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Activation of CD8+ Cytotoxic T Lymphocytes against Tumor Cells using a TLRL-MUC1-Tn Cancer Vaccine
Table of Contents
Abstract
Acknowledgements
Table of Contents
List of Tables
List of Figures
List of Abbreviations
1. Introduction
1.1.Tumor Antigens
1.2.Immune Mechanism of Tumor Eradication
1.3.Cancer Immunotherapy
1.3.1. Passive Immunization
1.3.2. Active Immunization
1.4.Antibody-Mediated APC Targeting Liposomal Vaccine
1.4.1. MUC Glycopeptide Containing a GalNAc-O-Thr (Tn) TACA (Glycopeptide Tumor Antigen)
1.4.2. Toll-like Receptor Ligand (TLRL)
1.4.3. DPPC (Liposome) 7
1.4.4. L-rhamnose(Xenoantigen)
1.5.Molecular Strategy for Activation of Cytotoxic T Lymphocytes (CTLs)
1.5.1. Glycopeptide-Binding in Complex with MHC class I of MUC1-8-5 10 GalNAc
1.5.2. Aberrantly Glycosylated MUC1 Tripartite Vaccine
1.5.3. Vaccine Design 15
2. Materials and Methods
2.1.Vaccine
2.2.Mouse Model
2.3.DCs Preparation
2.3.1. Isolation of Bones from Mice
2.3.2. Extraction of Mononuclear Cells from Four Bones
2.3.3. Bone Marrow Cell Culture with GM-CSF
2.4. Spleen Cell Suspension
2.4.1. Removal of Spleen
2.4.2. Spleen Cell Suspension
2.5.Tumor Cells
2.5.1. GFP-P815
2.5.2. EL-4
2.6.Isolation of CD4+ and CD8+
2.7. Intracellular Cytokine Staining Assay
2.7.1. Intracellular Cytokine, Interferon Gamma (IFN-γ) Staining
2.7.2. Flow Cytometry
2.8. Killing Assay
2.8.1. Killing Assay Using Flow Cytometry
2.8.2. JAM Assay Using 3H-Thymidine
2.9. ELISA
2.9.1. Anti-Rhamnose Antibody ELISA
2.9.2. Interferon Gamma (IFN-γ) ELISA
2.10. [3H]-Thymidine Incorporation Proliferation Assay
3. Results and Discussion
3.1. Vaccine Test
3.2. New Pam3Cys MUC1-Tn Vaccine Tests
3.2.1. Anti-Rha Antibody Production
3.2.2. Anti-MUC1 Antibody Production
3.2.3. CD8+ Proliferation Assay
3.2.4. CD8+ Cytokine Production
3.2.5. Small Peptide (8 Amino Acids Residue of New MUC1 Tn) Test
3.2.6. JAM Assay
3.3. Discussion
References Bài viết liên quan
