1998 年度実績報告書

ヒトCD46高発現トランスジェニック動物の開発

研究課題

研究課題/領域番号	10878157
研究機関	大阪府立成人病センター研究所
研究代表者	瀬谷司大阪府立成人病センター研究所, 第6部, 部長 (10301805)
研究分担者	長沢滋治北海道大学, 大学院・薬学系研究科, 教授 (70029958) 松本美佐子大阪府立成人病センター研究所, 第6部, 主任研究員
キーワード	ヒトCD46(membrane cofactor protein) / トランススジェニックマウス / 麻疹ウイルス / 補体制御因子 / 異種移植 / 3′-UTサイレンサー
研究概要	ヒトCD46を高発現するマウス作製のために以下のことを明らかにした。1.CD46の転写・翻訳の調節機構:ヒトCD46は4kb ubiquitous-messageと1.5kb testis-specific messageから成る。4kb ubiquitous message(L-form)のcDNAをマウス細胞に強制発現させてもヒトやサル細胞のようにCD46proteinは高発現しない。2.CD46のSCR1,SCR2が麻疹ウィルスレセプターとして機能する。SCR2-4は補体制御活性に必須なドメインである。従って、SCR1を削除したmutantは麻疹ウィルスレセプターとして機能せずに補体制御機能のみを発揮する。このことをブタ血管内皮細胞を用いて検証した。3.[CD46は免疫系を抑制するレセプターとしても機能しうる。シグナル伝達に関与するcytoplasmatic tailのsequenceを削除する必要がある。]この点は現在試行中である。 Preliminary trailで、L-form cDNAをトランスジーンしたがCD46高発現マウスは得られなかった。Constructは上記のごとく完成したのでCD46の高発現マウスを目指してさらに継続試行していく予定である。ヒトCD46は細胞を自己補体から守る免疫制御因子であり、かつ麻疹ウィルスレセプターとしても機能する。本研究でCD46高発現のマウスが得られれば、麻疹感染・異種移植の動物レベルでの解析に道が開かれる。もしCD46高発現のfounderが得られれば、ウィルス感染・移植の解析実験をより大きい規模で計画する予定である。

研究成果

(19件)

すべてその他

すべて文献書誌 (19件)

[文献書誌] Mikata.S: "Regulation of human complement-mediated swine endothelial cell lysis by surface-bound fonn of human C46-binding protein" Transplantation. 65. 363-368 (1998)
[文献書誌] Hatanaka,M.: "Homodimerization of a GPI-anchored complement regulatory protein CD59 on the surface of Hela and CD59-transfected CHO cells." J.Biochem.123. 579-586 (1998)
[文献書誌] Hara T.: "Post-translational modification and intracellular localization of a spl product of CD46 cloned from the human testis: Role of the intracellular domains in O-glycosylation" Immunology. 93. 546-555 (1998)
[文献書誌] Mikata.S: "New Approach to prevent complement activation on xenograffs:effects of a surface-bound of human C46-binding prortein" Transplant.Proc.30. 65-66 (1998)
[文献書誌] Matsumoto M.: "Structural and functional properties of complement-activating protein M161Ag a Mycoplasma fermentans gene products that induces cytokine production by human monocytes" J.Biol.Chem. 273. 12407-12414 (1998)
[文献書誌] Sakata H.: "A Japanese wild-type measles virus strain inducing predominarst early down-regulation of CD46." Biol.Pharmac.Bull.21. 1121-1127 (1998)
[文献書誌] Seya T: "CD46(membrane cofactor protein (MCP) of complement,measles virus receptor)structural and functional clivergence among speces(Review)" Int.J.Mol.Med.1. 809-816 (1998)
[文献書誌] Mikata S: "A monomeric human C46-binding protein (C46p) more efficiently inactivates C3b than natural C46p:Participation of C-terminal domains in factor-I." Mole.Immunol.35. 537-544 (1998)
[文献書誌] Saeki Y: "Phosphatidlylinositol 3 kinose inhibitors block matrix adhesion and experimental metastasis of high metastatic mouse hepatoma clones" J.Biochem. 124. 1020-1025 (1998)
[文献書誌] Hatanaka M: "Expression of caveolin-I in human T cell leukemia cell lines" Biochem.Biophys.Res.Commun. 253. 382-387 (1998)
[文献書誌] Begum N.A.: "A BCG-CWS-regulated gene in human monocytes by differential display: Identification of ahuman homologue of chicken MD-1" Biochem.Biophys.Res.Commun.(印刷中).
[文献書誌] Kawano M: "Elevated serum levels of soluble membrane cofactor protein(CD46 MCP)in patients with systemic lupus erythematosus." Clin.Exp.Immunol.(印刷中).
[文献書誌] Kurita M: "Antibody-independent classical complement pathway activation and homologous C3 deposition on human xeroderma cell lines" Clin.Exp.Immunol.(印刷中).
[文献書誌] Seya T: "Innate immune therapy for cancer,lessons from M161Ag,a mycoplasma product" Human Gene Therapy ed.by N.Habib IBC press,UKpp. (印刷中).
[文献書誌] Seya T: "Molecular remodeling of complement regulatory proteins for xenotransplantation" Immunopharmacology. (印刷中).
[文献書誌] Murakami Y: "Effect of mutations at the residues R25.D27,P69 and N70 of B95a-Map on receptor activities for the measles virus Nagahata wild-type strain and CAM vaccine strain" Int.J.Mol.Med.3. 25-32 (1999)
[文献書誌] Matsumoto M: "M161Ag is a potent cytokine inducer with complement activating function (Review)" Int.J.Mdec.Med.3. 291-295 (1999)
[文献書誌] Seya T: "Innate immune therapy for cancer:seeking for effective molecules for activation of the innate immune system" Human Gene Therapy ed.by N.Habib,Plenum Press USA pp. (印刷中).
[文献書誌] Seya T: "Human membrane cofactor protein of complement (CD46):multiple isoforms and functions" Int.J.Biochem.Cell.Biol. (印刷中).

