{"created":"2023-07-27T07:01:01.525807+00:00","id":61157,"links":{},"metadata":{"_buckets":{"deposit":"fdb97724-02ed-4158-85d5-378b9b362b83"},"_deposit":{"created_by":18,"id":"61157","owners":[18],"pid":{"revision_id":0,"type":"depid","value":"61157"},"status":"published"},"_oai":{"id":"oai:kanazawa-u.repo.nii.ac.jp:00061157","sets":["2812:2813:2841"]},"author_link":["21546","22481"],"item_9_biblio_info_8":{"attribute_name":"書誌情報","attribute_value_mlt":[{"bibliographicIssueDates":{"bibliographicIssueDate":"1994-03-23","bibliographicIssueDateType":"Issued"},"bibliographicPageStart":"3p.","bibliographicVolumeNumber":"1991 – 1992","bibliographic_titles":[{"bibliographic_title":"平成4(1992)年度 科学研究費補助金 国際学術研究 研究成果報告書概要"},{"bibliographic_title":"1992 Fiscal Year Final Research Report Summary","bibliographic_titleLang":"en"}]}]},"item_9_creator_33":{"attribute_name":"著者別表示","attribute_type":"creator","attribute_value_mlt":[{"creatorNames":[{}],"nameIdentifiers":[{},{}]}]},"item_9_description_21":{"attribute_name":"抄録","attribute_value_mlt":[{"subitem_description":"1987-1989年本研究代表者らは,米国National Cancer Instituteにて活性化ヒト末梢血単核球由来好中球,Tリンパ球,単球走化性サイトカインの精製ならびにそれらの遺伝子クローニングに成功した。好中球走化性因子とTリンパ球走化性因子はアミノ酸配列上同一であることが判明し,インターロイキン8(IL8)と命名した。単球走化性因子はIL8とアミノ酸配列上21%同一であり,単球活性化作用も有することよりmonocyte chemotactic and activating factor(MCAF)と命名した。両サイトカインは8kDaへパリン結合性で塩基性の強い白血球走化性サイトカイン(Chemokine)ファミリーに属する。本共同研究に於ては,IL8とMCAFの基礎ならびに前臨床実験として以下のような研究課題に取り組んだ。\n1)IL8,MCAFの構造と活性中心の決定---ヒトIL8を大腸菌で大量発現の後精製し,NMRとX線結晶回析によりIL8の3次元構造モデルを作製した。その結果,IL8は向かい合う形で水素結合をとうして二量体を形成,本体は二重のS-S結合を持つ三重鎖のベータシート構造をとりC-末端側は完全なアルファヘリックス構造をとることが判明した。IL8/MCAFの欠失変異体,キメラ分子を遺伝子組み替え法により大腸菌ならびにCHOホ乳動物細胞で発現し白血球走化性活性と受容体結合性を検討した。その結果,活性中心/受容体結合特異性決定部位はIL8分子ではHis33を中心にした領域にあり,C-末端部位は分子の安定化に寄与することが判明した。\n2)IL8,MCAF受容体の性状決定---ヒトIL8受容体は分子量60,000の糖蛋白で好中球上には20,000個/細胞表現されており,平衡定数は1nMと算定された。MCAF受容体は,分子量40,000で単核球上には13,000個/細胞表現されており,結合平衡定数は25nMと算定された。IL8受容体表現はIL8のみならずTNF,PMAなどによりdynamicに制御されており,また,MCAF受容体は非常に温度依存性の結合を示すことが解かった。IL8受容体cDNAは米国の他のグループによりクローニングされたが,我々はIL8受容体ファミリーに属する数個の新しいcDNAをクローニングした。\n3)IL8遺伝子発現機構の解析---ヒトIL8cDNAのクローニングの後,種々の細胞が,IL1,TNF,enodotoxin,exotoxin,viralproteins,重金属,活性酸素発生物質など様々の刺激によりIL8を産生することを明確にした。また,IL8の産生が,glucocorticoid,vitamin D3,lipooxygenase inhibitors,FK506などにより抑制されることも明確にした。ヒトIL8染色体遺伝子をクローニングし,IL8遺伝子5′-上流域のIL8産生制御物質反応性エンハンサー/サプレッサーの性状を検索した。線維芽細胞,神経膠細胞では-94bpから-71bpに存在するC/EBP(NF-IL6)とNF-kB核蛋白結合部位が種々の刺激物質共通に反応するエンハンサーとして働き,Tリンパ球,肝細胞,胃癌細胞においては-126bpから-120bpに存在するAP-1領域と-80bpから-71bpに存在するNFkB核蛋白結合部位の相乗的作用によりIL8遺伝子の転写が亢進することがわかった。glucocorticoid,FK506などの産生抑制剤もこれらのIL8エンハンサー結合蛋白を修飾することによりIL8遺伝子転写を抑制していることもわかった。\n4)IL8,MCAFの病態生理作用の確立ならびに臨床応用の可能性の検索---IL8がRA,炎症性大腸炎,皮膚乾癬症,腎炎,尿路感染症など種々のヒト炎症疾患で産生されることが判明しIL8測定が炎症の良いマーカーになることがわかった。IL8の急性炎症時の好中球浸潤に於ける本質的関与に関してウサギを用いて関節炎,皮膚炎,肺再潅流症候群モデルにて証明することができた。IL8,MCAFcDNA導入実験腫瘍にては有意な抗腫瘍効果を観た。現在,IL8関連サイトカイン遺伝子導入腫瘍の成着率,転移能について評価中である。MCAFは,in vitroにてはマウスマクロファージをprimingしてより強い抗腫瘍効果を示すことがわかった。MCAFはまた,マウス実験にて種々の細菌に対して前投与にて抗菌作用を有することが判明した。