Analysis of biological roles of class II major histocompatibility antigen by anti-sense RNA

反义RNA分析II类主要组织相容性抗原的生物学作用

• 批准号: 62480163
• 负责人: TOKUHISA Takeshi
• 金额: $ 4.35万
• 依托单位: Kobe University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1987
• 资助国家: 日本
• 起止时间: 1987 至 1988
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-62480163/
• 关键词: class II major hitocompatibility antigen; anti-sense RNA; Gene transfection; Protoplast method; antigen presenting activity; クラスII主要組織適合抗原; Antisense RNA; プロトプラスト法; 抗原提示能

We have established an anti-sense RNA system which is capable of regulating the expression of the endogenous class II MHC (la) molecule in order to elusidate the role of la molecules on immune responses. The 100 base pairs of the l-A chain gene were constructed under the control of a metallothionein gene promoter in an anti-sense orientation. This anti-sense gene was taransfected into the la^+ B lymphoma cells. The amount of la molecules on the transfectants was specifically reduced to less than 5% of the control level. The rations of the amount of anti-sense and sense mRNA in the transfectants correlated with the amount of the la molecules, indicating that the duplex fomation of sense and anti-sense RNA inhibits translation of the sense mRNA. Antigen presenting cell activities of the transfectants to the la restricted T cells also correlated with the amount of la molecules.simultaneously, this anti-sense gene was microinjected into fertilized eggs of C57BL/6xSJL F1 mice to raise the transgenic mice. In these transgenic mice, 1)the amount of la molecules on the spleen cells was distinctly reduced, 2) anti-sense rna was detected in total RNA from spleen cells of the transgenic mice. We are examining immune abnormalities of these mice.

我们已经建立了一个反义RNA系统，该系统能够调节内源性II类MHC（Ia）分子的表达，以洗脱Ia分子在免疫应答中的作用。在反义方向的金属硫蛋白基因启动子的控制下构建100个碱基对的I-A链基因。将该反义基因转染到la^+ B淋巴瘤细胞中。转染子上的Ia分子的量特异性地降低至低于对照水平的5%。转染子中反义mRNA和正义mRNA的量的比率与Ia分子的量相关，表明正义RNA和反义RNA的双链体形成抑制正义mRNA的翻译。同时，将该反义基因显微注射到C57 BL/6xSJLF 1小鼠受精卵中，培育出转基因小鼠。在这些转基因小鼠中，1）脾细胞上的Ia分子的量明显减少，2）在转基因小鼠的脾细胞的总RNA中检测到反义RNA。我们正在检查这些小鼠的免疫异常。

项目成果

Hatano,M.; Aizawa,S.; Soejima,T.; Shigemoto,K.; Taniguch,M.; Tokuhisa,T.: "Specific regulation of class II MHC gene expression by anti-sense RNA." Int. Immunol.(1989)

波多野，M.；

Tokuhisa,T.;et al.: "Specific inhibition of the major histocompatibility complex(MHC)expression by anti-sense RNA." Proceedings of the 7th seminar on Science and.Technology"Molecular Biology", 249-262 (1988)

Tokuhisa,T. 等人：“反义 RNA 对主要组织相容性复合物 (MHC) 表达的特异性抑制。”

幡野雅彦 他: 日本免疫学会誌. 18. 752 (1988)

Masahiko Hatano 等人：日本免疫学会杂志 18. 752 (1988)。

Uchiyama,K.; Saito,H.; Tokuhisa,T.; Imai,K.; Taniguchi,M.: "High frequency of loss of human kappa light chain expression in mouse-human hetero hybridomas." Hybridoma. 6. 645-654 (1987)

内山，K.；

脇田和幸 他: 日本免疫学会誌. 18. 751 (1988)

Kazuyuki Wakita 等人：日本免疫学会杂志 18. 751 (1988)。