新冠肺炎病人週邊血球之單細胞免疫代謝分析

Single-cell immunometabolic profiling of peripheral blood cells in COVID-19 patients

陳仲宇(Chung-Yu Chen)

指導教授 ： 蔡幸真

國立臺灣大學/醫學院/毒理學研究所/碩士(2022年)

https://doi.org/10.6342/NTU202200063

摘要

新冠肺炎至2022年1月14日為止已奪走超過553萬條生命，而且自2019年底已在全世界造成3億2千多萬人感染。科學家們正努力探索新冠病毒的全貌以希望能找出治療方法和研發疫苗來對抗這令人戰慄的敵人。免疫代謝指的是免疫細胞在活化狀態下細胞內代謝路徑的變化。過去有許多文獻發現免疫細胞在面對新冠病毒感染會改變它們的代謝路徑，但目前探討新冠肺炎感染下免疫代謝的縱貫性研究仍很稀少。在此，我們利用質譜流式細胞儀來進行縱貫性研究並探討輕症和重症新冠肺炎病人的免疫代謝。我們使用了26個免疫標記以及17個代謝標的來分析台大醫院和台北市和平醫院新冠肺炎病人的周邊血單核球細胞。我們從7位輕症和2位重症病人取得了35個檢體，並從健康參與者收集了5個檢體做為控制組。我們從這些樣本發現在健康控制組、輕症和重症病人之間，其免疫細胞的組成比例也不盡相同。除此之外，也觀察到在從重症復原的病人中，骨髓衍生細胞的能量生成會從糖解作用轉換成脂肪酸氧化，而且也在重症病人的骨髓衍生細胞發現粒線體損傷。另外，我們在預後不好的病人發現PD-1在CD8+ T細胞有表現量上升的現象，顯示CD8+ T cells上的PD-1表現量可能能作為新冠肺炎預後的指標。而在最後，我們找到一群T細胞MAIT，在重症病人中表現高量的VDAC伴隨著細胞量急劇減少，顯示MAIT上的VDAC表現量可能能用於預測新冠肺炎病人的嚴重度。總結來說，我們從新冠肺炎病人的周邊血細胞發現多個獨特的特徵，提供一個研究並揭開新冠病毒全貌的方向。

關鍵字

新冠肺炎 ； 新冠病毒 ； 免疫代謝 ； 電壓依賴性陰離子選擇性通道1 ； 程式性細胞死亡蛋白-1

並列摘要

COVID-19 has claimed over 5.5 million lives as of January 14th , 2022. The pandemic has been afflicting more than 320 million people worldwide since the end of 2019. Scientists are making efforts to explore the complete face of SARS-CoV-2 in order to develop treatments and vaccines to confront the frightening enemy. Immunometabolism refers to the changes in intracellular metabolic pathways in immune cells during activation. Several studies have discovered that immune cells would shift their metabolic pathways in response to SARS-CoV-2 infection, while the majority of studies examining immunometabolism among SARS-CoV-2 infection lack longitudinal analysis. Here, we demonstrated a longitudinal study on immunometabolism in patients with mild and critical COVID-19 using Cytometry by time of flight (CyTOF). Twenty-six immune cell lineage markers and 17 metabolic markers were utilized to analyze the peripheral blood mononuclear cells (PBMCs) of patients recruited from the National Taiwan University Hospital and the Taipei City Hospital Heping Branch. Thirty-five blood samples were collected from 7 mild and 2 severe COVID-19 patients, while 5 samples were collected from healthy participants as control. We observed that immune cell population compositions were different between healthy controls, mild and severe COVID-19 patients. Moreover, energy source shifting from glycolysis to β-oxidation was found in myeloid cells from a severe patient during convalescence. We also noted impaired mitochondrial function in myeloid cells from patients with severe conditions. Apart from that, we discovered that the patient with a bad prognosis has low levels of PD-1 on CD8+ circulating T cells, which suggested that expression of PD-1 on CD8+ T cells may be a prognostic marker for COVID-19. At last, we found that a subset of T cells, mucosal-associated invariant T (MAIT) cells, expressed high levels of voltage dependent anion channel (VDAC) accompanied by a dramatic decrease in cell percentage in all patients with severe conditions. This finding suggests that levels of VDAC on MAIT cells may be used to predict COVID-19 patients’ outcomes. In summary, we demonstrated several distinct characteristics among PBMCs in COVID-19 patients, which provides a promising direction for future research.

並列關鍵字

COVID-19 ； SARS-CoV-2 ； Immunometabolism ； VDAC ； PD-1

參考文獻

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