| 摘要 | 第3-4页 |
| abstract | 第4页 |
| ABBREVIATIONS | 第10-11页 |
| Chapter 1 Introduction | 第11-39页 |
| 1.1 Human tumor-associated virus | 第11-20页 |
| 1.1.1 Hepatitis C Virus | 第12-17页 |
| 1.1.2 Human Papillomavirus | 第17-20页 |
| 1.2 Viral Vaccine and Cancer Immunotherapy | 第20-24页 |
| 1.2.1 HCV Vaccine | 第21-22页 |
| 1.2.2 HPV vaccine | 第22-24页 |
| 1.3 Adjuvant for viral vaccine | 第24-31页 |
| 1.3.1 Types of Immunologic Adjuvants | 第25-26页 |
| 1.3.2 Adjuvant Mechanisms of Action | 第26-28页 |
| 1.3.3 Application of Polymer-based nanoparticle adjuvant for viral vaccine | 第28-29页 |
| 1.3.4 Poly(lactic-co-glycolic) acid (PLGA) application to viral vaccine | 第29-31页 |
| 1.4 Dendritic Cells vaccine | 第31-34页 |
| 1.4.1 Therapeutic vaccination via DCs | 第32页 |
| 1.4.2 Ex vivo-generated DC vaccines and Targeting antigen to DCs in vivo | 第32-34页 |
| 1.5 Research goals and outline of dissertation | 第34-39页 |
| 1.5.1 Perspective (I) for HCV Vaccine | 第34-35页 |
| 1.5.2 Perspective (II) for HCV Vaccine | 第35-36页 |
| 1.5.3 Perspective (III) for HCV Vaccine | 第36-37页 |
| 1.5.4 Perspective (IV) for HCV Vaccine | 第37-39页 |
| Chapter 2 Materials and Methods | 第39-58页 |
| 2.1 HCV | 第39-43页 |
| 2.1.1 Materials | 第39页 |
| 2.1.2 Mice | 第39页 |
| 2.1.3 Construction of recombinant expression plasmids | 第39-40页 |
| 2.1.4 Preparation of encapsulated HCV1b-E2 into PLGA microspheres | 第40-41页 |
| 2.1.5 Characterization of HCV1b-E2-PLGA microspheres | 第41页 |
| 2.1.6 HCV1b-E2-PLGA encapsulated efficiency | 第41页 |
| 2.1.7 In vitro release of study | 第41-42页 |
| 2.1.8 Mice Antibody Response and CD8+ T-cell Analysis | 第42页 |
| 2.1.9 IFN-γ ELISA assay | 第42页 |
| 2.1.10 Statistical analysis | 第42-43页 |
| 2.2 HPV (I) | 第43-46页 |
| 2.2.1 Materials | 第43页 |
| 2.2.2 Mice | 第43页 |
| 2.2.3 Antigens | 第43-44页 |
| 2.2.4 Mice immunization | 第44页 |
| 2.2.5 Indirect ELISA | 第44页 |
| 2.2.6 Culture of immature and mature Human Dendritic Cells | 第44-45页 |
| 2.2.7 Analysis of Pre-Cultured Lymphocytes | 第45页 |
| 2.2.8 Pre-mature HuDCs and mature HUDCs co-cultured with Lymphocytes and IFN-γ ELISA assay | 第45-46页 |
| 2.2.9 Cell Viability Assay | 第46页 |
| 2.2.10 Statistical analysis | 第46页 |
| 2.3 HPV(II) | 第46-53页 |
| 2.3.1 Materials | 第46-47页 |
| 2.3.2 Mice | 第47页 |
| 2.3.3 Antigens | 第47-48页 |
| 2.3.4 Preparation of PLGA NPs | 第48页 |
| 2.3.5 Preparation of E7PLGA | 第48页 |
| 2.3.6 Preparation of E7PLGA | 第48页 |
| 2.3.7 Characterization of PLGA NPs and E7PLGA | 第48-49页 |
| 2.3.8 E7 protein quantiation on PLGA NPs | 第49页 |
| 2.3.9 Mice Antibody Response | 第49-50页 |
| 2.3.10 Culture of Human Dendritic Cells | 第50-51页 |
| 2.3.11 Maturation of HuDCs | 第51页 |
| 2.3.12 Cellular uptake of PLGA-C6 NPs in HuDCs | 第51页 |
| 2.3.13 Analysis of T-Lymphocytes before co-culturing with HuDCs | 第51-52页 |
| 2.3.14 Lymphocytes co-cultured with HeLa cells and Cell viability assays | 第52页 |
| 2.3.15 T-lymphocytes co-cultured with HeLa cells and Cell viability assays | 第52-53页 |
| 2.3.16 IFN-γ ELISA assay | 第53页 |
| 2.3.17 Statistical analysis | 第53页 |
| 2.4 HPV(III) | 第53-58页 |
