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TIM3에 의한 T 세포 사이토카인 생산 억제 기전과 TIM3 경로억제에 의한 종양 억제 효과
| DC Field | Value | Language |
|---|---|---|
| dc.contributor.advisor | 박선 | - |
| dc.contributor.author | Lee, Mi Jin | - |
| dc.date.accessioned | 2019-10-21T07:17:21Z | - |
| dc.date.available | 2019-10-21T07:17:21Z | - |
| dc.date.issued | 2011-02 | - |
| dc.identifier.other | 11468 | - |
| dc.identifier.uri | https://dspace.ajou.ac.kr/handle/2018.oak/17823 | - |
| dc.description | 학위논문(박사)--아주대학교 일반대학원 :의학과,2011. 2 | - |
| dc.description.tableofcontents | Ⅰ. INTRODUCTION 1 A. TIM3 and its ligand 2 B. The role of TIM3 in Th1 driven immune response 4 C. The cytopalsmic tail of TIM3 and co-inhibitory receptors in T cell activation 7 D. The effect of function of co-inhibitory receptors on antitumor immune response 10 E. Tumor-specific regulatory T cells in antitumor immunity 11 F. Purposes of this research 13 Ⅱ. MATERIALS AND METHODS 14 A. Cells and cell stimulation 14 B. Mice 14 C. RNA isolation and cDNA synthesis 15 D. Construction of plasmids for Tim3-hIg fusion protein expression 15 E. Construction of plasmids for flag tagging TIM3 protein 16 F. Establishment of stable cells expressing TIM3 protein 18 G. Transfection of plasmids 19 H. Preparation of enriched mouse and human CD4+ T cells 19 I. Flow cytometric analysis 20 J. SDS-PAGE and Western blotting 21 K. Real-time reverse-transcription (RT)-PCR 21 L. Luciferase assay 22 M. ELISA 23 Ⅲ. RESULTS 24 A. IL-2 transcription is reduced in T cells expressing TIM3 24 B. Transcriptional activities of AP-1 and NFAT are reduced in TIM3-over expressing Jurkat T cells 30 C. TIM3 expression attenuates the induction of c-Jun in stimulated T cells but not of c-Fos 32 D. TIM3 expression affects the phosphorylation status of NFAT in stimulated T cells 35 E. TIM3 cytoplasmic tail is essential for its inhibitory activity on the expression of IL-2 and IFN-γ 37 F. C-terminal region of TIM3 is essential for suppression of transcriptional activity of AP-1 and NFAT 43 G. Tim3-hIg expressed in tumor cells inhibits in vivo tumor growth 45 H. Tim3-hIg increases the efficacy of prophylactic tumor vaccine 48 I. The frequencies of CD4+CD25+Foxp3+ regulatory T cells in mice injected with Tim3-hIg expressing tumor cells 50 J. Tim3-hIg expression marginally enhances the efficacy of the therapeutic tumor vaccine but not of the chemotherapy 52 Ⅳ. DISCUSSION 54 Ⅴ. CONCLUSION 60 REFERENCES 61 국문요약 79 | - |
| dc.language.iso | eng | - |
| dc.publisher | The Graduate School, Ajou University | - |
| dc.rights | 아주대학교 논문은 저작권에 의해 보호받습니다. | - |
| dc.title | TIM3에 의한 T 세포 사이토카인 생산 억제 기전과 TIM3 경로억제에 의한 종양 억제 효과 | - |
| dc.title.alternative | Mi Jin Lee | - |
| dc.type | Thesis | - |
| dc.contributor.affiliation | 아주대학교 일반대학원 | - |
| dc.contributor.alternativeName | Mi Jin Lee | - |
| dc.contributor.department | 일반대학원 의학과 | - |
| dc.date.awarded | 2011. 2 | - |
| dc.description.degree | Master | - |
| dc.identifier.localId | 569239 | - |
| dc.identifier.url | http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000011468 | - |
| dc.subject.keyword | TIM3 | - |
| dc.subject.keyword | Th1 activation | - |
| dc.subject.keyword | IL-2 | - |
| dc.subject.keyword | Tim3-hIg | - |
| dc.subject.keyword | Tumor vaccine therapy | - |
| dc.description.alternativeAbstract | T cell immunoglobulin- and mucin-domain-containing molecule-3 (TIM3) has been shown to be expressed on Th1 cells and negatively regulates these cell functions. TIM3 has been implicated in the Th1-driven pathogenesis of autoimmune disease and the viral infection. However, molecular mechanisms of TIM3 inhibiting Th1 cells and the efficacy of tumor vaccine expressing Tim3 blocker have not been well studied. In this study, I investigated the molecular mechanisms underlying down-regulation of cytokine production by human TIM3 expression in T cells and the effect of expression of Tim3-hIg, mouse Tim3 pathway blocker, on tumor growth in mice. First, reduced expression of IL-2 and IFN-γ was demonstrated in both primary CD4+ T cells with high level of TIM3 expression and Jurkat T cells-over expressing TIM3. TIM3-over expression reduced the activities of NFAT and AP-1, important transcription factors of IL-2 expression. The expression of c-Jun and dephosphorylation of NFAT were suppressed by TIM3 expression. The 7 a.a from E261 to I267 of TIM3 cytoplasmic tail was critical for down-regulation of cytokine production. The 54 a.a from E261 to P301 was involved in suppression of AP-1 activity and 34 a.a from E268 to P301 in suppression of NFAT activity. On the other hands, I examined the effect of expression of mouse Tim3 pathway blocker, Tim3-hIg. The tumor growth in mice injected with Lewis lung carcinoma (3LL) cells was suppressed and the frequencies of CD4+CD25+Foxp3+ T cells were reduced. Furthermore, injection of Tim3 pathway inhibitor expressing cells significantly enhanced the efficacy of a prophylactic tumor vaccine and marginally enhanced the efficacy of a therapeutic tumor vaccine. However, when given in combination with 5-fluorouracil, a chemotherapeutic agent, therapeutic tumor vaccine capable of Tim3 pathway inhibition had no additional anti-tumor effect. These data demonstrate that TIM3 inhibits Th1-mediated cytokine production through the suppression of AP-1 and NFAT activity and Tim3 pathway inhibitor enhances the efficacy of tumor vaccination and derives anti-tumor immunity | - |
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