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| ???org.dspace.app.webui.jsptag.ItemTag.dcfield??? | Value | Language |
|---|---|---|
| dc.contributor.advisor | 陳慧文 | zh_TW |
| dc.contributor.advisor | Hui-Wen Chen | en |
| dc.contributor.author | 甘曜銘 | zh_TW |
| dc.contributor.author | Yao-Ming Kan | en |
| dc.date.accessioned | 2023-08-09T16:08:23Z | - |
| dc.date.available | 2023-11-10 | - |
| dc.date.copyright | 2023-08-09 | - |
| dc.date.issued | 2023 | - |
| dc.date.submitted | 2023-07-21 | - |
| dc.identifier.citation | 陳耀云, 2022. 四種不同型別冠狀病毒核殼蛋白的交叉抗原性探討. 國立臺灣大學, 台北市.
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Both Feline Coronavirus Serotypes 1 and 2 Infected Domestic Cats Develop Cross-Reactive Antibodies to SARS-CoV-2 Receptor Binding Domain: Its Implication to Pan-CoV Vaccine Development. Viruses 15. Yang, D.-K., Park, Y.-R., Kim, E.-j., Lee, H.J., Shin, K.-S., Kim, J.-H., Lee, K., Hyun, B.-H., 2022. Incidence and sero-surveillance of feline viruses in Korean cats residing in Gyeonggi-do. Korean Journal of Veterinary Research 62, e24. Yen, S.J., Chen, H.W., 2021. Feline Coronaviruses Identified in Feline Effusions in Suspected Cases of Feline Infectious Peritonitis. Microorganisms 9. Yin, Y., Li, T., Wang, C., Liu, X., Ouyang, H., Ji, W., Liu, J., Liao, X., Li, J., Hu, C., 2021. A retrospective study of clinical and laboratory features and treatment on cats highly suspected of feline infectious peritonitis in Wuhan, China. Sci Rep 11, 5208. Yuki, M., Aoyama, R., Nakagawa, M., Hirano, T., Naitoh, E., Kainuma, D., 2020. A Clinical Investigation on Serum Amyloid A Concentration in Client-Owned Healthy and Diseased Cats in a Primary Care Animal Hospital. Vet Sci 7. Ziebuhr, J., 2005. The coronavirus replicase. Curr Top Microbiol Immunol 287, 57-94. Ziebuhr, J., Snijder, E.J., Gorbalenya, A.E., 2000. Virus-encoded proteinases and proteolytic processing in the Nidovirales. J Gen Virol 81, 853-879. | - |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/88235 | - |
| dc.description.abstract | 貓傳染性腹膜炎 (FIP) 是由貓冠狀病毒 (FCoV) 其中一種生物型，貓傳染性腹膜炎病毒 (FIPV) ，引起之貓致命性疾病。而對於FIPV感染期間的病理免疫機制，尤其是抗體依賴增強作用 (ADE) 缺乏研究，這導致該病毒疫苗開發方面存在不確定因素。此外，貓對於近年造成全球威脅的第二型嚴重急性呼吸道綜合症冠狀病毒 (SARS-CoV-2) 也具有感受性，當貓同時感染這兩種冠狀病毒，FIPV和SARS-CoV-2之間的潛在交叉反應便成了一大議題。本研究旨在通過對FIP患貓的積液樣本中的病毒和抗體進行分析，以增加我們對FIP病理發生機制和冠狀病毒交叉反應的了解。從2022年1月開始，46個疑似FIP貓腹水樣本經PCR和免疫螢光試驗 (IFA) 的重複驗證確診為陽性。而首先，我們對這些陽性案例的特徵進行分析以揭示FIP的危險因子，發現了2歲以下以及白蛋白與球蛋白比值 (A/G ratio) 小於等於0.4的貓隻與FIP陽性有著顯著關聯。隨後，我們成功地在36個陽性FIP積液樣本中進行了FCoV片段的序列分析，其中36個屬於基因一型，僅有一個檢體同時存在基因一型和二型FCoV，相對於基因二型FCoV，基因一型FCoV親緣樹狀圖顯示出高度多樣性。再者，積液當中的細胞激素TNF-α及IL-6濃度，雖在FIP陽性與陰性病例之間並無顯著差異，但仍使我們更進一步瞭解該疾病的免疫機制。此外，通過從FIP患貓積液中純化之免疫球蛋白G (IgG) 和建立的體外FIPV ADE測試模型，檢測到了FIP陽性積液樣本中的ADE現象。另一項研究探討了FCoV和SARS-CoV-2之間的潛在交叉反應，結果顯示，來自兔隻抗SARS-CoV-2核殼蛋白 (Nucleocapsid) 的多株抗體 (pAb) 通過西方墨點法 (Western blot) 和酵素連結免疫吸附試驗（ELISA）與FCoV產生反應，但在FIPV ADE測試模型沒有觀察到ADE現象。為進一步研究這兩種病毒之間的關係，將人類COVID-19康復血清引入測試，發現兩名患者的血清與FCoV的刺突蛋白 (Spike) 亞單位1 (subunit 1) 和核殼蛋白產生交叉反應。同時，一人的血清樣本甚至略微增強FIPV的感染狀態，是為ADE現象，也初步證實了兩種病毒的交叉反應。此外，將FIP貓積液中的IgG與SARS-CoV-2的刺突蛋白、核殼蛋白及其片段進行測試，發現IgG能夠與SARS-CoV-2的刺突蛋白亞單位2 (subunit 2) 和核膜蛋白的所有片段結合，這再次證明了FCoV和SARS-CoV-2之間的雙向交叉反應。本研究首次報告了FIP貓積液中IgG引發的ADE現象，並發現FCoV和SARS-CoV-2之間的潛在交叉反應，為未來FIPV之感染研究提供了更近一步的見解。 | zh_TW |
