{"created":"2023-05-15T09:59:49.329897+00:00","id":4045,"links":{},"metadata":{"_buckets":{"deposit":"07b55a48-c985-48da-96fa-3f2643b7cafc"},"_deposit":{"created_by":9,"id":"4045","owner":"9","owners":[9],"pid":{"revision_id":0,"type":"depid","value":"4045"},"status":"published"},"_oai":{"id":"oai:miyazaki-u.repo.nii.ac.jp:00004045","sets":["74","74:59"]},"author_link":["21372"],"item_10006_description_7":{"attribute_name":"抄録","attribute_value_mlt":[{"subitem_description":"近年､非メチル化･CpGジヌクレオチド(ODN)を含んでいるDNA分子は､晴乳類においてすぐれた免疫賦活作用があることが報告されている｡\nしかし､これらのDNAモチーフが魚類の免疫系に及ぼす作用についてはほとんど研究されていない｡そこで本研究では､コイを用いてDNAのCpGモチーフがコイの白血球の活性に与える影響を調べた｡次にCpG-ODNs投与魚で発現する遺伝子をEST解析で調べ､さらにCpGモチーフがコイの白血球のサイトカイン(インターロイキン(IL)-1β､\nIL-10､TNF-α､CXC-およびCC-ケモカイン)遺伝子の発現に及ぼす影響についても検討を行った｡\nODNsは噂乳類で免疫賦活作用のあるモチーフとして知られている5種類のCpG-ODNs(A=TCCATGAEETTCCTGATGCT,B=GCTAGAGETTAAGSTT,\nC=ATCGACTCTCGAACG TTC C,D=GAA CCTTCCATG CTGTTC12G,E=GCTAGATGTTAGE⊆T)を合成し､本研究に用いた.\n試験管内でコイの腎臓の白血球をcpG-ODNsで刺激すると､CpG10DN BとCが食食能とNBT活性を有意に増強した｡さらにコイのリンパ球の幼若化を有意に促進した｡\n試験管内で有意な活性を示したCpG-ODN BまたはCをコイの筋肉内に1,10,100μg/kg/fish注射した｡その後､腎臓の白血球の活性化を貧食能とNBT法で､また血清中のリゾチーム活性についても測定を行った｡\nCpG-ODN BまたはCを注射したコイの腎臓の白血球の貧食能は､対照区に比べて有意な増加が確認された｡さらに白血球のNBT活性も有意に増加した｡また､血清中のリゾチーム活性もCpG-ODN BまたはCを注射した区において有意な増加が認められた｡\n次にこれらのコイの腎臓組織より作成したcDNAライブラリーより88クローンについてEST解析を行った結果､granulin2,CCAAT/enhancer binding protein, immunoglobulin \nheavy chain variable region, lectin, lysozyme C, interleukin-4 receptor alpha chain, \ncathepsin L preproprotein, CD9 protein, Granulin l などの免疫系に関与する遺伝子を分離した｡\n同様に処理したコイの腎臓由来白血球のサイトカイン遺伝子を経時的に半定量解析した結果､炎症性のサイトカインであるIL-1β､CXCおよびCC-ケモカイン遺伝子の発現量が増加した｡またCpG-ODN Cはlysozyme-C遺伝子の発現量を増加させた｡試験管内においてCpG-ODN BまたはCで処理されたコイの白血球からもIL-1β､CXCおよびCC-ケモカイン､TNF-α遺伝子の発現量増加が確認された.これらサイトカイン遺伝子の発現はCpG-ODNs処理後､速やかに高い発現量を示した.一方､抗炎症性サイトカインであるIL-10遺伝子の発現量については､CpG-ODNs処理による発現量の有意な変化は確認されなかった｡また､non-CPG-ODNsではサイトカイン遺伝子の発現量の増加は確認されなかった｡\nこれらの結果よりCpG-ODNモチーフはコイの免疫応答を増強する作用があることが示された。\nAn immunostimulant is a substance or action, which causes an innate immune response or increases an adaptive immune response. Bacterial DNA has long been known to be able to activate immune system, whereas mammalian is not. An important difference between bacterial and mammalian DNA is the suppression of unmethylated CpG motifs in mammalian DNA. Synthetic oligodeoxynucleotides containing unmethylated CpG motifs have been shown to mimic the effect of bacterial DNA in exerting immune responses.\nThe study mainly focused on the investigation of immunostimulatory effects of CpG oligodeoxynucleotides (CpG-ODNs) on the immune system of common carp (Cyprinus\ncarpio L.). The effects of a panel of synthetic oligodeoxynucleotides on the\nimmunocompetence cell activity in common carp were examined. In vitro addition of\nCpG-ODNs enhanced phagocytic function (reduction of nitroblue tetrazolium, phagocytosis) from stimulated kidney phagocytes. The CpG-ODNs also induced lymphocyte proliferation in the fish kidney cells. These results show that CpG-ODNs could potentially improve immunocompetence cell activity in fish through increased cell proliferation and functionality.\nBased on the results from in vitro study in which CpG-ODNs with the mice and fish motif\n(GA/AA)CGTI, were shown to stimulate phagocyte and lymphocyte activities in common carp head kidney leucocytes, in vivo studies were undertaken to examine the capacity of these CpG-ODNs to stimulate phagocytic and serum lysozyme activities in the carp leucocytes.\nIntraperitoneal injection of CpG-ODNs to fish daily, 3 days, resulted in enhanced responses of phagocytic and nitroblue tetrazolium (NBT) activities in kidney phagocytic cells. This activation of kidney cells was observed for at least 7 days post-injection. The serum lysozyme activity also increased in the fish treated with CpG-ODNs. These results indicated that unmethylated CpG DNA enhance the innate immune response of carp.