Artificial thymus-a completely re-organized thymic tissue with stromal cell lines of defined origins

人工胸腺——完全重组的胸腺组织，具有确定来源的基质细胞系

• 批准号: 07670360
• 负责人: IWABUCHI Kazuya
• 金额: $ 1.47万
• 依托单位: HOKKAIDO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07670360/
• 关键词: thymus; artificial thymus; stromal cells; cell transplantation; monoclonal antibody

During my term of project, following results were obtained.1) Thymic stromal cell lines were established from BALB/c and AKR/J strains of mice.2) One of stromal cell lines from BALB/c strain expressed Thy 1 molecules. An inhibitory signal for growth was shown to be transduced from Thy 1 molecules.3) Some of stromal cell lines were shown to express Src-family protein tyrosine kinases, such as Src and Fgr. And medullary stromal cells were positive for Fgr proteins by immunohistology with anti-Fgr mAb, 2H2 which was established in our laboratory. (Microbiol.Immunol.40 : 223,1996.)4) Selective activation for effector functions of DB14 T-cell clone was exhibited when an antigenic peptide was presented with thymic cortical and medullary epithelial cell lines (ibid.41 : 51,1997).5) Fifteen mAbs were established against stromal cells of AKR/J origin. Four clones of hydridoma were established with limiting dilution method. With mAbs, HI-C,HI-M,HI-E,and HI-F,tissue and strain distributions were … More analyzed.6) Stromal cells were cultured in 3-dimension in gel or on highly porous beads for in vivo transfer. Among materials tested, gelatin-based porous beads (Cultispher^<TM>, Percell Biolytica) and fibrin clot in combination with gelatin beads could be candidates for biocompatible carriers for transfer or transplantation of re-organized stromal cells, or artificial thymus. Biocompatibility, especially on adhesion between stromal cells and bead surface should be improved.7) Transplanted stromal cells in gel under kidney capsule were observed as one to several layrs of stromal cells with an infiltration of mononuclear cells. These artificial thymi in situ did not seem to function as thymic tissues but induced inflammatory responses.8) Re-organization of thymic tissue with stromal cells of defined origins should be performed without any loss of certain types of stromal cells, since negative selection seemed perturbed in the thymus of aly/aly (alymphoplasia) mutant mouse that has a defect in medullary function due to a loss of a certain cell type or a molecule on certain cell (Proc.Natl.Acad.Sci.USA 94 : 2472,1997). Less

在我的项目期间，获得了以下结果：1）从BALB/c和AKR/J品系小鼠中建立胸腺基质细胞系。2）BALB/c品系的基质细胞系之一表达Thy 1分子。生长的抑制信号被证明是从 Thy 1 分子转导的。3) 一些基质细胞系被证明表达 Src 家族蛋白酪氨酸激酶，例如 Src 和 Fgr。用本实验室建立的抗Fgr单克隆抗体2H2进行免疫组化检测，骨髓基质细胞Fgr蛋白呈阳性。 (Microbiol.Immunol.40：223，1996。)4)当胸腺皮质和髓质上皮细胞系呈递抗原肽时，表现出对 DB14 T 细胞克隆的效应器功能的选择性激活(同上。41：51，1997)。5)针对 AKR/J 来源的基质细胞建立了 15 种 mAb。采用有限稀释法建立了4个杂交瘤克隆。使用 mAb、HI-C、HI-M、HI-E 和 HI-F，对组织和菌株分布进行了分析。6) 将基质细胞在凝胶中或在高度多孔的珠子上进行 3 维培养，以进行体内转移。在测试的材料中，基于明胶的多孔珠(Cultispher TM ，Percell Biolytica)和与明胶珠组合的纤维蛋白凝块可以是用于转移或移植重组基质细胞或人工胸腺的生物相容性载体的候选者。生物相容性，特别是基质细胞与珠表面之间的粘附力有待提高。7)肾被膜下凝胶中移植的基质细胞观察到一层至几层基质细胞，并有单核细胞浸润。这些原位人造胸腺似乎并不具有胸腺组织的功能，而是诱导炎症反应。8）应在不损失某些类型基质细胞的情况下用确定来源的基质细胞重组胸腺组织，因为 aly/aly（发育不全）突变小鼠的胸腺中的负选择似乎受到干扰，该小鼠因丢失某种细胞而导致髓质功能缺陷 某些细胞上的类型或分子(Proc.Natl.Acad.Sci.USA 94：2472，1997)。较少的

项目成果

Kaneda Rika: "Detachment activity but not cytotoxicity induced in a T-cell clone following antigen presentation in the presence of thymic epithelial cells." Microbiol.Immunol.41 (1). 51-61 (1997)

Kaneda Rika：“在胸腺上皮细胞存在的情况下，抗原呈递后，T 细胞克隆会诱导脱离活性，但不会诱导细胞毒性。”

Kaneda Rika: "Dissociation of Fas-mediated cytotoxicity and FasL expression in a cytotoxic CD4^+ T cell clone." Immunol.Lett.55. 53-60 (1997)

Kaneda Rika：“细胞毒性 CD4^ T 细胞克隆中 Fas 介导的细胞毒性和 FasL 表达的分离。”

Nakagawa Ken-ichi: "Generation of NK1.1+TCRα/β^+ thymocytes associated with intact thymic structure." Proc. Natl. Acad. Sci. USA. 94(in press). (1997)

Ken-ichi Nakakawa：“与完整胸腺结构相关的 NK1.1+TCRα/β^+ 胸腺细胞”，美国国家科学院院刊 94。

Itoh Yasushi: "Determination of the allele-specific antigen-binding site on I-A^k and I-A^b molecules." Eur. J. Immunol.26. 1314-1321 (1996)

Itoh Yasushi：“确定 I-A^k 和 I-A^b 分子上的等位基因特异性抗原结合位点。”

Kaneda Rika: "Dissociation of Fas-mediated cytotoxicity and FasL expression in a cytotoxic CD4+T cell clone." Immunol. Lett.(in press). (1997)

Kaneda Rika：“细胞毒性 CD4 T 细胞克隆中 Fas 介导的细胞毒性和 FasL 表达的分离。”