A new hematopoietic cell line, KMT-2, having human interleukin-3 receptors

S. Tamura, M. Sugawara, H. Tanaka, E. Tezuka, S. Nihira, C. Miyamoto, T. Suda, Y. Ohta

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Abstract

A novel human cell line, KMT-2, from umbilical cord blood cells was established based on the selection of cultures in the presence of recombinant human interleukin-3 (IL-3) and the sorting of cells with anti-My 10 antibody. Morphologic and cytochemical studies (peroxidase negative, Sudan-black negative, chloroacetate esterase negative, PAS positive, nonspecific esterase positive) and phenotyping (HLA-DR, My7 = CD13, My9 = CD33, My10 = CD34, MCS-2, LeuM1 positive, glycophorin A negative, and P2 negative) suggest that the KMT-2 cells are myelomonocytic cells, probably of immature progenitor origin. Besides IL-3, granulocyte-macrophage colony-stimulating factor supported the growth of the KMT-2 cells, but IL-1α, IL-2, IL-4, IL-5, and erythropoietin did not. IL-6 showed only slight activity. Binding studies with 125I-labeled recombinant human (rh) IL-3 indicated that IL-3 bound to a single class of high affinity receptors (~ 4,000 receptors/cell) on KMT-2 cells with a kd of approximately 200 pmol/L. The chemical cross-linking assay demonstrated that radiolabeled hIL-3 bound three molecules with molecular masses of 170, 130, and 70 Kd. Present data suggest that the newly established human cell line will be a valuable tool for the biologic assay of hIL-3, and a model for biochemical studies of IL-3 receptors.

Original languageEnglish
Pages (from-to)501-507
Number of pages7
JournalBlood
Volume76
Issue number3
Publication statusPublished - 1990 Jan 1

ASJC Scopus subject areas

  • Biochemistry
  • Immunology
  • Hematology
  • Cell Biology

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    Tamura, S., Sugawara, M., Tanaka, H., Tezuka, E., Nihira, S., Miyamoto, C., Suda, T., & Ohta, Y. (1990). A new hematopoietic cell line, KMT-2, having human interleukin-3 receptors. Blood, 76(3), 501-507.