| dc.description.abstract | Feline infectious peritonitis (FIP) is a fatal disease in cats caused by feline infectious peritonitis virus (FIPV), a biotype of feline coronavirus (FCoV). The lack of research on antibody-dependent enhancement (ADE) during FIPV pathogenesis has resulted in uncertainties in vaccine development. Moreover, cats are susceptible to severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), another coronavirus posing a pandemic threat, and the potential cross-reactivity between these two coronaviruses remains unclear. This study aimed to increase our knowledge of FIP pathogenesis and coronavirus cross-reactivity by characterizing the virus and antibodies in FIP-diseased effusion samples. From January 2022, 46 FIP-confirmed samples were collected using PCR and IFA simultaneously. The signalments of these cases were firstly analyzed to reveal the FIP risk factors, which discovered that the cats with age equal to or lower than two y/o and A/G ratio equal to or lower than 0.4 were significantly correlated to FIP-positive. Subsequently, materials related to FIP pathogenesis in FIP effusions were investigated. FCoV fragments were successfully sequenced in 36 positive samples, with 35 cases belonging to genotype I and only one co-infected with genotype I and II FCoV. The phylogenetic analysis of the type I FCoV sequences revealed high diversity. Although there was no significant difference in the level of TNF-α and IL-6 in the effusions between FIP-positive and FIP-negative cases, this finding contributes to a better understanding of the immune mechanisms underlying FIP. The ADE phenomenon was also detected in FIP-positive effusion samples through IgG purified from FIP cat’s effusions and the established in vitro FIPV ADE test model. Another investigation explored the potential cross-reactivity between FCoV and SARS-CoV-2. Results revealed that only rabbit-derived anti-SARS-CoV-2 nucleocapsid pAb reacted with FCoV through western blot and ELISA, but no ADE phenomenon was observed. To further study the relationship between these two viruses, human COVID-19 convalescent serum was introduced into the test, which surprisingly demonstrated that serum from two patients cross-reacted with FCoV spike subunit 1 (S1) and nucleocapsid proteins—meanwhile, one human serum sample even slightly enhanced FIPV infection. Also, the IgG from FIP cat effusions was incubated with SARS-CoV-2 spike, nucleocapsid proteins, and their truncated fragments. The IgG was found capable of binding to the spike subunit 2 (S2) and all fragments of nucleocapsid protein of SARS-CoV-2, which proved the two-way cross-reactivity between FCoV and SARS-CoV-2. This study is the first to report the in vitro ADE phenomenon elicited from IgG in FIP effusions and discovered potential cross-reactivity between FCoV and SARS-CoV-2, providing valuable insights for future FCoV studies. | en |
| dc.description.provenance | Submitted by admin ntu (admin@lib.ntu.edu.tw) on 2023-08-09T16:08:23Z No. of bitstreams: 0 | en |
| dc.description.provenance | Made available in DSpace on 2023-08-09T16:08:23Z (GMT). No. of bitstreams: 0 | en |
| dc.description.tableofcontents | 中文摘要 I
Abstract III List of Figures X List of Tables XII Chapter 1 Introduction 1 1.1 Feline coronavirus 1 1.1.1 The structure of Feline Coronavirus 1 1.1.2 The genome of Feline Coronavirus 3 1.1.3 Genotypes of Feline Coronavirus 4 1.1.4 Biotypes of Feline coronavirus 5 1.1.5 Epidemiology and phylogenetic analysis of feline infectious peritonitis virus infection in Taiwan and other countries 6 1.2 Feline infectious peritonitis 8 1.2.1 Characteristics of feline infectious peritonitis 8 1.2.2 Clinical signs of feline infectious peritonitis 9 1.2.3 Diagnosis of feline infectious peritonitis 10 1.2.3.1 Overview 10 1.2.3.2 Diagnosis of FIPV infection from blood 11 1.2.3.3 Diagnosis of FIPV infection from effusions 13 1.2.4 Host immune response during feline infectious peritonitis 15 1.3 Antibody-dependent enhancement 17 1.4 SARS-CoV-2 and its relationship with cats 19 1.5 Cross-reactivity between SARS-CoV-2 and FIPV 20 1.6 Aim of this study 21 Chapter 2 Materials and Methods 23 2.1 Clinical feline effusion samples collection and processing 23 2.2 Indirect immunofluorescence assay 25 2.3 Nucleic acid extraction and RT-PCR 27 2.4 FCoV genotyping, sequencing, and phylogenetic analysis 28 2.5 Enzyme-linked immunosorbent assay (ELISA) 30 2.6 Western blot (WB) 32 2.7 Feline cytokine levels measurement 34 2.8 Feline effusion IgG purification 36 2.9 Cell and virus cultures 38 2.10 Viral titer determination and plaque assay 39 2.11 FIPV ADE assay and neutralizing test of purified IgG from FIPV-infected cats’ effusions 40 2.12 Antigenic analysis of anti-FCoV S and N protein IgG in each purified IgG sample. 