Phenotypic characterization of the human mast-cell line HMC-1.
The cell line HMC-1, derived from a patient with mast cell leukaemia, is the only established cell line exhibiting a phenotype similar to that of human mast cells. This paper reports on a detailed characterization of the expression of a panel of markers for various types of immature and mature haematopoietic cells in the HMC-1. We also studied the potential of HMC-1 to differentiate upon treatment with conditioned media from the human T-cell line Mo, retinoic acid or DMSO. HMC-1 was found to express several mast cell-related markers. A high expression of Kit, the receptor for stem-cell factor, was detected. The majority of the cells were stained with a MoAb against the mast cell-specific serine protease tryptase. Of particular interest was the finding that beta-tryptase mRNA, but not alpha-tryptase mRNA, was expressed in HMC-1. Using enzyme-histochemistry we were able to show that the beta-tryptase was enzymatically active, indicating that tryptase can form active homotetramers. Both heparin and chondroitin sulfate were found to be present in approximately equal amounts. HMC-1 lacked surface expression of the high-affinity IgE receptor, which was confirmed by the absence of mRNA of the alpha- and beta-chains of the IgE-receptor complex. However, a strong expression of the gamma-chain of the IgE-receptor complex was detected. A positive staining of the monocyte/macrophage marker CD68 was obtained, as well as a strong hybridization signal for the eosinophilic/basophilic-related differentiation marker the Charcot-Leyden crystal. Treatment of HMC-1 with conditioned media from the human T-cell line Mo, retinoic acid or DMSO induced only moderate changes in the surface or intracellular expression of the studied markers. The agents tested neither induced any of the monocyte/granulocyte markers examined, nor expression of the Fc epsilon RI alpha-chain.
Nilsson
G
G
Department of Pathology, University of Uppsala, Sweden.
Blom
T
T
Kusche-Gullberg
M
M
Kjellén
L
L
Butterfield
J H
JH
Sundström
C
C
Nilsson
K
K
Hellman
L
L
D000595:Amino Acid Sequence; D000943:Antigens, Differentiation / Q000096:biosynthesis; D015152:Blotting, Northern; D002454:Cell Differentiation / Q000187:drug effects; D053818:Chymases; D005434:Flow Cytometry; D006025:Glycosaminoglycans / Q000032:analysis; D006632:Histamine / Q000032:analysis; D006801:Humans; D007124:Immunoenzyme Techniques; D016130:Immunophenotyping; D008407:Mast Cells / Q000737:chemistry* / Q000276:immunology*; D008969:Molecular Sequence Data; D012697:Serine Endopeptidases / Q000032:analysis; D053802:Tryptases; D014407:Tumor Cells, Cultured / Q000737:chemistry* / Q000276:immunology*; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis
D000943:Antigens, Differentiation; D006025:Glycosaminoglycans; D014409:Tumor Necrosis Factor-alpha; D006632:Histamine; D012697:Serine Endopeptidases; C083294:chymase 2; D053818:Chymases; D053802:Tryptases