Stoolman L M, Wang T L, Situ R, Varani J
Department of Pathology, University of Michigan Medical School, Ann Arbor 48109-0602.
J Cell Physiol. 1993 Mar;154(3):593-600. doi: 10.1002/jcp.1041540318.
The current study shows that a clonal derivative of the Jurkat cell line up-regulates both the avidity and density of the alpha 6/beta 1 receptor in response to phorbol 12-myristate 13-acetate (PMA). This derivative attaches to fibronectin and, to a lesser degree, laminin constitutively. Adhesion and spreading are dramatically up-regulated following treatment with PMA. The response on fibronectin peaks within 4 hours, is insensitive to cyclohexamide, can be blocked by monoclonal antibodies (Mabs) to the beta 1 and alpha 5 subunits of the beta 1 family of integrins, and is not associated with increased expression of the alpha 5 or beta 1 epitopes at the cell surface. In contrast, the response on laminin is biphasic. The early phase parallels the response on fibronectin. The second phase peaks after 48-72 hours of treatment with PMA, is sensitive to cycloheximide, can be blocked by Mabs to the beta 1 and alpha 6 subunits, and is associated with increased expression of the alpha 6 epitope. Both the density independent and dependent responses to PMA in Jurkat cells are blocked by the protein kinase inhibitor staurosporine. The HSB-2, CEM, Molt-4, and HPB-ALL T-lymphoblastic cell lines also up-regulate attachment to fibronectin and laminin following treatment with PMA. All four lines constitutively attach to fibronectin and show rapid up-regulation of attachment following treatment with PMA. None of the lines attach to laminin prior to PMA treatment; however, specific adhesion developed after 4-120 hours of treatment. The most mature lines (Jurkat and HPB-ALL) up-regulated adhesion on laminin more rapidly than the less phenotypically mature lines (CEM, Molt-4, and HSB-2). In summary, clonal derivatives of the Jurkat cell line up-regulated attachment to laminin through protein kinase dependent increases in alpha 6/beta 1 receptor avidity and density. In addition, the expression of functional receptors for laminin is linked to developmental maturity in a series of T-lymphoblastic cell lines.
当前研究表明,Jurkat细胞系的一个克隆衍生物在佛波醇12 - 肉豆蔻酸酯13 - 乙酸酯(PMA)作用下,上调α6/β1受体的亲和力和密度。该衍生物组成性地附着于纤连蛋白,对层粘连蛋白的附着程度较低。用PMA处理后,黏附与铺展显著上调。对纤连蛋白的反应在4小时内达到峰值,对环己酰亚胺不敏感,可被针对整合素β1家族β1和α5亚基的单克隆抗体(Mab)阻断,且与细胞表面α5或β1表位的表达增加无关。相比之下,对层粘连蛋白的反应是双相的。早期阶段与对纤连蛋白的反应相似。第二阶段在PMA处理48 - 72小时后达到峰值,对环己酰亚胺敏感,可被针对β1和α6亚基的Mab阻断,且与α6表位的表达增加有关。Jurkat细胞中对PMA的密度非依赖性和依赖性反应均被蛋白激酶抑制剂星形孢菌素阻断。HSB - 2、CEM、Molt - 4和HPB - ALL T淋巴细胞系在用PMA处理后也上调对纤连蛋白和层粘连蛋白的附着。所有四个细胞系都组成性地附着于纤连蛋白,并用PMA处理后显示出附着的快速上调。在PMA处理前,没有一个细胞系附着于层粘连蛋白;然而,在处理4 - 120小时后出现特异性黏附。最成熟的细胞系(Jurkat和HPB - ALL)比表型较不成熟的细胞系(CEM、Molt - 4和HSB - 2)更快地上调对层粘连蛋白的黏附。总之,Jurkat细胞系的克隆衍生物通过蛋白激酶依赖性增加α6/β1受体的亲和力和密度,上调对层粘连蛋白的附着。此外,在一系列T淋巴细胞系中,层粘连蛋白功能性受体的表达与发育成熟度相关。