{"id":{"repo_id":"ajou","oai_identifier":"oai:repository.ajou.ac.kr:201003/8590"},"canonical_url":"https://search.dev.ndltd.org/etd/ajou/oai:repository.ajou.ac.kr:201003/8590","repository":{"repo_id":"ajou","name":"Ajou University","base_url":"http://repository.ajou.ac.kr/oai/request"},"display":{"title":"Upstream conserved sequence element that regulates TIM-3 induction in CD4+ T cell","abstract":"The T cell immunoglobulin domain and mucin domain-3 (TIM-3) was discovered in Th1 and has been known as a T cell regulatory molecule. TIM-3 expression is induced by T cell activation through mechanism not yet well-investigated. In the study, I wanted to find DNA region involved in regulation of TIM-3 transcription in human CD4+ T cells between -70 kbp to -1 bp from the TIM-3 transcription start site. Given that regulatory DNA regions show nucleotide sequence homology between species, four conserved non-coding sequences (CNS -29, CNS -53, CNS -56 and CNS -63) were analyzed by comparison human TIM-3 upstream DNA sequences with mouse and cow counterparts. Only at CNS -63, H3K4 dimethylation that is associated with transcriptional activation was observed in Jurkat T cells and CD4+ T cells isolated from human blood. Further, CNS -63 increased luciferase reporter gene expression downstream of TIM-3 minimal promoter. Enhancer activity of CNS -63 was abrogated by mutation of putative NF-κB binding sites between -63423 and -63433. These results indicate that TIM-3 CNS -63 may regulate TIM-3 transcription in human CD4+ T cells.","abstract_html":"The T cell immunoglobulin domain and mucin domain-3 (TIM-3) was discovered in Th1 and has been known as a T cell regulatory molecule. TIM-3 expression is induced by T cell activation through mechanism not yet well-investigated. In the study, I wanted to find DNA region involved in regulation of TIM-3 transcription in human CD4+ T cells between -70 kbp to -1 bp from the TIM-3 transcription start site. Given that regulatory DNA regions show nucleotide sequence homology between species, four conserved non-coding sequences (CNS -29, CNS -53, CNS -56 and CNS -63) were analyzed by comparison human TIM-3 upstream DNA sequences with mouse and cow counterparts. Only at CNS -63, H3K4 dimethylation that is associated with transcriptional activation was observed in Jurkat T cells and CD4+ T cells isolated from human blood. Further, CNS -63 increased luciferase reporter gene expression downstream of TIM-3 minimal promoter. Enhancer activity of CNS -63 was abrogated by mutation of putative NF-κB binding sites between -63423 and -63433. These results indicate that TIM-3 CNS -63 may regulate TIM-3 transcription in human CD4+ T cells.","abstract_has_math":false,"creators":["Komori, Kuniharu"],"institution":null,"degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":["박, 선","대학원 의생명과학과","201124340"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2013,"date_issued":"2013-12-13T01:54:01Z","date_published":"2013-12-13T01:54:01Z","updated_at":"2026-07-24T00:51:42Z","subjects":["TIM-3","Histone H3 methylatio","Transcriptionregulatio"],"languages":["ko"],"rights":[],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier","label":"Identifier","values":["http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000013572","000000013572"],"render_values":[{"text":"http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000013572","href":"http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000013572","code":true},{"text":"000000013572","href":null,"code":true}]}]},"links":{"outbound_url":"http://repository.ajou.ac.kr/handle/201003/8590","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["박, 