\nAnalysis of expressed sequence tags of 88 clones from CpG-ODNs stimulated head kidney\nof carp was carried out. Several immune-related genes were identified during this analysis, including granulin 2, CCAAT/enhancer binding protein, immunoglobulin heavy chain variable region, lectin, lysozyme C, interleukin-4 receptor alpha chain, cathepsin L preproprotein, CD9\nprotein and Granulin 1. Using head kidney macrophages of common carp as an in vivo model, the effects of CpG-ODNs on expression of a number of immune-related genes in fish macrophages were investigated. CpG-ODNs augmented expression of interleukin (IL)-1 1β, CXC and CC-chemokines at 1, 5 and 7 days post-treatment. CpG-ODN also increased the Iysozyme-C gene expression at 7 days post-injection. Using in vitro approaches, CpG-ODNs were shown to up-regulate the expression of IL-lβ, CXC and CC-chemokines, and tumor necrosis factor (TNF)-a genes in the head kidney of carp. The high level expressions of these genes were seen at an earlier stage of CpG-ODNs stimulation. The IL-10 gene expressions were not significantly enhanced by CpG-ODNs and the non-CpG ODNs also failed to stimulate the expression of these cytokine genes.\nThese studies have shown that CpG-ODNs are effective immune stimulators for fish immune cells, suggesting that CpG-ODNs may be useful in enhancing the immune responses\nin veterinary applications for fish.","subitem_description_language":"ja","subitem_description_type":"Abstract"}]},"item_10006_textarea_22":{"attribute_name":"内容記述","attribute_value_mlt":[{"subitem_textarea_value":"鹿児島大学大学院連合農学研究科博士論文"},{"subitem_textarea_language":"ja"}]},"item_10006_version_type_18":{"attribute_name":"著者版フラグ","attribute_value_mlt":[{"subitem_version_resource":"http://purl.org/coar/version/c_970fb48d4fbd8a85","subitem_version_type":"VoR"}]},"item_creator":{"attribute_name":"著者","attribute_type":"creator","attribute_value_mlt":[{"creatorNames":[{"creatorName":"Tassakka, Asmi Citra Malina  A.R.","creatorNameLang":"en"}],"nameIdentifiers":[{}]}]},"item_files":{"attribute_name":"ファイル情報","attribute_type":"file","attribute_value_mlt":[{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2020-06-21"}],"displaytype":"detail","filename":"DT_A.R.TassakkaAsmiCitraMalina.pdf","filesize":[{"value":"6.3 MB"}],"format":"application/pdf","licensetype":"license_note","mimetype":"application/pdf","url":{"label":"DT_A.R.TassakkaAsmiCitraMalina.pdf","url":"https://miyazaki-u.repo.nii.ac.jp/record/4045/files/DT_A.R.TassakkaAsmiCitraMalina.pdf"},"version_id":"352c54e8-077d-4ebe-b5d9-9945dbd1118d"}]},"item_language":{"attribute_name":"言語","attribute_value_mlt":[{"subitem_language":"eng"}]},"item_resource_type":{"attribute_name":"資源タイプ","attribute_value_mlt":[{"resourcetype":"thesis","resourceuri":"http://purl.org/coar/resource_type/c_46ec"}]},"item_title":"DNAのCpGモチーフはコイの非特異免疫応答を活性化させるか","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"DNAのCpGモチーフはコイの非特異免疫応答を活性化させるか","subitem_title_language":"ja"},{"subitem_title":"CpG oligodeozynucleotides stimullate the immune system of common carp (Cyprinus carpio L.)","subitem_title_language":"en"}]},"item_type_id":"10006","owner":"9","path":["74","59"],"pubdate":{"attribute_name":"PubDate","attribute_value":"2007-11-26"},"publish_date":"2007-11-26","publish_status":"0","recid":"4045","relation_version_is_last":true,"title":["DNAのCpGモチーフはコイの非特異免疫応答を活性化させるか"],"weko_creator_id":"9","weko_shared_id":-1},"updated":"2023-07-30T23:00:47.514666+00:00"}