 | zh_TW |
| dc.subject | 動脈粥狀硬化 | zh_TW |
| dc.subject | NOD2 基因 | zh_TW |
| dc.subject | 鼠鞭蟲 | zh_TW |
| dc.subject | 多型性螺旋線蟲 | zh_TW |
| dc.subject | 第二型免疫反應 | zh_TW |
| dc.subject | Heligmosomoides polygyrus | en |
| dc.subject | Nod2-/- | en |
| dc.subject | Trichuris muris | en |
| dc.subject | atherosclerosis | en |
| dc.subject | helminth infection | en |
| dc.subject | type 2 immunity | en |
| dc.title | 治療動脈粥狀硬化的未來展望：寄生蟲感染及第二型免疫反應 | zh_TW |
| dc.title | Possible future therapeutic for atherosclerosis: Helminth infection and type 2 immunity | en |
| dc.type | Thesis | - |
| dc.date.schoolyear | 111-2 | - |
| dc.description.degree | 碩士 | - |
| dc.contributor.oralexamcommittee | 林甫容;楊鎧鍵;徐嘉琳 | zh_TW |
| dc.contributor.oralexamcommittee | Fu-Jung Lin;Kai-Chien Yang;Chia-Lin Hsu | en |
| dc.subject.keyword | 動脈粥狀硬化,NOD2 基因,鼠鞭蟲,多型性螺旋線蟲,第二型免疫反應,寄生蟲感染, | zh_TW |
| dc.subject.keyword | atherosclerosis,Nod2-/-,Trichuris muris,Heligmosomoides polygyrus,type 2 immunity,helminth infection, | en |
| dc.relation.page | 202 | - |
| dc.identifier.doi | 10.6342/NTU202303056 | - |
| dc.rights.note | 同意授權(限校園內公開) | - |
| dc.date.accepted | 2023-08-08 | - |
| dc.contributor.author-college | 生命科學院 | - |
| dc.contributor.author-dept | 生化科技學系 | - |
| dc.date.embargo-lift | 2028-09-01 | - |
| 顯示於系所單位： | 生化科技學系 | |
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