Eunju Park. (2025). T cell-derived small extracellular vesicles targeting CD24 enhance anti-tumor effect through dual activation of innate and adaptive immunity. doi: 10.22677/THESIS.200000829162
Type
Thesis
Description
Immune checkpoint blockade, PD-L1, CD24, tumor microenvironment, small extracellular vesicle, cancer immunotherapy
Table Of Contents
I. Introduction 1 II. Materials and methods 5 2.1 Phage display panning 5 2.2 Production and purification of antibodies 5 2.3 Enzyme-linked immunosorbent assay (ELISA) 6 2.4 Cell culture 6 2.5 Macrophage differentiation 7 2.6 RNA isolation and quantitative real time-polymerase chain reaction (qRT-PCR) 7 2.7 Macrophage phagocytosis assay 7 2.8 Generation and production of anti-CD24 antibody-engineered T cell-derived sEVs 8 2.9 Staining of sEVs 9 2.10 Characterization of sEVs 9 2.11 Western blotting 10 2.12 Immunocytochemistry (ICC) 11 2.13 In vitro flow cytometry analysis 11 2.14 In vivo flow cytometry analysis 12 2.15 In vivo sEVs targeting assay 13 2.16 In vivo anti-tumor effects in the LLC subcutaneous model 13 2.17 Macrophage and CD8+ T cell depletion 13 2.18 In vivo anti-tumor effects in the LLC-lung metastasis model 14 2.19 Statistical analysis 14 III. Results 15 3.1 Development and characterization of monoclonal antibodies targeting CD24 15 3.2 Production and characterization of αCD24-expressing T cell-derived sEVs 22 3.3 Specific binding and internalization of αCD24-sEVs in cancer cells 25 3.4 Enhancement of macrophage-mediated phagocytosis by αCD24-sEVs 25 3.5 αCD24-sEVs reduce PD-L1 levels in cancer cells 29 3.6 αCD24-sEVs specifically delivered to the tumor in the LLC subcutaneous model 29 3.7 Anti-tumor activity of αCD24-sEVs in the LLC subcutaneous model 32 3.8 αCD24-sEVs enhance phagocytosis and anti-tumor immune responses within the TME 34 3.9 αCD24-sEVs reduce PD-L1 expression within the tumor tissue 34 3.10 The anti-tumor immune response of αCD24-sEVs is dependent on macrophages and CD8+ T cells 37 3.11 Therapeutic efficacy of αCD24-sEVs in an LLC-lung metastasis model 37 IV. Discussion 40 V. References 42 요 약 문 44
