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| 研究生： |
羅丹梓 Tan-Tzu Lo |
|---|---|
| 論文名稱： |
以TLR7配體咪喹莫特為佐劑是否增進老鼠施打子宮頸癌疫苗的免疫效果 The immunological effect of mice after receiving commercial HPV vaccine with TLR7 ligand imiquimod as adjuvant |
| 指導教授： |
鄭力廷
Li-Ting Cheng |
| 學位類別： |
碩士 Master |
| 系所名稱： |
獸醫學院 - 動物疫苗科技研究所 Graduate Institute of Animal Vaccine Technology |
| 畢業學年度： | 109 |
| 語文別： | 英文 |
| 論文頁數： | 47 |
| 中文關鍵詞： | 人類乳突病毒 、疫苗佐劑 、類鐸受體 、咪喹莫特 |
| 外文關鍵詞： | HPV, vaccine adjuvant, toll-like receptor, imiquimod |
| DOI URL： | http://doi.org/10.6346/NPUST202100244 |
| 相關次數： | 點閱：38 下載：0 |
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子宮頸癌是中低收入國家女性死於癌症的主要原因,幾乎所有的子宮頸癌都是由高致癌危險人類乳突病毒所引起. 由於有效率的篩檢方法及人類乳突病毒疫苗, 子宮頸癌可說是最可以預防的癌症之一. 人類乳突病毒疫苗已證實是安全及有效的, 但是需在超過六個月的時間內施打多次, 還有青春期少女的遵醫囑性差, 都是問題. 現在研究的趨勢是要克服這些困難, 並且讓最需要疫苗的低資源國家更容易獲得疫苗. 添加佐劑是可以改進疫苗效果的一種方式. 類鐸受體是一群可以識別病原的受體. 結合類鐸受體可以活化先天及後天的免疫系統. 咪喹莫特是一種免疫調節藥物. 它可以活化第七類鐸受體並誘發一連串的免疫反應. 這些反應使得咪喹莫特有抗病毒及抗腫瘤的能力. 因此, 咪喹莫特很有潛能成為疫苗佐劑. 本研究顯示比起單純只用人類乳突疫苗,肌肉注射人類乳突疫苗及同時表皮使用咪喹莫特可以明顯昇高實驗鼠的IL-1β, IL-8, IL-12 and IgG.
Cervix cancer is the major reason that women die of cancer in low and middle income countries. Oncogenic HPV is the cause of almost all cervical cancer. On account of effective screening methods and HPV vaccines, cervical cancer is one of the most preventable cancers. HPV vaccines are proved to be safe and useful, but there are problems such as the need of low temperature storage, multiple intramuscular doses administered over 6 months and poor adolescent compliance. There are trends in research to overcome these difficulties and to make vaccines more available in low-resource developing countries where they are most needed. Adding adjuvant is a way that can improve outcome of vaccines.
Toll-like receptors (TLRs) are a group of pathogen recognition receptors. The ligation of toll-like receptor (TLR) activates both the innate and adaptive immune systems. Imiquimod, an immunomodulating drug, can activate TLR7 and trigger a series of immune responses. These effects indicate the antiviral and antitumor qualities of imiquimod. Hence, imiquimod demonstrates potential as vaccine adjuvant. This study showed that intramuscular injection with Gardasil in combination with topical imiquimod ointment as adjuvant evoked obvious elevation of IL-1β, IL-8, IL-12 and IgG in mice comparing to Gardasil only.
Abstracts................................................................................................... II
Acknowledgements .................................................................................IV
Contents ...................................................................................................V
1. Introduction ..........................................................................................1
1.1 Background of the study..…………………………………………..1
1.2 Hypothesis………………..…………………………………………3
1.3 Objective of the Study……..………………………………………..3
2. Review of related literatures….………………………………………..4
2.1 Cervical cancer……………..……………………………………….4
2.2 HPV infection and cancer…..……………………………………….5
2.3 The development of HPV vaccines……..…………………………..7
2.4 The limitations of HPV vaccines………..…………………………10
2.5 Adjuvants………………………………..…………………………11
2.6 Imiquimod: a toll-like receptor agonist…..………………………..13
3. Materials and Methods……………………………………………….15
3.1 Materials……………..…………………………………………….15
3.2 Methods………………..…………………………………………..16
3.2.1 Pilot study: intramuscular immunization and sample collection.16
3.2.2 Intradermal immunization and sample collection……………...16
3.2.3 Quantitative analysis of cytokine mRNA using real-time PCR……………………………………………………………………...17
3.2.4 Cellular response analysis……………………………………...19
3.2.5 Antibody titer analysis………………………………………….20
4. Results…………………………………………………..……………21
4.1. Topical imiquimod stimulated some higher cytokine responses….21
4.2. Topical imiquimod caused more production of CD8 T lymphocytes in the early phase of vaccination ……………………………………….22
4.3. Topical imiquimod evoked higher and longer IgG reaction………23
5. Tables and Figures……………………………………………………24
Table 1.The designation of primers of cytokines and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in this study…..24
Figure 1. Vaccination Schedule………………………………………25
Figure 2. Cytokine profiles of the pilot study…...…………………....26
Figure 3. Interleukin-12 expression……..……………………………27
Figure 4. Interferon-γ expression……..………………………………28
Figure 5. Interleukin-6 expression……………………………………28
Figure 6. Interleukin-1β expression…………..………………………29
Figure 7. Interleukin-8 profile………………………..………………30
Figure 8. Cytokine profiles of female BALB/c mice on the 3rd day after 1st intradermal vaccination with or without topical imiquimod…..…….31
Figure 9. Cytokine profiles of female BALB/c mice on the 17th day of experiment, which was the 3rd day after 2nd intradermal vaccination with or without topical imiquimod…………………..……………………….32
Figure 10. Cytokine profiles of female BALB/c mice on the 28th day of experiment, which was the 14th day after 2nd intradermal vaccination with or without topical imiquimod….………………………………………..33
Figure 11. Flow cytometry result of CD4 T lymphocytes……………34
Figure 12. Flow cytometry result of CD8 T lymphocytes……………35
Figure 13. IgG profile analysis by ELISA after intramuscular vaccination………………………………………………………………36
6. Discussion…………………………………………………………….37
7. References……………………………………………………………39
8. Appendix……………………………………………………………..46
9. Personal Information…………………………………………………46
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校外公開2026/07/25