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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
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dc.contributor.advisor | 陳培哲(Pei-Jer Chen) | |
dc.contributor.author | Chi-Hung Cheng | en |
dc.contributor.author | 鄭吉宏 | zh_TW |
dc.date.accessioned | 2021-05-14T17:43:01Z | - |
dc.date.available | 2017-09-25 | |
dc.date.available | 2021-05-14T17:43:01Z | - |
dc.date.copyright | 2015-09-25 | |
dc.date.issued | 2015 | |
dc.date.submitted | 2015-08-12 | |
dc.identifier.citation | Biswas, S. K., & Lopez-Collazo, E. (2009). Endotoxin tolerance: new mechanisms, molecules and clinical significance. Trends Immunol, 30(10), 475-487. doi: 10.1016/j.it.2009.07.009
Bleriot, C., Dupuis, T., Jouvion, G., Eberl, G., Disson, O., & Lecuit, M. (2015). Liver-resident macrophage necroptosis orchestrates type 1 microbicidal inflammation and type-2-mediated tissue repair during bacterial infection. Immunity, 42(1), 145-158. doi: 10.1016/j.immuni.2014.12.020 Chen, D. S. (1993). Natural history of chronic hepatitis B virus infection: new light on an old story. J Gastroenterol Hepatol, 8(5), 470-475. Chou, H. H., Chien, W. H., Wu, L. L., Cheng, C. H., Chung, C. H., Horng, J. H., . . . Chen, D. S. (2015). Age-related immune clearance of hepatitis B virus infection requires the establishment of gut microbiota. Proc Natl Acad Sci U S A, 112(7), 2175-2180. doi: 10.1073/pnas.1424775112 Cowie, B. C., Carville, K. S., & Maclachlan, J. H. (2013). Mortality due to viral hepatitis in the Global Burden of Disease Study 2010: new evidence of an urgent global public health priority demanding action. Antivir Ther. doi: 10.3851/IMP2654 Doherty, D. G., & O'Farrelly, C. (2000). Innate and adaptive lymphoid cells in the human liver. Immunological Reviews, 174(1), 5-20. doi: 10.1034/j.1600-0528.2002.017416.x El Kasmi, K. C., Anderson, A. L., Devereaux, M. W., Fillon, S. A., Harris, J. K., Lovell, M. A., . . . Sokol, R. J. (2012). Toll-like receptor 4-dependent Kupffer cell activation and liver injury in a novel mouse model of parenteral nutrition and intestinal injury. Hepatology, 55(5), 1518-1528. doi: 10.1002/hep.25500 Gomez Perdiguero, E., Klapproth, K., Schulz, C., Busch, K., Azzoni, E., Crozet, L., . . . Rodewald, H. R. (2015). Tissue-resident macrophages originate from yolk-sac-derived erythro-myeloid progenitors. Nature, 518(7540), 547-551. doi: 10.1038/nature13989 Gorczynski, R. M. (1992). Immunosuppression induced by hepatic portal venous immunization spares reactivity in IL-4 producing T lymphocytes. Immunol Lett, 33(1), 67-77. Guha, C., Mohan, S., Roy-Chowdhury, N., & Roy-Chowdhury, J. (2004). Cell culture and animal models of viral hepatitis. Part I: hepatitis B. Lab Anim (NY), 33(7), 37-46. doi: 10.1038/laban0704-37 Huang, L.-R., Wu, H.-L., Chen, P.-J., & Chen, D.-S. (2006). An immunocompetent mouse model for the tolerance of human chronic hepatitis B virus infection. Proc Natl Acad Sci U S A, 103(47), 17862-17867. doi: 10.1073/pnas.0608578103 Huang, L. R., Wu, H. L., Chen, P. J., & Chen, D. S. (2006). An immunocompetent mouse model for the tolerance of human chronic hepatitis B virus infection. Proc Natl Acad Sci U S A, 103(47), 17862-17867. doi: 10.1073/pnas.0608578103 Hyams, K. C. (1995). Risks of chronicity following acute hepatitis B virus infection: a review. Clin Infect Dis, 20(4), 992-1000. Ikarashi, M., Nakashima, H., Kinoshita, M., Sato, A., Nakashima, M., Miyazaki, H., . . . Seki, S. (2013). Distinct development and functions of resident and recruited liver Kupffer cells/macrophages. J Leukoc Biol, 94(6), 1325-1336. doi: 10.1189/jlb.0313144 Jiang, W., Wu, N., Wang, X., Chi, Y., Zhang, Y., Qiu, X., . . . Liu, Y. (2015). Dysbiosis gut microbiota associated