今のところIL8,MCAFの造血作用は証明できない。","subitem_description_type":"Abstract"},{"subitem_description":"BASIC AND PRECLINICAL EXPERIMENTS OF IL 8 AND MCAF\nNovel chemotactic cytokines, IL 8 and MCAF were identified, purified, and molecularly cloned by us at National Cancer Institute, USA in 1987-1989, belong a family of emerging chemotactic cytokine family, CHEMOKINE. In this international collabolative research, the following projects were performed.\n1)Structural analysis of IL 8/MCAF and determining their active site(s)-----Human IL 8 was expressed in E coli and purified to homogeneity in a large scale. The structure of IL 8 was analyzed by both NMR and X-ray crystallography. The proposed model shows that IL 8 exists as dimer through hydrogen bonding and two symmetry-related anti-parallel a-helices, 24A long and separated by 14A, lie on top of a six-stranded anti-parallel b-sheet platform. The active site of IL 8 was presumed to be the region surrounding His at 33 as well as N-terminal region based on the mutation studies and structural analyses.\n2)Structure of the receptors for IL 8 and MCAF-----The MW of the receptors for IL 8 on human nutrophils was estimated to be 60,000 with Kd of lnM. The MW of the receptors for MCAF on human monocytes was estimated to be 40,000 with Kd of 25nM. Although the cDNA for IL 8 was cloned by other groups, we have cloned several novel cDNA which belong to the family of chemotactic cytokine receptors.\n3)Molecular analysis of the regulation of the production of IL 8-----After cDNA cloning of human IL 8, we revealed the induction of the production of IL 8 by various types of cells after stimulating with IL 1, TNF, endotoxin, exotoxin, viral proteins, heavy metals, and superoxide generating substances. We also showed the suppression of the production of IL 8 by glucocorticoids, vitamin D3, lipooxygenase inhibitors, and FK506. We have determined the enhancer of the human IL 8 gene to be consisted of AP-1+NFKB. IL 8 production suppressive agents seem to inhibit the activation of nuclear factors which bind to these regions.\n4)Establishment of pathophysiological roles of IL 8/MCAF and examining possible clinical application of IL 8/MCAF-----We have revealed the production of IL 8/MCAF in various human inflammatory diseases including ulcerative colitis, psoriasis, glomerulonephritis, urinary tract infection, and arthritis. Essential involvement of IL 8 in recruiting neutrophils in acute inflammation models in rabbits, such as dermatitis, arthritis, and lung reperfusion was established using monoclonal neutralizing antibody against rabbit IL 8. IL 8/MCAF cDNA transfection into tumor cells showed significant anti-tumor effect of these cytokines in mice. MCAF is further shown to prime murine macrophages to be tumor cytocidal and also have anti-infectious activity in mice. Possible hematopoietic activity of these cytokines was not proved either in vitro or in vivo.","subitem_description_type":"Abstract"}]},"item_9_description_22":{"attribute_name":"内容記述","attribute_value_mlt":[{"subitem_description":"研究課題/領域番号:03044066, 研究期間(年度):1991 – 