Spleen-derived stromal cells. Adhesion molecules expression and lymphocyte adhesion to reticular cells.
Spleen-derived stromal cells. Adhesion molecules expression and lymphocyte adhesion to reticular cells.
复制标题
脾源性基质细胞。
DOI：
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发表时间：
1997
影响因子：
6.6
通讯作者：
M. Rosemblatt
中科院分区：
文献类型：
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作者：
A. Castro;M. Bono;V. Simón;L. Vargas;M. Rosemblatt
淋巴细胞分化的最后步骤发生在次级淋巴器官中，其中B和T淋巴细胞与淋巴微环境相互作用。虽然许多研究描述了小鼠淋巴细胞与树突状细胞、滤泡细胞和其他抗原呈递细胞的相互作用，但对淋巴细胞与脾基质的重要细胞组分网状细胞之间的相互作用知之甚少。在这项工作中，我们描述了培养完整的小鼠脾基质和两个细胞系，SP-1和SP-2，确定为可能的网状起源，并描述了小鼠淋巴细胞和人淋巴细胞与小鼠脾基质细胞之间的粘附相互作用。流式细胞仪分析表明，Sp-1细胞系显示出组成型表达VCAM-1和CD 44的单一细胞类型。它们不表达滤泡细胞、交错细胞、巨噬细胞或内皮细胞所述的任何标志物。我们的数据表明，这些细胞代表了脾网状细胞的群体。Sp-2细胞系显示两种表型不同的细胞类型，它们联合生长。流式细胞仪分析表明，这两种细胞类型表达VCAM-1和CD 44组成，但它们可以通过表达CD 11b和FcR的分化。这些数据表明，SP-2细胞系是由一种类型的基质细胞生长在网状细胞的粘附层。此外，对从鼠脾制备的非B非T细胞部分的分析显示，这些细胞中约30%对应于CD 44/VCAM-1双阳性细胞。鼠B和T细胞粘附于完整的基质和Sp-1和Sp-2细胞系。用LPS激活B细胞对结合没有影响，而Con-A处理后T细胞与完整基质的结合增加了3倍。人淋巴母细胞样Daudi细胞与完整脾基质的粘附被抗（鼠）VCAM-1抗体阻断，但不被（人）整联蛋白α 4亚基的抗体阻断，而与Sp-1和Sp-2基质的粘附被抗两种分子的抗体阻断。此外，拉莫斯细胞与Sp-2基质的粘附被整合素α 4亚基和鼠VCAM-1的抗体抑制。其他粘附受体如整合素β 2亚单位、ICAM-1或CD 44的抗体对人细胞与这些基质的结合没有影响。我们的研究结果表明，我们已经分离出一部分脾网状细胞，这些细胞可以培养为一个独特的细胞系。这些细胞组成性表达CD 44和VCAM-1，并使用这些分子中的一些用于淋巴细胞结合，这一发现表明脾网状细胞可能参与调节通过脾的正常淋巴细胞运输。
The final steps of lymphocyte differentiation occur in secondary lymphoid organs where B and T lymphocytes interact with the lymphoid microenvironment. Although numerous studies describe the interactions of murine lymphocytes with dendritic, follicular and other antigen presenting cells, little is known on the interactions between lymphocytes and reticular cells, an important cellular component of spleen stroma. In this work we describe the culturing of complete murine spleen stromas and of two cell lines, Sp-1 and Sp-2, identified as of possible reticular origin, and describe the adhesive interactions between murine lymphocytes and human lymphoid cells with murine spleen stromal cells. FACS analysis indicates that the Sp-1 cell line shows a single cell type expressing VCAM-1 and CD44 constitutively. They do not express any of the markers described for follicular cells, interdigitating cells, macrophages or endothelial cells. Our data suggests that these cells represent a population of spleen reticular cells. The Sp-2 cell line shows two phenotypically different cell types that grow in association. FACS analysis demonstrates that both cell types express VCAM-1 and CD44 constitutively, but that they can be differentiated by the expression of CD11b and FcR. These data suggest that the Sp-2 cell line is composed of one type of stromal cell growing over an adherent layer of reticular cells. Furthermore, analysis of the non-B non-T cell fraction prepared from murine spleen shows that approximately 30% of these cells correspond to the CD44/VCAM-1 double positive cells. Murine B and T cells adhere to the complete stromas and to Sp-1 and Sp-2 cell lines. Activation of B cells with LPS had no effect on binding while binding of T cells to complete stromas increased up to threefold after Con-A treatment. Adhesion of human lymphoblastoid Daudi cells to complete spleen stromas is blocked by an anti-(murine) VCAM-1 antibody but not by an antibody to the (human) integrin alpha 4 subunit, while adhesion to the Sp-1 and Sp-2 stromas is blocked by antibodies against both molecules. Also, adhesion of Ramos cells to Sp-2 stromas is inhibited by antibodies to the integrin alpha 4 subunit and to murine VCAM-1. Antibodies to other adhesion receptors such as the integrin beta 2 subunit, ICAM-1 or CD44 have no effect on human cell binding to these stromas. Our results suggest that we have isolated a fraction of splenic reticular cells and that these cells can be cultured as a distinct cell line. The finding that these cells express CD44 and VCAM-1 constitutively and use some of these molecules for lymphocyte binding suggests that spleen reticular cells may be involved in the regulation of normal lymphocyte traffic through the spleen.