Cultured human mast cells derived from umbilical cord blood cells in the presence of stem cell factor and interleukin-6 cannot be a model of human skin mast cells: fluorescence microscopic analysis of intracellular calcium ion mobilization

In the present study, we attempted to clarify the effects of interleukin-6 (IL-6) on the growth and properties of human mast cells using cultured mast cells selectively generated by stem cell factor (SCF) from CD34+ cord blood cells. The addition of IL-6 to cultures containing mast cells resulted in a substantial reduction of the number of progenies grown by SCF in the liquid culture. This IL-6–mediated inhibition of mast cell growth may be due in part to the suppression at the precursor level, according to the results of a clonal cell culture assay. Moreover, a flow cytometric analysis showed that the cultured mast cells grown in the presence of SCF+IL-6 had decreased c-kit expression. The exposure of cultured mast cells to SCF+IL-6 also caused substantial increases in the cell size, frequency of chymase-positive cells, and intracellular histamine level compared with the values obtained with SCF alone. The flow cytometric analysis showed low but significant levels of expression of IL-6 receptor (IL-6R) and gp130 on the cultured mast cells grown with SCF. The addition of either anti–IL-6R antibody or anti-gp130 antibody abrogated the biological functions of IL-6. Although IL-4 exerted an effect similar to that of IL-6 on the cultured mast cells under stimulation with SCF, the results of comparative experiments suggest that the two cytokines use different regulatory mechanisms. Taken together, the present findings suggest that IL-6 modulates SCF-dependent human mast cell development directly via an IL-6R-gp130 system.

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An Initial Signal of Activation of Rat Peritoneal Mast Cells Stimulated by Datura stramonium Agglutinin: A Confocal Fluorescence Microscopic Analysis of Intracellular Calcium Ion and Cytoskeletal Assembly

The Japanese Journal of Pharmacology ◽

10.1254/jjp.66.205 ◽

1994 ◽

Vol 66 (2) ◽

pp. 205-212 ◽

Cited By ~ 5

Author(s):

Tamiko Suzuki-Nishimura ◽

Tadahide Furuno ◽

Masaatsu K. Uchida ◽

Mamoru Nakanishi

Keyword(s):

Mast Cells ◽

Calcium Ion ◽

Microscopic Analysis ◽

Datura Stramonium ◽

Initial Signal ◽

Peritoneal Mast Cells ◽

Confocal Fluorescence ◽

Intracellular Calcium Ion ◽

Rat Peritoneal Mast Cells ◽

Fluorescence Microscopic Analysis

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Interleukin-6 Directly Modulates Stem Cell Factor-Dependent Development of Human Mast Cells Derived From CD34+Cord Blood Cells

Blood ◽

10.1182/blood.v94.2.496 ◽

1999 ◽

Vol 94 (2) ◽

pp. 496-508 ◽

Cited By ~ 64

Author(s):

Tatsuya Kinoshita ◽

Nobukuni Sawai ◽

Eiko Hidaka ◽

Tetsuji Yamashita ◽

Kenichi Koike

Keyword(s):

Mast Cell ◽

Stem Cell ◽

Mast Cells ◽

Cord Blood ◽

Interleukin 6 ◽

Blood Cells ◽

Stem Cell Factor ◽

Flow Cytometric Analysis ◽

Flow Cytometric ◽

Cord Blood Cells

Abstract In the present study, we attempted to clarify the effects of interleukin-6 (IL-6) on the growth and properties of human mast cells using cultured mast cells selectively generated by stem cell factor (SCF) from CD34+ cord blood cells. The addition of IL-6 to cultures containing mast cells resulted in a substantial reduction of the number of progenies grown by SCF in the liquid culture. This IL-6–mediated inhibition of mast cell growth may be due in part to the suppression at the precursor level, according to the results of a clonal cell culture assay. Moreover, a flow cytometric analysis showed that the cultured mast cells grown in the presence of SCF+IL-6 had decreased c-kit expression. The exposure of cultured mast cells to SCF+IL-6 also caused substantial increases in the cell size, frequency of chymase-positive cells, and intracellular histamine level compared with the values obtained with SCF alone. The flow cytometric analysis showed low but significant levels of expression of IL-6 receptor (IL-6R) and gp130 on the cultured mast cells grown with SCF. The addition of either anti–IL-6R antibody or anti-gp130 antibody abrogated the biological functions of IL-6. Although IL-4 exerted an effect similar to that of IL-6 on the cultured mast cells under stimulation with SCF, the results of comparative experiments suggest that the two cytokines use different regulatory mechanisms. Taken together, the present findings suggest that IL-6 modulates SCF-dependent human mast cell development directly via an IL-6R-gp130 system.

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