Sugimoto K, Tsurumaki Y, Hoshi H, Kadowaki S, LeBousse-Kerdiles M C, Smadja-Joffe F, Mori K J
Department of Physiology and Biochemistry, Faculty of Science, Niigata University, Japan.
Exp Hematol. 1994 Jun;22(6):488-94.
Cocultivation of erythroid leukemic cells (ELM-I-1) with hemopoietic supportive cells (MS-5) resulted in a specific adhesion of ELM-I-1 cells to MS-5 cells. This phenomenon was designated as rosette formation. After induction of differentiation of ELM-I-1 cells, rosette formation was reduced, and no rosette formation was observed between erythrocytes and MS-5 cells. Studies on anti-adhesion molecule antibody treatment have revealed that CD44 plays a key role in rosette formation. Expression of CD44 on (the membrane of) ELM-I-1 cells was reduced after differentiation, and no CD44 expression was detected on erythrocytes. CD44 was also expressed on MS-5. Hyaluronate is known as the ligand of CD44, but neither hyaluronidase treatment nor addition of excess hyaluronate to the assay system affected rosette formation. These data indicate that hyaluronate is not responsible for rosette formation. Anti-CD44 antibody (KM81), which recognized the hyaluronate binding site of CD44, inhibited rosette formation. But other monoclonal antibodies against different epitopes except for the hyaluronate binding site, even those against CD44's hyaluronate binding site, did not inhibit rosette formation. Thus, rosette formation between MS-5 cells and ELM-I-1 cells is mediated by CD44 but not by the hyaluronate binding site of CD44.
红白血病细胞(ELM-I-1)与造血支持细胞(MS-5)共培养导致ELM-I-1细胞与MS-5细胞发生特异性黏附。这种现象被称为玫瑰花结形成。ELM-I-1细胞诱导分化后,玫瑰花结形成减少,并且在红细胞与MS-5细胞之间未观察到玫瑰花结形成。抗黏附分子抗体处理研究表明,CD44在玫瑰花结形成中起关键作用。分化后ELM-I-1细胞(膜)上CD44的表达降低,红细胞上未检测到CD44表达。MS-5上也表达CD44。透明质酸是已知的CD44配体,但透明质酸酶处理或向检测系统中添加过量透明质酸均不影响玫瑰花结形成。这些数据表明透明质酸与玫瑰花结形成无关。识别CD44透明质酸结合位点的抗CD44抗体(KM81)抑制玫瑰花结形成。但针对除透明质酸结合位点外不同表位的其他单克隆抗体,即使是针对CD44透明质酸结合位点的抗体,也不抑制玫瑰花结形成。因此,MS-5细胞与ELM-I-1细胞之间的玫瑰花结形成是由CD44介导的,而非由CD44的透明质酸结合位点介导。