Analysis of a fuctor involving trnslocation of LT to the cell.

涉及 LT 转移至细胞的功能因子的分析。

• 批准号: 09670302
• 负责人: TSUJI Takao
• 金额: $ 1.92万
• 依托单位: Fujita Health University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09670302/
• 关键词: ETEC; Entocytosis; GM1 ganglioside; Enterotoxin (LT); LT

We have been analyzing whether there might be a factor involving the endocytosis of ETEC enterotoxin (LT) to target cell except GMI ganglioside receptor. After LT was added to the CHO cell, Immunoprecipitates with anti-LT antibody to cell lysate was analyzed with Western blotting with anti-LT antibody. There was a band, of which molecular weight was 22,000 except the monomer of LT-B and might shift the monomer upper side of gel. Then we examined this factor. However, some other examiner reported that this shift of the monomer was caused by the formation of dimer of LT-B in the cell. Therefore, we examined whether other factor might be involved in the endocytosis of LT to make an antibody blocking the endocytosis of LT-B.Some rabbits were immunized by cell lysate, which was prepared from NCTC 2071 defecting GM1 ganglioside and we can prepared the anti-serum partially blocking the internization of LT to CHO cells. Then, we are now examining a factor reacting to this antiserum.Morecover, while we received this grant, we reported data as followed;1. LT is also coded in the chromosome, except the plasmid.2. The mutant defecting nicking site of A subunit near Arg 192 have strong adjuvant action to VZV, KKLH, Ovalbumin, BGG and measle virus.3. The nicking site near Arg 192 of the A subnit is not involved in some biological activity of LT.

我们一直在分析除GMI神经节受体以外的靶细胞的ETEC肠毒素（LT）的内吞作用是否可能存在因素。将LT添加到CHO细胞中后，用抗LT抗体的蛋白质印迹分析了用抗LT抗体的抗LT抗体的免疫沉淀物。有一个频带，其中分子量为22,000，除了LT-B的单体，可能会移动凝胶的单体上侧。然后我们检查了这个因素。但是，其他一些检查员报告说，单体的这种转移是由细胞中LT-B的二聚体形成引起的。因此，我们检查了是否可能参与LT的内吞作用，以使抗体阻断LT-B.LT-B.Some兔的内吞作用。细胞裂解液免疫，细胞裂解液是由NCTC 2071制备的，该抗体是由NCTC 2071降临的GM1 Ganglioside制备的，我们可以准备好抗serum的部分抗体，使抗seraum部分阻止了Lt to Cho to Cho Cho Chos to Cho Chos to Cho Chos的固化。然后，我们现在正在检查对这种抗血清反应的因素。当我们收到此赠款时，我们报告了遵循的数据； 1。 LT还在染色体中编码，除质粒2。 Arg 192附近亚基的突变体缺陷迹象对VZV，KKLH，Ovalbumin，BGG和Mesle病毒具有强大的辅助作用。3。 A子名称的ARG 192附近的昵称不参与LT的某些生物学活性。

项目成果

Tsuji T., Yokochi T., Kamiya H., Kawamoto K., Miyama A., and Asano Y.: "Relationship between a low toxicity of the mutant A subunit of enterotoxigenic Escherichia coli enterotoxin and its strong adjuvant action"Immunology. 90. 176-182 (1997)

Tsuji T.、Yokochi T.、Kamiya H.、Kawamoto K.、Miyama A. 和 Asano Y.：“产肠毒素大肠杆菌肠毒素突变 A 亚基的低毒性与其强佐剂作用之间的关系”免疫学。

Takao Tsuji et al: "Relationship between alow to xicity of the wutornt A sclt of entero toxigenic E colientenotoxin and its strony adjuvant action" Immurology. 90. 176-182 (1997)

Takao Tsuji 等人：“肠毒素 E colientenotoxin 的 wutornt A sclt 的低毒性与其强佐剂作用之间的关系”免疫学。

Tsuji T., Kato M., Kawase H., Imamura S., Kamiya H., Ichinose Y. and Miyama A.: "Escherichia coli enterotoxin demonstrates partial toxicity independent of the nicking around Arg192"Microbiology. 143. 1797-1804 (1997)

Tsuji T.、Kato M.、Kawase H.、Imamura S.、Kamiya H.、Ichinose Y. 和 Miyama A.：“大肠杆菌肠毒素表现出部分毒性，与 Arg192 周围的切口无关”微生物学。

Sasaki K., Kobayashi T., Imamura S., Shigekura T., Kado R., Kawamoto K., Tsuji T. and Miyama A.: "Flow cytometry analysis of the Fas ligand expression of activated lymph node T-cells"Immunol.Lett.. 55. 1-13 (1997)

Sasaki K.、Kobayashi T.、Imamura S.、Shigekura T.、Kado R.、Kawamoto K.、Tsuji T. 和 Miyama A.：“活化淋巴结 T 细胞 Fas 配体表达的流式细胞术分析”Immunol

Takao Tsuji et al: "E coli LT enterotoxin subunit A demoustvated partial toxicity in deroeudent of the nicking avoued A Ang192" Microhiology. 143. 1797-1804 (1997)

Takao Tsuji 等人：“大肠杆菌 LT 肠毒素亚基 A 在 deroeudent of the nicking avoued A Ang192 中消除了部分毒性”微生物学。