with inflammation and impaired mucosal immune function in intestine of humans with non-alcoholic fatty liver disease. Sci. Rep., 5. Limmer, A., Ohl, J., Kurts, C., Ljunggren, H. G., Reiss, Y., Groettrup, M., . . . Knolle, P. A. (2000). Efficient presentation of exogenous antigen by liver endothelial cells to CD8+ T cells results in antigen-specific T-cell tolerance. Nat Med, 6(12), 1348-1354. doi: 10.1038/82161 Lyons, S., Sharp, C., LeBreton, M., Djoko, C. F., Kiyang, J. A., Lankester, F., . . . Simmonds, P. (2012). Species association of hepatitis B virus (HBV) in non-human apes; evidence for recombination between gorilla and chimpanzee variants. PLoS One, 7(3), e33430. doi: 10.1371/journal.pone.0033430 Movita, D., Kreefft, K., Biesta, P., van Oudenaren, A., Leenen, P. J., Janssen, H. L., & Boonstra, A. (2012). Kupffer cells express a unique combination of phenotypic and functional characteristics compared with splenic and peritoneal macrophages. J Leukoc Biol, 92(4), 723-733. doi: 10.1189/jlb.1111566 Op den Brouw, M. L., Binda, R. S., Geijtenbeek, T. B., Janssen, H. L., & Woltman, A. M. (2009). The mannose receptor acts as hepatitis B virus surface antigen receptor mediating interaction with intrahepatic dendritic cells. Virology, 393(1), 84-90. doi: 10.1016/j.virol.2009.07.015 Rakoff-Nahoum, S., Paglino, J., Eslami-Varzaneh, F., Edberg, S., & Medzhitov, R. (2004). Recognition of Commensal Microflora by Toll-Like Receptors Is Required for Intestinal Homeostasis. Cell, 118(2), 229-241. doi: http://dx.doi.org/10.1016/j.cell.2004.07.002 Saeed, S., Quintin, J., Kerstens, H. H., Rao, N. A., Aghajanirefah, A., Matarese, F., . . . Stunnenberg, H. G. (2014). Epigenetic programming of monocyte-to-macrophage differentiation and trained innate immunity. Science, 345(6204), 1251086. doi: 10.1126/science.1251086 Suda, T., Gao, X., Stolz, D. B., & Liu, D. (2007). Structural impact of hydrodynamic injection on mouse liver. Gene Ther, 14(2), 129-137. doi: 10.1038/sj.gt.3302865 Vanlandschoot, P., Van Houtte, F., Roobrouck, A., Farhoudi, A., Stelter, F., Peterson, D. L., . . . Leroux-Roels, G. (2002). LPS-binding protein and CD14-dependent attachment of hepatitis B surface antigen to monocytes is determined by the phospholipid moiety of the particles. J Gen Virol, 83(Pt 9), 2279-2289. Visvanathan, K., Skinner, N. A., Thompson, A. J., Riordan, S. M., Sozzi, V., Edwards, R., . . . Locarnini, S. (2007). Regulation of Toll-like receptor-2 expression in chronic hepatitis B by the precore protein. Hepatology, 45(1), 102-110. doi: 10.1002/hep.21482 Wu, J., Meng, Z., Jiang, M., Pei, R., Trippler, M., Broering, R., . . . Schlaak, J. F. (2009). Hepatitis B virus suppresses toll-like receptor-mediated innate immune responses in murine parenchymal and nonparenchymal liver cells. Hepatology, 49(4), 1132-1140. doi: 10.1002/hep.22751 Xu, L., Yin, W., Sun, R., Wei, H., & Tian, Z. (2014). Kupffer cell-derived IL-10 plays a key role in maintaining humoral immune tolerance in hepatitis B virus-persistent mice. Hepatology, 59(2), 443-452. doi: 10.1002/hep.26668 Yoshimoto, S., Loo, T. M., Atarashi, K., Kanda, H., Sato, S., Oyadomari, S., . . . Ohtani, N. (2013). Obesity-induced gut microbial metabolite promotes liver cancer through senescence secretome. Nature, 499(7456), 97-101. doi: 10.1038/nature12347 You, Q., Cheng, L., Kedl, R. M., & Ju, C. (2008). Mechanism of T cell tolerance induction by murine hepatic Kupffer cells. Hepatology, 48(3), 978-990. doi: 10.1002/hep.22395 | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/4527 | - |
