Turner A M, Bennett L G, Lin N L, Wypych J, Bartley T D, Hunt R W, Atkins H L, Langley K E, Parker V, Martin F
Division of Hematology, University of Washington, Seattle 98195, USA.
Blood. 1995 Apr 15;85(8):2052-8.
Stem cell factor (SCF) triggers cell growth by binding to cell surface c-kit receptors. Soluble forms of several cytokine receptors have been described and may play a role in the modulation of cytokine activity in vivo. For these reasons, we investigated whether human hematopoietic cells produce soluble c-kit receptors. The human leukemia cell lines OCIM1 and MO7e display approximately 80,000 and approximately 35,000 high-affinity cell surface c-kit receptors, respectively. Soluble c-kit receptors were detected by enzyme immunoassay in OCIM1 and MO7e culture supernatants. We determined the molecular weight and binding affinity of soluble c-kit receptor produced by OCIM1 cells, soluble c-kit receptor purified from human serum, and recombinant soluble c-kit receptor expressed in CHO cells. The three soluble c-kit receptors each have a molecular weight of 98 kD. Quantitative binding experiments with 125I-SCF indicate that the soluble c-kit receptors obtained from human serum or OCIM1 cells have binding affinities for SCF of approximately 200 to 300 pmol/L, in contrast to the recombinant form, which has a binding affinity of approximately 1.5 nmol/L. All three forms of the soluble c-kit receptor were able to compete with c-kit receptors on OCIM1 cells for 125I-SCF binding. Thus human hematopoietic cells can produce a soluble form of the c-kit receptor that retains high-affinity SCF binding activity. We speculate that the soluble c-kit receptor may bind SCF and function as a receptor antagonist in vivo.
干细胞因子(SCF)通过与细胞表面的c-kit受体结合来触发细胞生长。已经描述了几种细胞因子受体的可溶性形式,它们可能在体内细胞因子活性的调节中发挥作用。基于这些原因,我们研究了人类造血细胞是否产生可溶性c-kit受体。人类白血病细胞系OCIM1和MO7e分别显示约80,000和约35,000个高亲和力细胞表面c-kit受体。通过酶免疫测定法在OCIM1和MO7e培养上清液中检测到可溶性c-kit受体。我们测定了OCIM1细胞产生的可溶性c-kit受体、从人血清中纯化的可溶性c-kit受体以及在CHO细胞中表达的重组可溶性c-kit受体的分子量和结合亲和力。这三种可溶性c-kit受体的分子量均为98 kD。用125I-SCF进行的定量结合实验表明,从人血清或OCIM1细胞获得的可溶性c-kit受体对SCF的结合亲和力约为200至300 pmol/L,而重组形式的结合亲和力约为1.5 nmol/L。可溶性c-kit受体的所有三种形式都能够与OCIM1细胞上的c-kit受体竞争125I-SCF的结合。因此,人类造血细胞可以产生一种可溶性形式的c-kit受体,该受体保留高亲和力SCF结合活性。我们推测可溶性c-kit受体可能在体内结合SCF并作为受体拮抗剂发挥作用。