선","대학원 의생명과학과","201124340","Komori, Kuniharu"]},{"key":"dc:creator","label":"Author","values":["Komori, Kuniharu"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2013-12-13T01:54:01Z","2013"]},{"key":"dc:type","label":"Dc Type","values":["Thesis","Theses"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["TIM-3","Histone H3 methylatio","Transcriptionregulatio"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["ko"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["http://repository.ajou.ac.kr/handle/201003/8590","http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000013572","000000013572"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["The T cell immunoglobulin domain and mucin domain-3 (TIM-3) was discovered in Th1 and has been known as a T cell regulatory molecule. TIM-3 expression is induced by T cell activation through mechanism not yet well-investigated. In the study, I wanted to find DNA region involved in regulation of TIM-3 transcription in human CD4+ T cells between -70 kbp to -1 bp from the TIM-3 transcription start site. Given that regulatory DNA regions show nucleotide sequence homology between species, four conserved non-coding sequences (CNS -29, CNS -53, CNS -56 and CNS -63) were analyzed by comparison human TIM-3 upstream DNA sequences with mouse and cow counterparts. Only at CNS -63, H3K4 dimethylation that is associated with transcriptional activation was observed in Jurkat T cells and CD4+ T cells isolated from human blood. Further, CNS -63 increased luciferase reporter gene expression downstream of TIM-3 minimal promoter. Enhancer activity of CNS -63 was abrogated by mutation of putative NF-κB binding sites between -63423 and -63433. These results indicate that TIM-3 CNS -63 may regulate TIM-3 transcription in human CD4+ T cells.","The T cell immunoglobulin domain and mucin domain-3 (TIM-3)는 Th1의 세포표면에서 처음 발견되었고, T 세포의 면역반응을 억제하는 조절인자로 알려져 왔다. 하지만 TIM-3의 발현 조절 기작에 대해서는 알려져 있지 않다. 본 연구에서는 CD4+ T 세포에서 TIM-3 유전자 발현을 조절하는 enhancer를 조사하기 위해 먼저 TIM-3를 발현한다고 알려진 사람, 생쥐 그리고 소에서 TIM-3 유전자의 5' upstream 부분 (-70 Kb)의 염기서열을 분석하여 상동성이 80% 이상인 네 군데의 CNS -29 (-29602 bp ∼ -30324 bp), CNS -53 (-53882 bp ∼ -53934 bp), CNS -56 (-56508 bp ∼ -57191 bp) 그리고 CNS -63 (-63360 bp ∼ -63467 bp) 부분을 확인하였다. 그리고 Jurkat T 세포에서 TIM-3의 발현을 유도하는 phorbol 12-myristate 13-acetate (PMA)와 ionomycin 자극으로 인한 T 세포의 활성화에 따른 히스톤 메틸화 상태와 정상 세포에서 TIM-3 발현 유무에 따른 히스톤 메틸화 상태를 ChIP assay를 통해 분석하였다. CNS -63에서 H3K4 dimethylation (전사활성에 관여)이 관찰 되었고, H3K9 trimethylation (전사 억제에 관여)의 변화는 관찰되지 않았다. CNS -63의 enhancer activity를 luciferase reporter assay를 통해 분석하였다. T3U290 (CNS -63)에 의한 luciferase 활성이 T3U(290)과 자극이 없을 때 유사하게 나타났으며 PMA와 ionomycin에 의한 자극이 있을 때 luciferase 활성이 1.45배 증가하였다. CNS -63의 NF-κB 결합 예상 부위를 변이 시킨 T3U290 (Mut 2)에 의한 luciferase 활성이 PMA와 ionomycin에 의한 자극이 있을 때 T3U290 (CNS -63)보다 감소하였다. 이러한 결과로 보았을 때 CNS -63이 활성 자극에 의한 CD4+ T 세포에서 TIM-3 발현 증가에 관련 될 것이고 본 연구의 결과는 TIM-3 발현 조절을 이해하는데 중요한 기초를 제공할 것이다.","국문요약 ⅰ 차례 ⅲ 그림차례 ⅴ 표 차례 ⅵ Ⅰ.서론 1 Ⅱ.재료 및 방법 5 A.Conservedsequence(CNS)분석 5 B.세포배양 5 C.NaiveCD4+T 세포와 CD4+T 세포의 분리 5 D.Flow cytometry 6 E.CD4+T 세포 활성화 7 F.ChromatinImmuneprecipitation(ChIP)assay 7 G.Luciferasereportervector제작 8 H.Luciferaserepotorassay 9 Ⅲ.결과 12 A.TIM-3유전자 5'upstream region의 conservedsequence 12 B.CD4+T 세포에서 히스톤 H3의 메틸화 분석 12 C.TIM-3유전자 5'말단의 CNS enhancer역할 분석 18 Ⅳ.고찰 23 Ⅴ.