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研究生： 蘇婉淳
Su, Wan-Chun
論文名稱： 研究治療丙型肝炎時病人紅血球 Lewis 抗原之表現
Expression of RBC Lewis antigens in the treatment of HCV
指導教授： 張權發
Chang, Chuan-Fa
學位類別： 碩士
Master
系所名稱： 醫學院 - 醫學檢驗生物技術學系
Department of Medical Laboratory Science and Biotechnology
論文出版年： 2023
畢業學年度： 111
語文別： 英文
論文頁數： 198
中文關鍵詞： 丙型肝炎 、直接抗病毒藥物 、醣蛋白 、Lewis抗原
外文關鍵詞： HCV, DAA, Glycoprotein, Lewis antigen
相關次數： 點閱：57  下載：1 
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    • 肝病是臺灣的國病，如何降低丙型肝炎感染的流行率與死亡率是當前的挑戰。目前現有檢測項目包括：Anti-HCV、HCV RNA或Core antigen test。在丙型肝炎治療初期，干擾素的治癒率約為50%，目前有直接抗病毒藥物（DAAs），其治癒率可達99%，不但改善了傳統治療方法的缺陷，還明顯提高了療效。然而，抗病毒藥物可能會提高肝硬化患者演變為肝細胞癌（HCC）的機率，另外並導致HCC復發。因此，長期觀察丙型肝炎患者對於新治療效果的反應性是必要的，我們需要新的生物標誌物用於丙型肝炎的治療與追蹤。從我們收集病例中，我們確實發現一些患者在過去服用DAA藥物後有復發的問題。醣蛋白已成為許多疾病的有價值的生物標誌物，血型抗原也是一種醣蛋白，像：ABO、H和Lewis系統抗原是醣脂上的醣蛋白和寡醣。由於大部分血液醣蛋白由肝臟所產生的，而一些血型抗原如Lewis抗原與病毒感染高度相關，因此我們希望了解Lewis抗原（如Lea和Leb）到底是否可以作為丙型肝炎治療時的生物標誌物。我們透過Lewis抗原試驗、流式細胞儀和ELISA檢測紅血球與血漿中Lea和Leb的表達。並分析了44例經抗病毒藥物治療丙型肝炎患者，初步發現不同患者治療階段紅血球上或血漿中Lea和Leb的表現確實有差異。本研究的發現不僅有助於理解Lewis抗原與丙型肝炎治療過程之間的關係，也有助於揭示其病理生理意義和分子機制，並未來實現個體化用藥打好基礎。

    Liver disease is widespread in Taiwan; therefore, it is imperative to reduce its prevalence and mortality. Currently available testing methods include Anti-HCV, HCV RNA, or core antigen tests. In the early stage of hepatitis C treatment, the cure rate of interferon is about 50%, while those for direct antiviral drugs (DAAs) can reach 99%. This not only ameliorates the deficits of traditional treatment methods, but also significantly improves their curative effect. However, DAAs may increase the incidence: of hepatocellular carcinoma (HCC) in patients with cirrhosis, causing their relapse. Therefore, long-term observations of hepatitis C patients regarding their response to novel treatments is critical to fully understanding drug effectiveness, requiring innovative biomarkers for continuous follow-up and feedback. In the patient samples we collected, some have indeed registered relapses after DAA treatment. Glycoproteins have become valuable biomarkers for many diseases. Blood group antigens are also a type of glycoprotein; for example, ABO, H and Lewis System antigens are glycoproteins and oligosaccharides on glycolipids. Since most blood glycoproteins are produced by the liver, and some blood group antigens such as Lewis antigens are highly related to viral infections, we want to know whether Lewis antigens (such as Lea and Leb) can be used as biomarkers for hepatitis C treatment. We detected the expression of Lea and Leb in red blood cells and plasma by Lewis antigen tests, flow cytometry and ELISA. After analyzing 44 cases of hepatitis C patients who were treated with antiviral drugs, we initially found that the expression of Lea and Leb on red blood cells or in plasma was indeed different during various treatment stages. The findings of this study not only assist us in understanding the relationship between Lewis antigens and the treatment of hepatitis C, but also help to reveal its pathophysiological significance and molecular mechanism, laying a solid foundation for the realization of individualized medicine.

    摘要 i ABSTRACT ii ACKNOWLEDGEMENTS iii TABLE OF CONTENTS iv ABBREVIATION vii INTRODUCTION 1 Overview Of HCV 1 HCV Epidemiology 2 HCV Genotypes 2 HCV Testing 3 HCV Treatment 4 Effect Of Direct-Acting Antiviral Drugs (DAAs) On HCV Treatment 4 Limitations On HCV Treatment and Testing 5 HCV And Personalized Medicine 6 Glycoproteins Biomarkers 6 A Summary Of ABO, H, And Lewis Blood Group Systems 8 Lewis Blood Group System 9 Correlation Between ABO, H, And Lewis Blood Group Systems and Disease Treatment 9 OBJECTIVE 11 Study objective and Specific Aim 11 Experimental Flow Chart 12 MATERIALS AND METHODS 13 Sample collection 13 Clinically data 13 ABO Typing test 14 Lewis antigen A&B assay（Antigen agglutination test）15 Lewis antigen A&B assay（FLOW test）16 Lewis antigen A&B assay（ELISA test）17 Statistical 17 RESULTS 18 Summary of patient demographics and serological characteristics 18 HCV and blood type correlation 18 HCV viral load changes during DAA treatment 19 Related laboratory data changes during DAA treatment 20 HCV patient Lewis antigen changes during DAA treatment 20 DISCUSSION 22 REFERENCE 24 TABLE 34 TABLE.1. Summary of patient demographics and serological characteristics 35 TABLE.2. HCV patient Lewis antigen changes during DAA treatment 37 FIGURE 38 FIG.1. HCV and blood type correlation 38 FIG.2. HCV viral load changes during DAA treatment 39 FIG.3. GOT performance changes during DAA treatment 40 FIG.4. GPT performance changes during DAA treatment 41 FIG.5. T-Bili performance changes during DAA treatment 42 FIG.6. D-Bili performance changes during DAA treatment 43 FIG.7. AFP performance changes during DAA treatment 44 FIG.8. r-GT performance changes during DAA treatment 45 FIG.9. HbA1c performance changes during DAA treatment 46 FIG.10. RBC performance changes during DAA treatment 47 FIG.11. Hb performance changes during DAA treatment 48 FIG.12. PLT performance changes during DAA treatment 49 FIG.13. PT performance changes during DAA treatment 50 FIG.14. Lewis A Ag qualitative performance changes during DAA treatment 51 FIG.15. Lewis B Ag qualitative performance changes during DAA treatment 52 FIG.16. Lewis A Ag FLOW performance changes during DAA treatment 53 FIG.17. Lewis B Ag FLOW performance changes during DAA treatment 54 FIG.18. Lewis A Ag ELISA performance changes during DAA treatment 55 FIG.19. Lewis B Ag ELISA performance changes during DAA treatment 56 SUPPLEMENTAL FIGURE 57 Supplemental Figure 1. IRB 57 Supplemental Figure 2. Antigen agglutination test procedure 58 Supplemental Figure 3. Antigen agglutination test (Titer definition) 58 Supplemental Figure 4. Flow cytometer procedure 59 Supplemental Figure 5. Flow cytometer control definition 60 Supplemental Figure 6. Gating Strategy RBC Populations 62 Supplemental Figure 7. Flow cytometer data changes of 44 patients during DAA treatment 63 Supplemental Figure 8. Lewis A ELISA（CA19-9）195 Supplemental Figure 9. Lewis B ELISA（MUCA5）197

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