Establishment and characterization of a novel human monocytic leukemia cell line (KP-1) expressing scavenger receptor.
A new monocytic leukemia cell line (KP-1) was established from a 2-y-old Japanese girl with acute monocytic leukemia. The KP-1 cells were maintained in suspension culture with a doubling time of 96 h. The cells were positively stained with alpha-naphtyl butyrate esterase, but not with naphthol AS-D chloroacetate esterase, myeloperoxidase, and periodic acid-Schiff reagent. Cell surface marker analysis revealed that the cells were CD4, CD11a, CD11c, CD13, CD14, CD18, CD33, and HLA-DR positive. Karyotype analysis revealed near diploidy (47 XX) and a translocation t(11;19) was found. When treated with 12-o-tetradecanoylphorbol 13-acetate, KP-1 cells became tightly adherent, showed the enhanced reactivity for alpha-naphtyl butyrate esterase, and produced several monokines such as IL-1 beta, tumor necrosis factor-alpha, and macrophage colony-stimulating factor. Immunoelectron microscopy demonstrated that the human macrophage scavenger receptor was expressed after 12-o-tetradecanoylphorbol-13-acetate treatment, and the cells accumulated a large amount of cholesterol esters in the presence of acetylated LDL. Compared with another human monocytic leukemia cell line, THP-1, KP-1 expressed scavenger receptor and accumulated cholesterol ester more rapidly in the presence of 12-o-tetradecanoyl phorbol-13-acetate and acetylated LDL. Scatchard analysis using 125I-labeled acetylated LDL revealed a typical saturation curve with an apparent kd of 1.7 x 10(-7) M and 3400 binding sites per cell. KP-1 retained the characteristics of monocyte-macrophage lineage cells and will facilitate the in vitro studies of the pathologic and physiologic roles of scavenger receptors.
Adachi
N
N
Department of Pediatrics, Kumamoto University School of Medicine, Japan.
Takaki
K
K
Nakamura
R
R
Ohta
T
T
Naito
M
M
Horiuchi
S
S
Matsuda
I
I
D000951:Antigens, Neoplasm / Q000032:analysis; D014408:Biomarkers, Tumor / Q000032:analysis; D002454:Cell Differentiation / Q000187:drug effects; D002675:Child, Preschool; D002788:Cholesterol Esters / Q000378:metabolism; D002869:Chromosome Aberrations; D005260:Female; D015973:Gene Expression Regulation, Leukemic / Q000187:drug effects; D006801:Humans; D007621:Karyotyping; D007948:Leukemia, Monocytic, Acute / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D008077:Lipoproteins, LDL / Q000494:pharmacology; D008264:Macrophages / Q000378:metabolism / Q000473:pathology; D008565:Membrane Proteins; D016253:Microscopy, Immunoelectron; D015846:Monokines / Q000096:biosynthesis; D009363:Neoplasm Proteins / Q000096:biosynthesis*; D014411:Neoplastic Stem Cells / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D010587:Phagocytosis; D011971:Receptors, Immunologic / Q000096:biosynthesis*; D018110:Receptors, Lipoprotein; D051116:Receptors, Scavenger; D051122:Scavenger Receptors, Class B; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014407:Tumor Cells, Cultured / Q000378:metabolism
D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; D002788:Cholesterol Esters; D008077:Lipoproteins, LDL; D008565:Membrane Proteins; D015846:Monokines; D009363:Neoplasm Proteins; D011971:Receptors, Immunologic; D018110:Receptors, Lipoprotein; D051116:Receptors, Scavenger; C454739:Scarb1 protein, mouse; D051122:Scavenger Receptors, Class B; D013755:Tetradecanoylphorbol Acetate