Garcia-Pardo A, Ferreira O C
New York Blood Center, NY 10021.
Immunology. 1990 Jan;69(1):121-6.
Lymphocyte adhesion to components of extracellular matrices (i.e. fibronectin) is important for their proper localization in tissues and inflammatory sites. We have studied the attachment of the human cell line HUT-78 (mature T lymphocytes) to fibronectin and to several tryptic fragments of fibronectin. HUT-78 cells effectively adhered to surfaces coated with two Hep II domain-containing fragments of 38,000 and 58,000 MW derived from the A and B chains of fibronectin, respectively. Cells also bound to an 80,000 MW fragment containing the RGDS sequence of fibronectin. Cell adhesion to the 38,000 MW fragment was completely inhibited (100%) by cell preincubation with the soluble 38,000 MW fragment; it was partially inhibited (30-37%) by preincubation with the 58,000 MW fragment or with a synthetic peptide CS-1, comprising the first 25 amino acid residues of the alternatively spliced connecting segment (IIICS), which is present in the A chain of fibronectin and in the 38,000 MW fragment. Cell preincubation with RGDS-containing synthetic peptides or with the 80,000 MW fragment, did not affect attachment to 38,000 MW-coated surfaces. Moreover, preincubation of HUT-78 cells with 38,000 MW fragment had no effect on cell adhesion to 80,000 MW-coated wells, while preincubation with 80,000 MW fragment completely inhibited cell attachment to these surfaces. These results strongly suggest the involvement of two different cell surface receptors which recognize the Hep II/IIICS site and the RGDS site independently. Preincubation with either 38,000 or 80,000 MW fragments prevented cell attachment to fibronectin, indicating that adhesion to the intact molecule requires interaction with both regions. Therefore T-lymphocyte adherence to fibronectin-containing matrices may be regulated by the co-expression of both receptors at the cell surface.
淋巴细胞与细胞外基质成分(如纤连蛋白)的黏附对于它们在组织和炎症部位的正确定位很重要。我们研究了人细胞系HUT-78(成熟T淋巴细胞)与纤连蛋白以及纤连蛋白的几个胰蛋白酶片段的附着情况。HUT-78细胞有效地黏附于分别由纤连蛋白A链和B链衍生的两个含Hep II结构域、分子量为38,000和58,000的片段包被的表面。细胞也与一个含有纤连蛋白RGDS序列的80,000分子量片段结合。细胞与38,000分子量片段的黏附在与可溶性38,000分子量片段预孵育后被完全抑制(100%);与58,000分子量片段或与一个合成肽CS-1(包含可变剪接连接段(IIICS)的前25个氨基酸残基,该连接段存在于纤连蛋白A链和38,000分子量片段中)预孵育后被部分抑制(30 - 37%)。用含RGDS的合成肽或80,000分子量片段对细胞进行预孵育,不影响其对38,000分子量包被表面的附着。此外,用38,000分子量片段对HUT-78细胞进行预孵育对其与80,000分子量包被孔的黏附没有影响,而用80,000分子量片段预孵育则完全抑制细胞对这些表面的附着。这些结果有力地表明涉及两种不同的细胞表面受体,它们分别独立识别Hep II/IIICS位点和RGDS位点。用38,000或80,000分子量片段进行预孵育均可阻止细胞与纤连蛋白的附着,这表明与完整分子的黏附需要与两个区域相互作用。因此,T淋巴细胞与含纤连蛋白基质的黏附可能受细胞表面两种受体共表达的调节。