Regulation and analysis of systemic and local effect of TNF

TNF全身和局部效应的调控与分析

• 批准号: 04454189
• 负责人: HIGUCHI Yasunori
• 金额: $ 3.97万
• 依托单位: Oita Medical 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: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04454189/
• 关键词: CD14; TNF; transgene; recombinant; diabetes; beta-cell; cDNA; beta-cell; 〓DNA; transgenic mouse; TNF-α; shock

I.We analyzed the structure of the 5' end of the mouse CD14 (mCD14) gene. Sequentially 5'-deleted, chimeric and point mutated clones were tested for their ability to stimulate chroramphnicol acetyltransferase (CAT). An 8-base pair sequence, TGATTCAC, at position -255 enhanced the expression of the CAT gene in macrophage cell lines. The 5' upstream of the human CD14 gene were sequenced and compared with that of mouse. II.Recombinant mCD14 protein was deleted at carboxyterminus was produced in E.coli. We made monoclonal antibodies (mAb) against the protein. Immunohistochemical staining using mAb showed that CD14-positive Kupffer cells (KC) were small in number in the liver in nonstimulated mice. The number of stained KC gradually increased with time after intraperitoneal injection of LPS, peaked 6 h after injection, and returned to normal by 20 h after injection. Slight increase in mCD14 expression was observed in peritoneal macrophages 2 h after LPS administration in vivo using flow cytometric analysis. Unique features of mCD14 expression in KC were also confirmed by the expression of mCD14 mRNA which peaked at 3 h after LPS administration. Mice bearing a mCD14 transgene controlled by an metallotionein promoter (MTTG) were produced. Northen blot analysis of MTTG showed that the mice strongly expressed mCD14 mRNA in the liver, small intestine and testis. III.Mice bearing a TNFalpha transgene controlled by an insulin promoter developed an increasingly severe insulitis. Diabetes never divelopped, even though a number of potentially inducing conditions were used, including the prolonged perfusion of TFNgamma and the permanent expression of a non-tolerogenic viral protein on beta cells.

项目成果

山本俊輔: "好中球、" 中山書店(印刷中), (1994)

山本俊辅：《中性粒细胞》，中山书店（正在出版），（1994）

Yasunori Higuchi: "Expression of a tumor necrosis factor-α transgene in murine pancreatic β cells results in severe and parmanent insulitis without evolution towards diabetes" J.Exp.Med.176. 1719-1731 (1992)

Yasunori Higuchi：“小鼠胰腺 β 细胞中肿瘤坏死因子-α 转基因的表达会导致严重且永久性的胰岛炎，但不会发展为糖尿病”J.Exp.Med.176-1719-1731 (1992)。

Matsuura, K.et al.: "Identification of a tissue-specific regulatory element in the murine CD14 gene" Journal Experimental Medicine. (in press).

Matsuura, K.et al.：“小鼠 CD14 基因中组织特异性调节元件的鉴定”实验医学杂志。

Ohashi, P.S.et al.: "Induction of diabetes is influenced by the infectious virus and local expression of MHC class I and tumor necrosis factor-alpha" Journal of Immunology. 150(11). 5185-5194 (1992)

Ohashi, P.S.等人：“糖尿病的诱发受到传染性病毒以及 MHC I 类和肿瘤坏死因子-α 局部表达的影响”《免疫学杂志》。

Higuchi, Y.et al.: "Expression of a tumor necrosis factor-alpha transgene in murine pancreatic beta cells results in severe and permanent insulitis without evolution towards diabetes" Journal Experimental Medicine. 176(6). 1719-1731 (1992)

Higuchi, Y.等人：“在小鼠胰腺β细胞中表达肿瘤坏死因子-α转基因会导致严重且永久性的胰岛炎，而不会演变成糖尿病”《实验医学杂志》。