1992","subitem_description_type":"Other"},{"subitem_description":"出典：研究課題「新しいサイトカイン.IL8とMCAFの基礎ならびに前臨床実験」課題番号03044066\n（KAKEN：科学研究費助成事業データベース（国立情報学研究所）） \n（https://kaken.nii.ac.jp/ja/report/KAKENHI-PROJECT-03044066/030440661992kenkyu_seika_hokoku_gaiyo/）を加工して作成","subitem_description_type":"Other"}]},"item_9_description_5":{"attribute_name":"提供者所属","attribute_value_mlt":[{"subitem_description":"金沢大学がん研究所","subitem_description_type":"Other"}]},"item_9_identifier_registration":{"attribute_name":"ID登録","attribute_value_mlt":[{"subitem_identifier_reg_text":"10.24517/00067401","subitem_identifier_reg_type":"JaLC"}]},"item_9_relation_28":{"attribute_name":"関連URI","attribute_value_mlt":[{"subitem_relation_name":[{"subitem_relation_name_text":"https://kaken.nii.ac.jp/search/?qm=50222427"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://kaken.nii.ac.jp/search/?qm=50222427","subitem_relation_type_select":"URI"}},{"subitem_relation_name":[{"subitem_relation_name_text":"https://kaken.nii.ac.jp/ja/grant/KAKENHI-PROJECT-03044066/"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://kaken.nii.ac.jp/ja/grant/KAKENHI-PROJECT-03044066/","subitem_relation_type_select":"URI"}},{"subitem_relation_name":[{"subitem_relation_name_text":"https://kaken.nii.ac.jp/ja/report/KAKENHI-PROJECT-03044066/030440661992kenkyu_seika_hokoku_gaiyo/"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://kaken.nii.ac.jp/ja/report/KAKENHI-PROJECT-03044066/030440661992kenkyu_seika_hokoku_gaiyo/","subitem_relation_type_select":"URI"}}]},"item_9_version_type_25":{"attribute_name":"著者版フラグ","attribute_value_mlt":[{"subitem_version_resource":"http://purl.org/coar/version/c_ab4af688f83e57aa","subitem_version_type":"AM"}]},"item_files":{"attribute_name":"ファイル情報","attribute_type":"file","attribute_value_mlt":[{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2022-10-24"}],"displaytype":"detail","filename":"CA-PR-MATSUSHIMA-K-kanen 1994-3p.pdf","filesize":[{"value":"109.1 kB"}],"format":"application/pdf","licensetype":"license_11","mimetype":"application/pdf","url":{"label":"CA-PR-MATSUSHIMA-K-kanen 1994-3p.pdf","url":"https://kanazawa-u.repo.nii.ac.jp/record/61157/files/CA-PR-MATSUSHIMA-K-kanen 1994-3p.pdf"},"version_id":"ab0d3c7d-1089-4b44-a565-daa95f5086d0"}]},"item_language":{"attribute_name":"言語","attribute_value_mlt":[{"subitem_language":"jpn"}]},"item_resource_type":{"attribute_name":"資源タイプ","attribute_value_mlt":[{"resourcetype":"research report","resourceuri":"http://purl.org/coar/resource_type/c_18ws"}]},"item_title":"新しいサイトカイン.IL8とMCAFの基礎ならびに前臨床実験","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"新しいサイトカイン.IL8とMCAFの基礎ならびに前臨床実験"},{"subitem_title":"Basic and preclinical experiments of IL 8 and MCAF","subitem_title_language":"en"}]},"item_type_id":"9","owner":"18","path":["2841"],"pubdate":{"attribute_name":"公開日","attribute_value":"2022-10-24"},"publish_date":"2022-10-24","publish_status":"0","recid":"61157","relation_version_is_last":true,"title":["新しいサイトカイン.IL8とMCAFの基礎ならびに前臨床実験"],"weko_creator_id":"18","weko_shared_id":-1},"updated":"2023-07-27T12:04:46.096974+00:00"}