dc.description.abstract | 慢性B型肝炎在全球約有3億的慢性帶原者,患者發生感染時的年齡與慢性B肝型肝炎的發展有很大的關聯性,其中有高於90%的新生兒會發展成慢性帶原者,而5歲以後成為帶原者的機率只有5~7%。在過去的研究中,我們發現利用高壓尾靜脈注射使C3H/HeN小鼠模擬B型肝炎病毒感染,可以依據小鼠年齡產生兩種不同的免疫反應。6周歲的小鼠對B型肝炎病毒產生免疫容忍,無法產生有效的免疫反應清除病毒,12周歲小鼠則可在6周內清除病毒,然而12周歲小鼠清除B型肝炎病毒的能力會因為使用抗生素去除腸道菌而降低。觀察小鼠的腸道菌發現,腸道菌的菌相會隨著年齡變化,到12周歲時會達到穩定。許多研究顯示,肝臟的免疫環境會根據外來抗原產生變化,特別是從腸道來的細菌代謝產物會使肝臟成為免疫容忍狀態,降低外來抗原引起的免疫反應,因此我們推論在C3H/HeN小鼠模型中,6周歲小鼠因為當時的腸道菌抗原造成較強的免疫容忍使其無法有效清除B型肝炎病毒,而12周歲小鼠的腸道菌因為組成和6周歲小鼠不同,其產生的抗原組成也有差異,進而造成肝臟的免疫環境足以產生有效的免疫反應來清除病毒。庫佛氏細胞在許多的研究中被認為是造成肝臟免疫容忍的重要免疫細胞,它會過濾外來抗原並產生免疫調控因子來調節免疫反應。我們將6周歲C3H/HeN小鼠的庫佛氏細胞剃除後,發現小鼠有效地清除病毒。接著我們觀察6周歲和12周歲小鼠在接觸B型肝炎後肝臟巨噬細胞族群的變化情形,發現在12周歲小鼠以高壓尾靜脈注射打入B型肝炎病毒質體後第二天,庫佛氏細胞會大量消失,而6周歲小鼠仍保有許多庫佛氏細胞。6周歲C3H/HeJ小鼠為4號類鐸受體突變並且失去功能的品系,我們推測其可能具有較差的肝臟免疫容忍,並證實其的有快速清除HBV的能力。同時我們也觀察到6周歲C3H/HeJ小鼠在高壓尾靜脈注射B型肝炎病毒質體後的18小時以及第2天,庫佛氏細胞也大量消失。由此結果推論,小鼠會藉由剔除庫佛氏細胞來破壞肝臟免疫容忍,進而使肝臟的環境易於產生較強的免疫反應來對抗病毒。最後我們發現腸道菌剃除的12周歲C3H/HeN小鼠,其庫佛氏細胞在高壓尾靜脈注射B型肝炎病毒質體後的第三天大量消失,此時間點在6周和12周歲C3H/HeN小鼠以及6周歲C3H/HeJ小鼠,庫佛氏細胞族群已經回復大多數。因此我們推論高壓尾靜脈注射B型肝炎病毒質體後的第三天,新增的庫佛氏細胞可能和原庫佛氏細胞具有不同的功能,進而幫助清除B型肝炎病毒。
綜合以上結果,我們發現庫佛氏細胞的消失對於小鼠清除B型肝炎病毒可能為必要的過程。B型肝炎病毒透過何種抗原引起庫佛氏細胞的消失,以及庫佛氏細胞是如何接收到B型肝炎病毒的訊息是未來重要的研究方向。 | zh_TW |
dc.description.abstract | Approximately 300 million people are HBV carrier. Mechanisms leading to hepatitis B virus (HBV) persistence is still unclear. One of important factor is the age of HBV infection. More than 90% of newborns or infants who acquire HBV infection become HBV carriers. The risk is reduced to 25~30% in 0~5 years old child and 5~7% after 5 years old. In our previous studies, we established age-related HBV clearance mice model on C3H/HeN mice and indicated that gut microbiota might be an important factor in HBV clearance. It is known that the composition of gut microbiota can change with age and become stable in adult C3H/HeN mice. We suppose that foreign antigens especially derived from gut microbiota can shape liver immune environment into immune tolerance to HBV in young C3H/HeN mice. Whereas, Gut microbiota in adult C3H/HeN provides difference signals from young C3H/HeN mice to influence liver environment and contribute to interrupt liver tolerance at the exposure to HBV. Kupffer cell (KC) is known for its immune regulatory effect in liver tolerance. Therefore we depleted KCs of young and adult C3H/HeN mice before transfection of HBV and found that the depletion of KCs can promote HBV clearance in young C3H/HeN mice. It indicates that the interrupt of liver tolerance may be an important pathway to clear HBV. Next, we investigated the population of liver macrophages in C3H/HeN mice. The population of liver macrophages can be divided into two subset, infiltrating macrophages and KCs, by CD11b and F4/80. We found that KCs dramatically decreased and massive macrophage infiltration after hydrodynamic injection (HDI) of pAAV/HBV1.2 in adult C3H/HeN mice. The change of liver macrophage population was completely restored at day 9 after HDI. However, the reduction of KC in young C3H/HeN mice was not notable than adult C3H/HeN mice and mainly caused by the effect of HDI. These results support our hypothesis that adult C3H/HeN mice have the ability to interrupt liver tolerance by reduction of KC at the exposure to HBV. We have the similar result on C3H/HeJ mice which are TLR4-deficient strand. Young C3H/HeJ mice have the ability to clear HBV that may be due to the weak liver tolerance causing by the loss of TLR4 signaling. As our previous study, the ability of HBV clearance can be partially disrupted by antibiotic treatment in adult C3H/HeN mice. We further investigated the population of liver macrophages in antibiotic-treated adult C3H/HeN mice and found that the population of KCs was significantly reduced compared with young and adult C3H/HeN mice which didn’t received antibiotic treatment. It indicates that the function of KCs at day 3 after the exposure of HBV are opposite to the original KCs before HDI and may promote HBV clearance.
