A new hematopoietic cell line, KMT-2, having human interleukin-3 receptors.
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-1 alpha, 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 (approximately 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.
Department of Oncology, Nippon Roche Research Center, Kamakura City, Japan.
S
S
Tamura
M
M
Sugawara
H
H
Tanaka
E
E
Tezuka
S
S
Nihira
C
C
Miyamoto
T
T
Suda
Y
Y
Ohta
D001773:Blood Cells / Q000378:metabolism / Q000648:ultrastructure*; D002460:Cell Line; D003115:Colony-Stimulating Factors / Q000494:pharmacology; D005312:Fetal Blood / Q000166:cytology; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D006133:Growth Substances / Q000494:pharmacology; D006801:Humans; D007377:Interleukin-3 / Q000378:metabolism / Q000502:physiology; D011971:Receptors, Immunologic / Q000378:metabolism* / Q000502:physiology; D016185:Receptors, Interleukin-3
D003115:Colony-Stimulating Factors; D006133:Growth Substances; D007377:Interleukin-3; D011971:Receptors, Immunologic; D016185:Receptors, Interleukin-3; D016178:Granulocyte-Macrophage Colony-Stimulating Factor