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

在我的项目期间，取得了以下成果：1)利用小鼠BALB/c和AKR/J株建立小鼠胸腺基质细胞系。2) BALB/c株间质细胞株中有1株表达th1分子。一个抑制生长的信号被证明是由th1分子转导的。3)部分间质细胞系表达Src家族蛋白酪氨酸激酶，如Src和Fgr。骨髓间质细胞用本实验室建立的抗Fgr单抗2H2免疫组织学检测Fgr蛋白阳性。(Microbiol.Immunol。[40][23] 1996。4)在胸腺皮质和髓质上皮细胞系中添加抗原肽可选择性激活DB14 t细胞克隆的效应功能（同上41:51,1997）。5)建立了15个针对AKR/J来源基质细胞的单克隆抗体。用极限稀释法建立了4个水螅无性系。使用单克隆抗体，HI-C,HI-M，HI-E和HI-F，组织和菌株分布分析。6)基质细胞在凝胶或高孔珠上三维培养，用于体内转移。在测试的材料中，明胶基多孔微球（Cultispher^<TM>, Percell Biolytica）和纤维蛋白凝块与明胶微球的结合可以作为生物相容性载体，用于转移或移植重组基质细胞或人工胸腺。生物相容性，特别是基质细胞与珠表面的粘附性有待提高。7)肾被膜下移植的凝胶间质细胞为一至多层间质细胞，并有单核细胞浸润。这些人工胸腺原位似乎没有胸腺组织的功能，但诱导炎症反应。8)胸腺组织与起源明确的基质细胞的重组应该在不丢失任何特定类型基质细胞的情况下进行，因为在aly/aly（淋巴增生）突变小鼠的胸腺中，由于某种细胞类型或某种细胞上的分子的丢失而导致髓质功能缺陷，因此负选择似乎受到了干扰（procc . natl . acad . sci）。美国94:2472,1997)。少

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