In conclusion, our results suggest that the reduction of KCs is important to promote HBV clearance. It may be due to the interruption of liver tolerance that make liver environment trend to induce stronger innate immune response to against HBV. In the future, to find out what antigens of HBV promote the reduction of KCs is a top priority. That will provide a clue to further investigate the mechanism of age-related immune response to HBV. | en |
dc.description.provenance | Made available in DSpace on 2021-05-14T17:43:01Z (GMT). No. of bitstreams: 1 ntu-104-R02445123-1.pdf: 2744747 bytes, checksum: 86247ff09a8abd9bcd9aa0dc4ed61233 (MD5) Previous issue date: 2015 | en |
dc.description.tableofcontents | 致謝............1
國立臺灣大學碩士學位論文口試委員會審定書............2 中文摘要............3 ABSTRACT............5 CHAPTER 1: INTRODUCTION............8 1.1 Nature history of hepatitis B infection............8 1.2 Animal models of hepatitis B virus infection............8 1.3 Mouse gut microbiota participated in liver immunity against HBV............10 1.4 The role of Kupffer cells in hepatitis B............12 1.4.1 The characteristics of Kupffer cells............12 1.4.2 The role of KCs during HBV infection............13 1.5 Hypothesis and specific aim............15 CHAPTER 2: MATERIAL AND METHODS............17 2.1 Materials............17 2.1.1 Antibodies............17 2.1.2 Reagents............18 2.2 Methods............18 2.2.1 Hydrodynamic HBV transfection mouse model............18 2.2.2 Method for isolation and enrichment of liver NPC............19 2.2.3 Flow cytometry analysis............20 2.2.4 Kupffer cell depletion............21 2.2.5 Sterilization of mouse gut microbiota............21 CHAPTER 3: RESULTS............22 3.1 The depletion of KC promoted HBV clearance of young C3H/HeN mice............22 3.2 Kupffer cell population of adult C3H/HeN mice dramatically decreased after transfection of HBV............23 3.3 The delayed recovery of KCs in antibiotics-treated adult C3H/HeN mice after exposure to HBV............25 3.4 TLR4 contributed to the immune tolerance to HBV in young C3H............26 3.5 Kupffer cell population of young C3H/HeJ mice decreased after transfection of HBV............28 CHAPTER 4: DISCUSSIONS............29 CHAPTER 5: FIGURES............34 Figure 1. Map of HBV construct............34 Figure 2. Flowchart of KC depletion experiment.............35 Figure 3. The efficiency of KCs depletion by using clodronate liposome............37 Figure 4. HBV persistent rate of young C3H/HeN mice was reduced after the depletion of KCs............40 Figure 5. HDI time course of young C3H/HeN mice............44 Figure 6. HDI time course of adult C3H/HeN mice............47 Figure 7. Flowchart of antibiotic experiment............49 Figure 8. KCs of antibiotics-treated adult C3H/HeN mice decreased at day 3 after the exposure to HBV............52 Figure 9. The loss of TLR4 signaling resulted in dramatic reduction of HBV persistent rate.............54 Figure 10. Kupffer cell population of young C3H/HeJ mice decreased after the transfection of HBV............57 Figure 11. Gating strategy of liver macrophages............60 REFERENCE............61 | |
dc.language.iso | en | |
dc.title | 以小鼠模式探討年齡和腸道菌因子以及肝臟巨噬細胞對B型肝炎病毒容忍性之影響 | zh_TW |
dc.title | The role of liver macrophages in age and gut microbiota dependent HBV-persistent C3H/HeN mice | en |
dc.type | Thesis | |
dc.date.schoolyear | 103-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 王弘毅(Hurng-Yi Wang),楊宏志(HUNG-CHIH YANG),葉秀慧(Shiou-Hwei Yeh) | |
dc.subject.keyword | B型肝炎,巨噬細胞,庫佛氏細胞, | zh_TW |
dc.subject.keyword | HBV,macrophage,kupffer cell, | en |
dc.relation.page | 66 | |
dc.rights.note | 同意授權(全球公開) | |
dc.date.accepted | 2015-08-12 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 微生物學研究所 | zh_TW |
顯示於系所單位: | 微生物學科所 |
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