臺灣博碩士論文加值系統

SARS-CoV-2是嚴重特殊傳染性肺炎（Coronavirus disease 2019, COVID-19）的病原體，在全球造成嚴重的疫情，其在個體之間所造成的疾病不盡相同。雖然大多數COVID-19患者表現為無症狀或輕至中度的疾病，但有些個案會出現「長新冠後遺症」，其定義為患者初次感染SARS-CoV-2後3個月內繼續或出現新症狀，並且這些症狀持續至少2個月且沒有其他解釋，確切機制目前不清楚。最近的研究表明，在疾病後期嚴重COVID-19病例中的過度訓練型免疫會導致免疫失調，除此之外在部分患者，即便症狀很輕微也會表現出過度發炎，尤其是伴隨IL-6水平升高。長期以來，人們認為免疫記憶是適應性免疫反應的獨特標誌，然而有越來越多的文獻表明先天免疫細胞在某些感染或疫苗接種後也可以表現出適應性特徵，對隨後獨立於適應性免疫的挑戰產生強化且持久的免疫反應。就目前的研究表明，先天免疫記憶在COVID-19中的作用有兩種說法：一是有助於防止再次感染或減輕繼發病毒感染的嚴重性，二是過度強烈地誘導訓練有素的免疫力可能會導致COVID-19患者的長期炎症和產生其他併發症。因此我們假設與長新冠後遺症相關的免疫失調和多器官功能障礙導致持續性炎症是緣於先天免疫記憶的調節，因此本研究目的主要為了解SARS-CoV-2如何誘導先天免疫記憶，以及這種記憶在長新冠後遺症中起保護作用或加劇感染後的發炎反應。
我們的實驗結果初步顯示，單獨使用SARS-CoV-2 spike protein在初次刺激後對於隨後的刺激無法產生先天免疫記憶，而使用polyI:C或LPS作為先天免疫誘導劑進行刺激對隨後SARS-CoV-2 spike protein的刺激可以表現出記憶反應，這種記憶主要表現為增加產生IL-6的能力。此外，我們還發現當SARS-CoV-2 spike protein與其他刺激物共同刺激時，可以作為先天免疫記憶的誘導劑，然而可能潛在地抑制其他刺激物誘導先天免疫記憶的能力。
我們研究的主要發現SARS-CoV-2的結構蛋白－spike protein，可以在人類細胞中誘導訓練型免疫。儘管我們的實驗結果只能提供基礎的想法，未來還需要進行更多的實驗來驗證和鞏固我們的結論。

SARS-CoV-2, the virus causing the global Coronavirus disease 2019 (COVID-19) pandemic, affects individuals differently. While most COVID-19 cases result in asymptomatic or mild-to-moderate illness, some experience "Long COVID." This condition occurs when patients continue or develop new symptoms for at least two months after their initial infection, and its exact mechanisms remain unclear. Recent research has shown that an excessive trained immunity in severe COVID-19 cases leads to immune dysregulation, particularly in later stages. Some patients, even with mild cases, exhibit excessive inflammation with elevated cytokine levels, notably interleukin-6 (IL-6). Traditionally, immune memory has been considered an acquired immune response feature. However, emerging evidence suggests that the innate immune system can adapt after prior encounters with pathogens. In the context of COVID-19, two perspectives have emerged: innate immunity might contribute to protection against reinfection or worsen long-term inflammatory complications in "Long COVID." Patients with "Long COVID" often show transcriptional dysregulation and immune dysfunction in their innate immune cells. This suggests that immune dysregulation and multi-organ dysfunction during acute COVID-19 may be linked to trained immunity.
We would like to explore the role of trained immunity during SARS-CoV-2 infection. We aim to understand how SARS-CoV-2 triggers trained immunity and whether this memory influences "Long COVID," protective responses, or post-infection outcomes.
Our experimental results preliminarily indicate that utilizing SARS-CoV-2 spike protein following the initial stimulation failed to induce trained immunity. Cells trained with LPS and polyI:C exhibit a memory response upon subsequent stimulation with SARS-CoV-2 spike protein. This memory was almost exclusively represented by an increased capacity to produce IL-6. Furthermore, we found that spike protein can also serve as an inducer of innate immune memory when combined with other stimuli. However, it may inhibit other stimuli's ability to induce trained immunity.
The major finding of our study is that one of the SARS-CoV-2 proteins, spike protein, can elicit trained immunity in human cells. Although our initial experimental findings provide a foundation, additional experiments are imperative to validate and solidify our conclusion.

總目錄 I
正文目錄 II
表目錄 V
圖目錄 VI
中文摘要 VIII
Abstract X
第一章 緒論 1
第一節 嚴重急性呼吸道症候群冠狀病毒2型 1
第二節 先天性免疫 8
第三節 研究動機與目的 13
第二章 實驗材料及方法 15
第一節 建立體外先天免疫記憶模型 15
第二節 誘導先天免疫記憶後所產生之抗病毒和發炎反應分析 21
第三節 先天免疫記憶中第一型干擾素的分析 22
第三章 結果 26
第一節 建立體外先天免疫記憶模型 26
第二節 利用人類免疫和非免疫細胞評估SARS-CoV-2 spike protein誘導先天免疫記憶的能力 28
第三節 探討第一型干擾素在先天免疫記憶中的作用 35
第四章 討論 37
第一節 利用不同的檢測方法來分析先天免疫記憶的產生 37
第二節 不同SARS-CoV-2變異株spike protein之間的差異 38
第三節 SARS-CoV-2或其成分在先天免疫記憶中的角色 39
第四節 SARS-CoV-2 spike protein、polyI:C和LPS對NFκB p65
及IRF3的影響 40
第五章 結論 43
第六章 參考資料 45

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