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

在我的研究期间，获得了以下结果：1)BALB/c和AKR/J系小鼠胸腺基质细胞系的建立；2)BALB/c系基质细胞系表达Thy 1分子。3)一些基质细胞系表达Src家族蛋白酪氨酸激酶，如Src和Fgr。用本实验室研制的抗FGR单抗2H2免疫组织化学检测，骨髓基质细胞呈FGR蛋白阳性。(微生物，免疫学，40：223,1996)4)当抗原肽与胸腺皮质和髓质上皮细胞系一起呈现时，DB14 T细胞克隆的效应功能被选择性地激活(imid.41：51,1997)。5)建立了15株针对AKR/J来源的基质细胞的单抗。用有限稀释法建立了4个无性系。单抗、HI-C、HI-M、HI-E和HI-F的组织和应变分布均为…进一步分析。6)将基质细胞三维培养于凝胶或高孔微珠上，进行体内移植。在被测试的材料中，基于明胶的多孔珠(Cultispher^&lt；TM&gt；，Percell Biolytica)和纤维蛋白凝块与明胶珠相结合，可能是转移或移植重组基质细胞或人工胸腺的生物相容性载体。7)移植的基质细胞在肾被膜下凝胶中可见一至多层基质细胞，并伴有单个核细胞的浸润。8)具有特定来源的间质细胞的胸腺组织重组应该在不丢失某些类型的间质细胞的情况下进行，因为在Aly/Aly(淋巴发育不全)突变小鼠的胸腺中，负选择似乎受到干扰，因为由于某些细胞类型或特定细胞上的某个分子的丢失而导致骨髓功能缺陷(Proc.Natl.Academy.USA 94：2472,1997)。较少

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