결론 26 참고문헌 27 ABSTRACT 34","Master"]},{"key":"dc:format","label":"Dc Format","values":["text/plain"]},{"key":"dc:title","label":"Title","values":["Upstream conserved sequence element that regulates TIM-3 induction in CD4+ T cell","CD4+ T 세포에서 TIM-3 발현 유도에 관여하는 TIM-3 5' 말단의 종간 상동 염기서열"]}]}],"canonical_facts":{"dc:contributor":["박, 선","대학원 의생명과학과","201124340","Komori, Kuniharu"],"dc:creator":["Komori, Kuniharu"],"dc:date":["2013-12-13T01:54:01Z","2013"],"dc:description":["The T cell immunoglobulin domain and mucin domain-3 (TIM-3) was discovered in Th1 and has been known as a T cell regulatory molecule. TIM-3 expression is induced by T cell activation through mechanism not yet well-investigated. In the study, I wanted to find DNA region involved in regulation of TIM-3 transcription in human CD4+ T cells between -70 kbp to -1 bp from the TIM-3 transcription start site. Given that regulatory DNA regions show nucleotide sequence homology between species, four conserved non-coding sequences (CNS -29, CNS -53, CNS -56 and CNS -63) were analyzed by comparison human TIM-3 upstream DNA sequences with mouse and cow counterparts. Only at CNS -63, H3K4 dimethylation that is associated with transcriptional activation was observed in Jurkat T cells and CD4+ T cells isolated from human blood. Further, CNS -63 increased luciferase reporter gene expression downstream of TIM-3 minimal promoter. Enhancer activity of CNS -63 was abrogated by mutation of putative NF-κB binding sites between -63423 and -63433. These results indicate that TIM-3 CNS -63 may regulate TIM-3 transcription in human CD4+ T cells.","The T cell immunoglobulin domain and mucin domain-3 (TIM-3)는 Th1의 세포표면에서 처음 발견되었고, T 세포의 면역반응을 억제하는 조절인자로 알려져 왔다. 하지만 TIM-3의 발현 조절 기작에 대해서는 알려져 있지 않다. 본 연구에서는 CD4+ T 세포에서 TIM-3 유전자 발현을 조절하는 enhancer를 조사하기 위해 먼저 TIM-3를 발현한다고 알려진 사람, 생쥐 그리고 소에서 TIM-3 유전자의 5' upstream 부분 (-70 Kb)의 염기서열을 분석하여 상동성이 80% 이상인 네 군데의 CNS -29 (-29602 bp ∼ -30324 bp), CNS -53 (-53882 bp ∼ -53934 bp), CNS -56 (-56508 bp ∼ -57191 bp) 그리고 CNS -63 (-63360 bp ∼ -63467 bp) 부분을 확인하였다. 그리고 Jurkat T 세포에서 TIM-3의 발현을 유도하는 phorbol 12-myristate 13-acetate (PMA)와 ionomycin 자극으로 인한 T 세포의 활성화에 따른 히스톤 메틸화 상태와 정상 세포에서 TIM-3 발현 유무에 따른 히스톤 메틸화 상태를 ChIP assay를 통해 분석하였다. CNS -63에서 H3K4 dimethylation (전사활성에 관여)이 관찰 되었고, H3K9 trimethylation (전사 억제에 관여)의 변화는 관찰되지 않았다. CNS -63의 enhancer activity를 luciferase reporter assay를 통해 분석하였다. T3U290 (CNS -63)에 의한 luciferase 활성이 T3U(290)과 자극이 없을 때 유사하게 나타났으며 PMA와 ionomycin에 의한 자극이 있을 때 luciferase 활성이 1.45배 증가하였다. CNS -63의 NF-κB 결합 예상 부위를 변이 시킨 T3U290 (Mut 2)에 의한 luciferase 활성이 PMA와 ionomycin에 의한 자극이 있을 때 T3U290 (CNS -63)보다 감소하였다. 이러한 결과로 보았을 때 CNS -63이 활성 자극에 의한 CD4+ T 세포에서 TIM-3 발현 증가에 관련 될 것이고 본 연구의 결과는 TIM-3 발현 조절을 이해하는데 중요한 기초를 제공할 것이다.","국문요약 ⅰ 차례 ⅲ 그림차례 ⅴ 표 차례 ⅵ Ⅰ.서론 1 Ⅱ.재료 및 방법 5 A.Conservedsequence(CNS)분석 5 B.세포배양 5 C.NaiveCD4+T 세포와 CD4+T 세포의 분리 5 D.Flow cytometry 6 E.CD4+T 세포 활성화 7 F.ChromatinImmuneprecipitation(ChIP)assay 7 G.Luciferasereportervector제작 8 H.Luciferaserepotorassay 9 Ⅲ.결과 12 A.TIM-3유전자 5'upstream region의 conservedsequence 12 B.CD4+T 세포에서 히스톤 H3의 메틸화 분석 12 C.TIM-3유전자 5'말단의 CNS enhancer역할 분석 18 Ⅳ.고찰 23 Ⅴ.결론 26 참고문헌 27 ABSTRACT 34","Master"],"dc:format":["text/plain"],"dc:identifier":["http://repository.ajou.ac.kr/handle/201003/8590","http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000013572","000000013572"],"dc:language":["ko"],"dc:subject":["TIM-3","Histone H3 methylatio","Transcriptionregulatio"],"dc:title":["Upstream conserved sequence element that regulates TIM-3 induction in CD4+ T cell","CD4+ T 세포에서 TIM-3 발현 유도에 관여하는 TIM-3 5' 말단의 종간 상동 염기서열"],"dc:type":["Thesis","Theses"]},"updated_at":"2026-07-24T00:51:42Z"}