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1型単純ヘルペス実質性角膜炎の実験モデルによる免疫病理学的解析と治療の研究

1型单纯疱疹实质角膜炎实验模型的免疫病理学分析及治疗研究

基本信息

批准号：
61570851
负责人：
内田幸男
金额：
$ 1.54万
依托单位：
Tokyo Women's Medical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1986
资助国家：
日本
起止时间：
1986 至 1988
项目状态：
已结题

项目摘要

1.各種単純ヘルペス1型株 (HSV-1) を8週令BalB/cマウス角膜に接種して病変形成能を比較した。その結果CHR3株が最も良く実質炎を伴う角膜ヘルペスを惹起した。弱毒株Ska.KOS株では角膜上皮、実質共に炎症を惹起し難にかった。2.1の結果からCHR3株をBalb/cマウス角膜に接種した時の細胞性免疫反応と病変形成・感染防禦に関与するT細胞の解析をすすめた。その結果 (1) 局所 (耳前、頚下) リンパ節に感染7日後に誘導されるHSV特異的細胞障害性T細胞は脾のそれに比し約10倍防禦活性が高い。 (2) 感染局所リンパ節細胞を抗体+補体処理によりL_3T_4細胞および、Lyt_2細胞に分けたのちヌードマウスに移入して角膜HSV-1感染に対する防禦能をしらべると、L_3T_4及びLyt_2細胞ともに活性があるが、共に存在する時に最も強い防禦効果が得られる。 (3) 局所リンパ節細胞をin vitroで5日間培養し、その間に各種抗原刺激を与えたのちにヌードマウスに移入して各T細胞サブセットの活性誘導条件を検討した。UV不活化HSV-1を加えて培養するとL_3T_4細胞にのみ強い防禦活性がみられた。LiveHSV-1を加えると、L_3T_4、Lyt_2細胞ともに活性は認められなかった。Balb/c胎児線維芽細胞 (MEF) にHSV-1を感染し24時間後にγ線照射を行って細胞を不活化し抗原として用いるとL_3T_4、Lyt_2細胞の両者に活性がみとめられる。クロロキン処理γ線照射HSV-1感染MEFや、感染MEFをγ線照射後Anti-H-2d処理したものを抗原として用いるとLyt_2細胞に活性は認められない。Anti-lAd抗体処理ではL_3T_4細胞が誘導されなくなる。これらの事から、L_3T_4細胞の誘導にはIA+HSV-1抗原、Lyt_2細胞の誘導にはH+2+HSV-1抗原が必要であることがわかる。 (4) 各サブセットT細胞が単独で生体でそれぞれ活性化しうるか否かについてヌードマウスに正常Balb/cからのL_3T_4、Lyt_2細胞を移した後HSV-1を感染させて各細胞の活性を見ると両者が存在する時よく防禦能が誘導された。

1. All kinds of HSV-1 strains (8 strains) make it possible for BalB/c to transplant corneal disease. The results showed that the most benign inflammatory disease of CHR3 strain was accompanied by corneal inflammation. The attenuated strain Ska.KOS caused inflammation of corneal epithelium and corneal epithelium. Results CHR3 strain Balb/c-infected cornea were inoculated with chronically infective cellular immune reflux disease to form anti-infection antibody and T-cell cytometric assay. Results (1) after 7 days of infection, the cytotoxic T cells of HSV were about 10 times higher than those of the control group. (2) the antibody against HSV-1 infection in the cornea, the activity of L_3T_4 and Lyt_2 in the infected cornea were significantly higher than those in the control group. (2) the antibodies against HSV-1 infection in the cornea were significantly higher than those in the control group. (2) the antibody against HSV-1 infection in the cornea was significantly higher than that in the control group (P < 0.01). (3) all kinds of antigen stimuli and antigens were cultured and tested within 5 days in the cell in vitro of the local government. They were transferred into the T cells of each cell in the first place. UV does not activate HSV-1 to increase the concentration of L _ 3T_4 cells to increase the activity of anti-virus. Add LiveHSV-1, L_3T_4, and Lyt_2 cells to determine the activity of each cell. Balb/c fetal line maintenance germ cells (MEF) were infected with HSV-1 24 days later, and then gamma rays were irradiated after 24 hours of infection. Cell inactivated antigens were used by Elisa L _ 3T_4, and Lyt_2 cells were activated by Elisa. After HSV-1 was infected by γ-rays and infected with MEF by γ-rays, Anti-H-2d was infected by γ-rays, and the antigens were detected by immunocytochemistry. The Anti-lAd antibody cleaved the L_3T_4 cells to guide the immune response. It is necessary for L_3T_4 cells to direct IA+HSV-1 antigens and Lyt_2 cells to induce H+2+HSV-1 antigens. (4) the activity of Balb/c-1 positive cells in each cell cycle was different from that of normal HSV-1 cells, and the cell activity of Lyt_2-1 infected cells showed that there was an occasional protective response.