The role of Ets1 in CD8 T cells in association with anti-tumor activity
- 주제(키워드) 도움말 Ets1 , CD8+ T cells , cytokine production , tumor immune surveillance
- 발행기관 강릉원주대학교 일반대학원
- 지도교수 도움말 Dr. Choong-Gu Lee and Prof. Dae-Hee Lee
- 발행년도 2024
- 학위수여년월 2024. 2
- 학위명 석사
- 학과 및 전공 도움말 일반대학원 KIST강릉분원학.연협동과정
- 세부분야 해당없음
- 실제URI http://www.dcollection.net/handler/kangnung/000000011671
- UCI I804:42001-000000011671
- 본문언어 영어
초록/요약 도움말
E26 oncogene homolog 1 (Ets1) serves as a crucial transcription factor in directing the development and function of T cells, particularly the CD8 subset. Motivated by the promising potential inherent in exploring the interplay between Ets1 and CD8 in modulating immune responses for effective tumor control, our study endeavors to delve deeper into this relationship. The dLck-cre(+) Ets1fl/fl mice, which contain the distal lck promoter that drives-Cre expression initiated at the late stage of T cell development in the thymus, were generated. Therefore, it facilitates studying the function of gene of interest in mature T cells. Firstly, splenocytes from littermate control mice (LMC, dLck-cre(-) Ets1fl/fl) and Ets1 conditional knockout mice (cKO, dLck-cre(+) Ets1fl/fl) were isolated and stimulated with Phorbol 12-myristate 13-acetate (PMA)/ionomycin, plate-coated αCD3/αCD28, or Dynabeads™ Mouse T-Activator CD3/CD28 in the presence of transport protein inhibitors. Flow cytometry was performed using a staining panel of surface markers and intracellular cytokines (IFN-γ, TNF-α, IL-10) to investigate cytokine production. The data showed that the frequency of CD8+ T cells was significantly impaired in Ets1 cKO mice compared to LMC. Interestingly, the level of IFN-γ expression in CD8+ T cells was remarkably higher in Ets1-deficient mice than in control groups. In addition, the same tendency was demonstrated when investigating cytokine production in isolated pan-T cells, including CD4+ and CD8+ T cells. For the in vivo experiment, the naïve CD8+ T cells isolated from LMC and Ets1 cKO mice were injected into CD8α KO mice, eliminating CD8 T cell functions. Besides, the pancreatic cancer cell line (Panc02) was injected subcutaneously into these mice. This study demonstrated a significant inhibition of tumor growth in mice receiving naïve CD8+ T cells isolated from cKO mice. Overall, these findings indicate that Ets1 modulated the production of IFN-γ in CD8+ T cells in peripheral systems and enhanced the CD8-driven anti-tumor immune response in vivo.
more목차 도움말
Abstract i
국문 초록 iii
Abbreviations v
Contents vii
List of Figures ix
1. INTRODUCTION 1
2. METHOD AND MATERIALS 14
2.1. Mice 14
2.2. Tissue and cell preparation 14
2.3. Naïve pan-T cell isolation 15
2.4. CD8 T cell isolation and checking purification 16
2.5. CD4 T cell isolation and checking purification 16
2.6. Tumor cell lines and culture 17
2.7. In vivo tumor graft 17
2.8. Tumor measurement 18
2.9. Intracellular cytokine productions 18
2.10. Flow cytometry 19
2.11. ELISA 20
2.12. Apoptosis assay 20
2.13. Statistical analysis 21
3. RESULTS 22
3.1. Expression pattern of TCR-β, CD4, and CD8 T cells in splenocytes from Ets1ΔdLck mice 22
3.2. Apoptosis of CD4 and CD8 T cells in Ets1ΔdLck mice 29
3.3. Cytokine production in splenocytes from Ets1ΔdLck mice 35
3.4. Expression pattern of TCR-β, CD4, and CD8 T cells in pan-T cells from Ets1ΔdLck mice 51
3.5. Cytokine production in pan-T cells from Ets1ΔdLck mice 54
3.6. Tumor growth inhibition in Ets1ΔdLck mice in vivo 62
4. DISCUSSION 70
5. SUMMARY 79
References 80
Acknowledgment 88
