臺灣博碩士論文加值系統

腸病毒七十一型可在嬰幼兒引發重症，症狀包括致命的腦幹腦炎以及肺水腫，然而目前對其致病機轉了解仍十分有限。臨床研究腸病毒七十一型感染引起重症的案例，發現病人在血清和腦脊髓液中含大量的介白素-6，且濃度顯著高出在輕症病人血清和腦脊髓液所測得的量。重症病人在接受靜脈注射免疫球蛋白治療後，血清中介白素-6濃度顯著下降，這些證據皆指出介白素-6在腸病毒七十一型感染中扮演一定的角色。然而目前沒有針對介白素-6在腸病毒感染中所扮演的角色的探討，所以我們利用小鼠感染腸病毒的模式來研究此問題。實驗結果發現，野生型小鼠感染後，血清和腦中皆可以測得介白素-6的增加，且增加的趨勢與小鼠疾病嚴重程度及死亡率成正比。在小鼠感染3 × 105溶菌斑形成單位的病毒後，介白素-6缺陷小鼠的後肢嚴重癱瘓，而小鼠存活率只有18%；在野生型小鼠感染後，後肢癱瘓的狀況顯著比缺陷鼠輕微，小鼠的存活率為63%，明顯高於介白素-6缺陷小鼠。另外，介白素-6缺陷小鼠在感染後第五天的脊髓，以及感染後第七天的肝、脾、腎、腦 (不包含腦幹)，以及腦幹中所含的平均病毒量皆顯著高於野生型小鼠。感染後，野生型小鼠比介白素-6缺陷小鼠有較高濃度的專一性中和抗體，以及較多數量的CD19+及CD19+CD138+ B淋巴球，上述實驗結果顯示介白素-6可藉由增加B淋巴球的增生和活化，及抗體的產生來幫助小鼠清除腸病毒七十一型的複製。除了B淋巴球之外，介白素-6缺陷小鼠的CD4+以及CD8+ T淋巴球的增生以及活化也比野生型小鼠低，顯示介白素-6也可幫助T淋巴球的增生和活化來保護小鼠。在中樞神經系統中，淋巴球浸潤的數量並不受小鼠具不具有介白素-6影響，而介白素-6缺陷小鼠比起野生型小鼠，腦幹中神經受損的狀況較為嚴重。因此，綜合以上實驗，我們的實驗結果顯示介白素-6在腸病毒七十一型的感染中，可藉由幫助CD4+、CD8+ T淋巴球以及B淋巴球的活化、專一性抗體的產生、以及減少腦幹中神經的損傷來達到保護小鼠的效果。

Enterovirus 71 (EV71) causes fatal brain stem encephalitis and cardiopulmonary failure in infants and young children, but its pathogenesis remains unclear. Previous studies showed that in the serum and cerebrospinal fluid specimens of EV71-infected patients with fatal symptoms, levels of interleukin-6 (IL-6) were significantly elevated. Additionally, in the patients with fatal symptoms and treated with intravenous immunoglobulin, serum IL-6 level was significantly reduced. These studies indicated that IL-6 may play a role in EV71 infection. There is no study on the role of IL-6 in EV71 infection; therefore, we used a murine model to address this issue. Our results show that IL-6 was induced in the sera and brains and positively correlated with the mortality rate and disease severity of infected mice. We also compared the mortality rates of wild-type and IL-6-deficient mice. After infection, IL-6-deficient mice displayed severe paralysis in their hind limbs with a survival rate of 18% while wild-type mice displayed significantly less severe hind limb paralysis with a survival rate of 63%. The mean viral loads in the spinal cords at day 5 post-infection and in livers, spleens, kidneys, brains without brain stem regions, and brain stems at day 7 post-infection of infected IL-6-deficient mice were significantly higher than those of wild-type mice. After infection, the titer of virus-specific antibody with neutralizing activity and the numbers of CD19+ B cells and activated (CD19+CD138+) B cells were lower in IL-6-deficient mice than those in wild-type mice, indicating that IL-6 protected mice from infection by enhancing B cell proliferation and activation and subsequent antibody production to reduce viral loads in mouse organs. Besides B cells, the proliferation and activation of CD4+ and CD8+ T cells were reduced in the spleens of IL-6-deficient mice compared with those of wild-type mice, showing that IL-6 also enhanced T cell responses to protect mice. In the central nervous system, the infiltrations of lymphocytes were not much different between mice with or without IL-6, whereas more severe neuron damage was observed in the brain stems of IL-6-deficient mice. Thus, our results show that IL-6 protected mice from EV71 infections by increasing CD4+ T, CD8+ T, and CD19+ B cell proliferation and activation, inducing virus-specific antibody production, and reducing neuron damage in the central nervous system.

Chinese abstract…………………………………………………………3
English abstract…………………………………………………………5
Acknowledgements……………………………………………………...7
Introduction…………………………………………………………….11
Materials and methods………………………………………………...16
Results…………………………………………………………………..20
Discussion………………………………………………………………28
References………………………………………………………………31
Figures………………………………………………………………….37

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