Title Page
Abstract
Contents
Chapter 1. Research backgrounds and purpose 10
1.1. Research backgrounds 10
1.1.1. Cancer immunotherapy 10
1.1.2. Cancer vaccine 12
1.1.3. DC targeting cancer vaccine 13
1.1.4. Cancer vaccine interacting with T cell directly 14
1.2. Purpose of research 15
Chapter 2. Experimental section 19
2.1. Cell culture 19
2.1.1. Cell line culture 19
2.1.2. Primary cell isolation and culture 20
2.2. Preparation of CCM-MPLA-aCD28 21
2.2.1. Isolation of cancer cell membrane 21
2.2.2. Characterization of cancer cell membrane 22
2.2.3. Preparation of CCM-MPLA 22
2.2.4. Preparation of CCM-MPLA-aCD28 23
2.2.5. Characterization of CCM-MPLA-aCD28 23
2.2.6. Analysis of CCM-MPLA-aCD28 composition 24
2.3. In vitro assays 26
2.3.1. In vitro interaction of CCM-MPLA-aCD28s with DCs and T cells 26
2.3.2. In vitro viability test 26
2.3.3. In vitro DC activation 26
2.3.4. In vitro T cell proliferation and activation 27
2.4. In vivo antitumor efficacy of CCM-MPLA-aCD28 29
2.4.1. In vivo DC activation 29
2.4.2. Biodistribution of CCM-MPLA-aCD28 29
2.4.3. In vivo interaction of CCM-MPLA-aCD28 with DCs and T cells 30
2.4.4. In vivo toxicity test 30
2.4.5. In vivo tumor growth 31
2.4.6. TIL analysis 32
2.4.7. Animal study approval 32
2.4.8. Statistical analyses 33
Chapter 3. CCM-MPLA-aCD28 for cancer treatment 34
3.1. Characterization of CCM-MPLA-aCD28 34
3.2. Composition of CCM-MPLA-aCD28 37
3.3. Interaction of CCM-MPLA-aCD28 with DCs 41
3.4. Interaction of CCM-MPLA-aCD28 with T cells 45
3.5. Interaction of CCM-MPLA-aCD28 with DCs and T cells 49
3.6. Biodostribution of CCM-MPLA-aCD28 51
3.7. In vivo toxicity of CCM-MPLA-aCD28 53
3.8. The antitumor efficacy of CCM-MPLA-aCD28 55
3.9. TIL anlysis 58
Chapter 4. Conclusion 60
References 62
Abstract in Korean 67
Figure 1.1. Schematic illustration depicting the stepwise preparation of CCM-MPLA-aCD28. 17
Figure 1.2. Schematic illustration describing the hypothesis of this study. 18
Figure 3.1. Characterization of CCM-MPLA-aCD28. (A) Cryo-transmission electron microscopic images showing the sphere-shaped... 36
Figure 3.2. Composition of CCM-MPLA-aCD28. The percentages of CCMs or CCM-MPLAs that present the tumor antigen peptide (OVA₂₅₇₋₂₆₄)... 39
Figure 3.3. Interaction of CCM-MPLA-aCD28 with DCs. CCM was derived from EG.7-OVA cells. The irrelevant cancer cell membrane... 43
Figure 3.4. Interaction of CCM-MPLA-aCD28 with T cells. CCM was derived from EG7-OVA cells. irCCM was derived from EL4 cells. T cells... 47
Figure 3.5. Interaction of CCM-MPLA-aCD28 with DCs and T cells. CCM was derived from EG7-OVA cells. irCCM was derived from EL4 cells. T... 50
Figure 3.6. Biodistribution of CCM-MPLA-aCD28. (A) Ex vivo fluorescence imaging showing localization of intradermally administered... 52
Figure 3.7. In vivo toxicity of CCM-MPLA-aCD28. (A) Hematoxylin and eosin staining images of the major organs of BALB/c mice treated with... 54
Figure 3.8. The antitumor efficacy of CCM-MPLA-aCD28. CCM was derived from CT26 cells. The irrelevant cancer cell membrane (irCCM) was... 56
Figure 3.9. TIL analysis of CCM-MPLA-aCD28 injected mice. CCM was derived from CT26 cells. The irrelevant cancer cell membrane (irCCM) was... 59