| 中文摘要 | 第5-7页 |
| Abstract | 第7-9页 |
| List of Abbreviations | 第14-16页 |
| Introduction | 第16-20页 |
| 1 Background | 第16-17页 |
| 2. Objective of this research | 第17-18页 |
| 3. Significance of this research | 第18-19页 |
| References | 第19-20页 |
| Chapter One:Literature Review | 第20-60页 |
| 1. Introduction | 第20-23页 |
| 2. Ubiquitin | 第23-24页 |
| 3. The 26S proteasome | 第24-26页 |
| 4. The ubiquitin proteasome pathway (UPP) | 第26-29页 |
| 5. Biochemical pathway for ubiquitination and proteasomal degradation | 第29-32页 |
| 6. Constitutive proteasome, immunoproteasome and cancer | 第32-33页 |
| 7. Inhibition of proteasome in cancer therapy | 第33-35页 |
| 8. Ubiquitinated proteins characterization | 第35-38页 |
| 8.1 Use of histidine-tagged ubiquitin | 第36页 |
| 8.2 Use of ubiquitin-associated domain (UBA domain) | 第36-37页 |
| 8.3 Use of ubiquitin-interacting motifs (UIMs) | 第37-38页 |
| 9. Therapeutic cancer vaccines based on ubiquitinated proteins | 第38-49页 |
| 9.1 Autophagy and cross-presentation | 第39-44页 |
| 9.2 DRibble as a potent cancer vaccine | 第44-47页 |
| 9.3 Whole tumor cells vaccine | 第47-48页 |
| 9.4 Cancer vaccine based on tumor cell lysate | 第48-49页 |
| 10. Conclusion | 第49-51页 |
| References | 第51-60页 |
| Chapter Two Ubiquitinated proteins purification by Vx3(A7) protein from tumor cell lysate | 第60-78页 |
| 1. Introduction | 第60-63页 |
| 2. Materials and Methods | 第63-66页 |
| 2.1 Cell culture and reagents | 第63页 |
| 2.2 Tumor cell lysate preparation | 第63页 |
| 2.3 Ubiquitinated proteins purification by TUBEs | 第63-64页 |
| 2.4 Vx3(A7) protein expression and purification | 第64-65页 |
| 2.5 Ubiquitinated proteins purification by Vx3(A7) protein | 第65页 |
| 2.6 Western blotting | 第65-66页 |
| 3. Results | 第66-74页 |
| 3.1 TUBES were not an effective tool to isolate ubiquitinated proteins from tumor cell lysate | 第66-69页 |
| 3.2 Ubiquitinated proteins were effectively isolated from tumor cell lysate by Vx3(A7) protein | 第69-74页 |
| 4. Conclusion | 第74页 |
| 5. Summary | 第74-76页 |
| References | 第76-78页 |
| Chapter Three The ability of ubiquitinated proteins isolated from tumor lysate to induce atumor specific immune response | 第78-97页 |
| 1. Introduction | 第78-80页 |
| 2. Materials and Methods | 第80-83页 |
| 2.1 Mice | 第80页 |
| 2.2 Tumor cell lysate preparation | 第80页 |
| 2.3 Ubiquitinated proteins purification by Vx3(A7) protein | 第80-81页 |
| 2.4 α-Al2O3 nanoparticles preparation | 第81页 |
| 2.5 Detection of immune responses | 第81-82页 |
| 2.6 DRibbles preparation | 第82页 |
| 2.7 Intracellular staining | 第82页 |
| 2.8 Statistical analysis | 第82-83页 |
| 3. Results | 第83-93页 |
| 3.1 Vaccination of Ub-enriched proteins via inguinal lymph nodes induce a stronger immune response than subcutaneous route | 第83-85页 |
| 3.2 Ub-enriched proteins elicit a stronger tumor specific immune response than whole tumor cell lysate and Ub-depleted proteins | 第85-91页 |
| 3.3 Both Ub-enriched proteins and DRibbles induce a strong tumor specific immune response | 第91-93页 |
| 4. Conclusion | 第93页 |
| 5. Summary | 第93-95页 |
| References | 第95-97页 |
| Chapter Four The therapeutic efficacy of ubiquitinated proteins on the established tumormode | 第97-117页 |
| 1. Introduction | 第97-102页 |
| 2. Materials and Methods | 第102-105页 |
| 2.1 Mice | 第102页 |
| 2.2 Tumor cell lysate preparation | 第102页 |
| 2.3 Ubiquitinated proteins purification by Vx3(A7)protein | 第102-103页 |
| 2.4 α-Al2O3 nanoparticles preparation | 第103页 |
| 2.5 Tumor models establishment and immunotherapy experiments | 第103-104页 |
| 2.6 Statistical analysis | 第104-105页 |
| 3. Results | 第105-115页 |
| 3.1 Vaccination of Ub-enriched proteins via inguinal lymph nodes rather than subcutaneous route induces a potent therapeutic efficacy on established tumor model | 第105-110页 |
| 3.2 Ub-enriched proteins vaccine has a significant inhibitory effect on in vivo growth of homologous tumor as well as allogeneic tumor | 第110-115页 |
| 4. Conclusion | 第115页 |
| 5. Summary | 第115-116页 |
| References | 第116-117页 |
| Conclusions and Future Research | 第117-120页 |
| 1. Conclusion | 第117-119页 |
| 1.1 Vx3(A7) protein is an efficient tool to isolate ubiquitinated proteins from tumor cell lysate | 第117-118页 |
| 1.2 Ub-enriched proteins could induce a tumor specific immune response | 第118页 |
| 1.3 Ub-enriched proteins have a potent anti-tumor efficacy in the established tumor models | 第118-119页 |
| 2. Future Research | 第119-120页 |
| About the Author | 第120-121页 |
| Acknowledgement | 第121页 |