41 2.13 Statistical analysis 42 Chapter 3 Results 43 3.1 Sample collection and experimental design 43 3.2 FIP cases confirmed by IFA and RT-PCR 43 3.3 Characteristics of FIP cases 44 3.4 FIPV genotyping and phylogenetic analysis 45 3.4.1 The proportion of genotype I and genotype II FIPV collected in this study 45 3.4.2 Phylogenetic analysis of genotype I FIPV sequences 46 3.4.3 Phylogenetic analysis of genotype II FIPV sequences 47 3.5 Cytokine profiling of FIPV positive and negative effusion samples 48 3.6 IgG purification: Sample selection, elution, desalting, and FCoV binding confirmation 49 3.6.1 IgG purification: the successful example 49 3.6.2 The examination of the binding ability of purified IgG against FCoV 50 3.7 FIPV ADE establishment and the assay with IgG from FIP cats’ clinical effusions 50 3.7.1 Successful FIPV ADE establishment 50 3.7.2 FIPV ADE assay with IgG purified from 4 FIP-positive cats and neutralizing test of these IgG samples 51 3.7.3 Antigenic analysis of cat effusion IgG 52 3.8 Cross-reactivity between FCoV and SARS-CoV-2 53 3.8.1 Cross-antigenicity test with FCoV whole virion and anti-SARS-CoV-2 spike S1 and S2, N, and M protein pAb 53 3.8.2 FIPV ADE assay with anti-SARS-CoV-2 spike S1 and S2, N, and M protein pAb 54 3.8.3 Cross-antigenicity test with FCoV whole virion and human COVID-19 convalescent serum 55 3.8.4 FIPV ADE assay with human COVID-19 convalescent serum 56 3.8.5 Two-way cross-antigenicity test with IgG purified from FIP-positive cat’s effusions and structural proteins from SARS-CoV-2 57 3.8.5.1 IgG purified from FIP-positive cat’s effusions and SARS-CoV-2 spike 57 3.8.5.2 IgG purified from FIP-positive cat’s effusions and SARS-CoV-2 nucleocapsid protein 58 Chapter 4 Discussion 60 Chapter 5 Figures and tables 71 Chapter 6 References 114 | - |
| dc.language.iso | en | - |
| dc.subject | 第二型嚴重急性呼吸道症候群冠狀病毒 | zh_TW |
| dc.subject | 貓傳染性腹膜炎 | zh_TW |
| dc.subject | 貓冠狀病毒 | zh_TW |
| dc.subject | 抗體依賴增強作用 | zh_TW |
| dc.subject | 交互作用 | zh_TW |
| dc.subject | Antibody-dependent enhancement | en |
| dc.subject | Feline coronavirus | en |
| dc.subject | Feline infectious peritonitis | en |
| dc.subject | SARS-CoV-2 | en |
| dc.subject | Cross-reactivity | en |
| dc.title | 分析傳染性腹膜炎貓隻積液中之貓冠狀病毒及抗體及其與第二型嚴重急性呼吸綜合症冠狀病毒之間的關聯性 | zh_TW |
| dc.title | Feline coronavirus and antibodies in effusions of cats with feline infectious peritonitis and their potential cross-reactivity with SARS-CoV-2 | en |
| dc.type | Thesis | - |
| dc.date.schoolyear | 111-2 | - |
| dc.description.degree | 碩士 | - |
| dc.contributor.oralexamcommittee | 張惠雯;張晏禎;林中惠 | zh_TW |
| dc.contributor.oralexamcommittee | Hui-Wen Chang;Yen-Chen Chang;CHUNG-HUI LIN | en |
| dc.subject.keyword | 貓傳染性腹膜炎,貓冠狀病毒,抗體依賴增強作用,交互作用,第二型嚴重急性呼吸道症候群冠狀病毒, | zh_TW |
| dc.subject.keyword | Feline infectious peritonitis,Feline coronavirus,Antibody-dependent enhancement,Cross-reactivity,SARS-CoV-2, | en |
| dc.relation.page | 125 | - |
| dc.identifier.doi | 10.6342/NTU202301829 | - |
| dc.rights.note | 同意授權(全球公開) | - |
| dc.date.accepted | 2023-07-24 | - |
| dc.contributor.author-college | 生物資源暨農學院 | - |
| dc.contributor.author-dept | 獸醫學系 | - |
| Appears in Collections: | 獸醫學系 | |
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| ntu-111-2.pdf | 17.3 MB | Adobe PDF | View/Open |
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