1998 年度 実績報告書

ヒトCD46高発現トランスジェニック動物の開発

研究代表者

瀬谷 司 大阪府立成人病センター研究所, 第6部, 部長 (10301805)

研究成果

[文献書誌] Mikata.S: "Regulation of human complement-mediated swine endothelial cell lysis by surface-bound fonn of human C46-binding protein" Transplantation. 65. 363-368 (1998)

[文献書誌] Hatanaka,M.: "Homodimerization of a GPI-anchored complement regulatory protein CD59 on the surface of Hela and CD59-transfected CHO cells." J.Biochem.123. 579-586 (1998)

[文献書誌] Hara T.: "Post-translational modification and intracellular localization of a spl product of CD46 cloned from the human testis: Role of the intracellular domains in O-glycosylation" Immunology. 93. 546-555 (1998)

[文献書誌] Mikata.S: "New Approach to prevent complement activation on xenograffs:effects of a surface-bound of human C46-binding prortein" Transplant.Proc.30. 65-66 (1998)

[文献書誌] Matsumoto M.: "Structural and functional properties of complement-activating protein M161Ag a Mycoplasma fermentans gene products that induces cytokine production by human monocytes" J.Biol.Chem. 273. 12407-12414 (1998)

[文献書誌] Sakata H.: "A Japanese wild-type measles virus strain inducing predominarst early down-regulation of CD46." Biol.Pharmac.Bull.21. 1121-1127 (1998)

[文献書誌] Seya T: "CD46(membrane cofactor protein (MCP) of complement,measles virus receptor)structural and functional clivergence among speces(Review)" Int.J.Mol.Med.1. 809-816 (1998)

[文献書誌] Mikata S: "A monomeric human C46-binding protein (C46p) more efficiently inactivates C3b than natural C46p:Participation of C-terminal domains in factor-I." Mole.Immunol.35. 537-544 (1998)

[文献書誌] Saeki Y: "Phosphatidlylinositol 3 kinose inhibitors block matrix adhesion and experimental metastasis of high metastatic mouse hepatoma clones" J.Biochem. 124. 1020-1025 (1998)

[文献書誌] Hatanaka M: "Expression of caveolin-I in human T cell leukemia cell lines" Biochem.Biophys.Res.Commun. 253. 382-387 (1998)

[文献書誌] Begum N.A.: "A BCG-CWS-regulated gene in human monocytes by differential display: Identification of ahuman homologue of chicken MD-1" Biochem.Biophys.Res.Commun.(印刷中).

[文献書誌] Kawano M: "Elevated serum levels of soluble membrane cofactor protein(CD46 MCP)in patients with systemic lupus erythematosus." Clin.Exp.Immunol.(印刷中).

[文献書誌] Kurita M: "Antibody-independent classical complement pathway activation and homologous C3 deposition on human xeroderma cell lines" Clin.Exp.Immunol.(印刷中).

[文献書誌] Seya T: "Innate immune therapy for cancer,lessons from M161Ag,a mycoplasma product" Human Gene Therapy ed.by N.Habib IBC press,UKpp. (印刷中).

[文献書誌] Seya T: "Molecular remodeling of complement regulatory proteins for xenotransplantation" Immunopharmacology. (印刷中).

[文献書誌] Murakami Y: "Effect of mutations at the residues R25.D27,P69 and N70 of B95a-Map on receptor activities for the measles virus Nagahata wild-type strain and CAM vaccine strain" Int.J.Mol.Med.3. 25-32 (1999)

[文献書誌] Matsumoto M: "M161Ag is a potent cytokine inducer with complement activating function (Review)" Int.J.Mdec.Med.3. 291-295 (1999)

[文献書誌] Seya T: "Innate immune therapy for cancer:seeking for effective molecules for activation of the innate immune system" Human Gene Therapy ed.by N.Habib,Plenum Press USA pp. (印刷中).

[文献書誌] Seya T: "Human membrane cofactor protein of complement (CD46):multiple isoforms and functions" Int.J.Biochem.Cell.Biol. (印刷中).

1998 年度実績報告書

瀬谷司大阪府立成人病センター研究所, 第6部, 部長 (10301805)