| 2.4.1 Materials | 第53-54页 |
| 2.4.2 Mice | 第54页 |
| 2.4.3 Preparation of encapsulated E7 protein into PLGA microspheres | 第54页 |
| 2.4.4 E7-ENC-PLGA size and morphology | 第54-55页 |
| 2.4.5 Competitive ELISA | 第55页 |
| 2.4.6 E7-ENC-PLGA microspheres encapsulated efficiency | 第55-56页 |
| 2.4.7 In vitro release studies | 第56页 |
| 2.4.8 Mice Antibody Response | 第56页 |
| 2.4.9 CD4+, CD8+ T-cell Analysis | 第56-57页 |
| 2.4.10 Statistical analysis | 第57-58页 |
| Chapter 3 E2 protein into PLGA microspheres could induce mice cytotoxic T-cell response | 第58-68页 |
| 3.1 HCV1b-E2 protein antigen was presented at a molecular mass of 20 kDa | 第58-59页 |
| 3.2 HCV1b-E2-PLGA microspheres morphology and sizes | 第59-60页 |
| 3.3 Encapsulation efficacy | 第60-62页 |
| 3.4 HCV1b-E2 protein released from PLGA microspheres | 第62-63页 |
| 3.5 An expression of mice cytotoxic T-cells | 第63-65页 |
| 3.6 HCV1b-E2-PLGA microspheres could induce IFN-γ production | 第65-66页 |
| 3.7 HCV1b-E2-PLGA could induce humoral immune response | 第66-68页 |
| Chapter 4 Immunotherapeutic effect of dendritic cells with E7/HPV-E7 onto PLGA | 第68-93页 |
| 4.1 E7 protein antigen was presented at a molecular mass of 20 kDa | 第68-69页 |
| 4.2 TEM demonstrated PLGA NPs morphology and E7 protein was actually bound to PLGA NPs | 第69-70页 |
| 4.3 E7 protein and E7PLGA could induce humoral immune response | 第70-71页 |
| 4.4 PLGA NPs were uptaken by PLGA NPS | 第71-72页 |
| 4.5 Maturation of HuDCs | 第72-73页 |
| 4.6 T-lymphocytes number were analyzed before co-culturing with pre-mHuDCs and mHuDCs | 第73-74页 |
| 4.7 T-lymphocytes number were analyzed before co-culturing with HuDCs pulsed with E7 protein and E7PLGA | 第74-76页 |
| 4.8 E7 protein could induce IFN-γ production in both pre-mHuDCs and mHuDCs pulsed with E7 protein | 第76-77页 |
| 4.9 E7PLGA could induce IFN-γ production | 第77-78页 |
| 4.10 CTL activity to kill the cervical cancer cells is induced by pre-mHuDCs and mHuDCs pulsed with E7 protein | 第78-81页 |
| 4.11 CTL activity to kill the cervical cancer cells is induced by E7PLGA | 第81-85页 |
| 4.12 TEM demonstrated E7-ENC-PLGA morphology and size | 第85页 |
| 4.13 E7PLGA and E7-ENC-PLGA protein quantification | 第85-86页 |
| 4.14 Encapsulation efficientcy | 第86-88页 |
| 4.15 E7 protein released from PLGA microspheres | 第88-89页 |
| 4.16 E7PLGA could induce higher humoral immune response | 第89-90页 |
| 4.17 An expansion of helper T-cells was triggered by E7PLGA | 第90-93页 |
| Chapter 5 Conclusions and Prospect | 第93-99页 |
| 5.1 Hepatitis C virus E2 protein encapsulation into PLGA microspheres could induce mice cytotoxic T-cell response | 第93-95页 |
| 5.2 Immunotherapeutic Effect of Pre-mature and Mature Human Dendritic Cells Pulsed with HPV16-E7 Protein | 第95-96页 |
| 5.3 Immunotherapeutic Effect of Human Dendritic Cells Pulsed with HPV16-E7 Protein Adsorbed onto PLGA | 第96-97页 |
| 5.4 Effect of PLGA microspheres with protein adsoption and encapsulation to T-lymphocytes | 第97-99页 |
| References | 第99-110页 |
| Acknowledgements | 第110-112页 |
| Curriculum Vitae | 第112页 |
