Generation of artificial lymphoid tissues and their application.

结果表明，人工淋巴组织不仅具有正常淋巴组织的基本结构，而且具有产生抗原特异性抗体的重要免疫功能。为了构建“人工淋巴组织”结构，制备了一种含有胸腺源性基质细胞系和细胞因子的生物相容性材料，并将其移植到小鼠肾包膜下间隙。在移植时，骨髓来源的成熟树突状细胞（DCs）与基质细胞一起加入移植物中，加速了淋巴细胞的积累。许多较大的T细胞灶周围有B细胞聚集，类似于继发性淋巴组织中的T细胞区和B细胞区。进一步的免疫组化分析显示，大量T细胞和B细胞积累的移植物在以下几个方面的结构与次级淋巴组织相似；1) CD11c +树突细胞被发现散落在T细胞和B细胞移植积累,2)集群中的CD4 / CD8 T细胞的比例是类似于T细胞地区正常淋巴结组织,3)的B细胞集群包含CR1积极树突细胞中心,表明滤泡树突形成网络,4)花生凝集素(机构)积极的B细胞被发现在一些B细胞集群,5)Syndecan-1积极的浆细胞被发现在B细胞积累,6)PNAd（外周淋巴结寻址蛋白）或MAdCAM-1（粘膜寻址蛋白细胞粘附分子-1）阳性HEV（高内皮小静脉）样结构在部分移植物中（特别是）发现大量T细胞和B细胞积聚。此外，通过酶联免疫斑点（ELISPOT）试验和免疫组织化学分析证实移植物中抗原特异性抗体的形成。

We showed that our artificial lymphoid tissues possess not only basic structure of normal lymphoid tissue, but also immunologically important function of antigen specific antibody production. To construct "artificial lymphoid tissue" structures, a certain biocompatible material containing a thymus derived stromal cell line and cytokines was prepared, and transplanted to renal subcapsular space in mice. The accumulation of the lymphocytes was accelerated by the addition of bone marrow derived mature dendritic cells(DCs) in the graft along with the stromal cells at the time of transplantation.Many and relatively large T cell foci surrounded by B cell accumulation, which is similar to T cell and B cell area in secondary lymphoid tissues. Further immunohistochemical analysis revealed that the grafts with many T cell and B cell accumulation structurally resembled secondary lymphoid tissues in following aspects(points) ; 1)CD11c+ dendritic cells were found scattered both in T cell and B cell accumulation in the graft, 2)the CD4/CD8 ratio of T cells in the cluster was similar to that of T cell areas in normal lymph node tissues, 3)some of B cell clusters contained CR1 positive dendritic cells in the center, suggesting follicular dendritic network formation, 4)peanut agglutinin(PNA) positive B cells were detected in some of B cell clusters, 5)Syndecan-1 positive plasma cells were found around B cell accumulation, and 6)PNAd(peripheral node addressin) or MAdCAM-1(mucosal addressin cell adhesion molecule-1) positive HEV(high endothelial venule)-like structure was found in some of the grafts(especially) with many T cell and B cell accumulation.In addition, antigen specific antibody formation in the grafts was confirmed by enzyme-linked immunospot(ELISPOT) assay and immunohistochemical analysis.