Association of IgG Fc receptor II with tyrosine kinases in the human basophilic leukemia cell line KU812F

Makoto Fujii, Yasushi Tanimoto, Minoru Takata, Kazushi Takao, Noboru Hamada, Toshimitsu Suwaki, Noriko Kawata, Kiyoshi Takahashi, Mine Harada, Mitsune Tanimoto

Research output: Contribution to journalArticle

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Abstract

Background: We previously reported that cross-linking of IgG Fc receptor II (FγRII) induces intracellular calcium mobilization, but not histamine release in human basophils. To clarify functional activities of FcγRII on human basophils, we analyzed the FcγRII-mediated signaling events in the human basophilic leukemia cell line KU812F. Methods: Flow cytometric methods were used to investigate the effect on intracellular calcium mobilization of cross-linking of FγRII. KU812F cells were pre-incubated with anti-FcγRII monoclonal antibody (IV.3). After the addition of various concentrations of the tyrosine kinase inhibitor genistein or buffer alone, cells were stimulated with goat antimouse IgG F(ab′)2 (GAM) and analyzed with the flow cytometer. Next, in order to test the signaling events after cross-linking of FcγRII, we examined tyrosine kinase activity. The time-course of tyrosine phosphorylation after cross-linking of FcγRII and the effect of genistein on this tyrosine phosphorylation were tested by immunoblotting. Immunoprecipitation was also performed to identify the type of tyrosine kinase associated with signal transduction of FcγRII. Results: The tyrosine kinase inhibitor genistein inhibited intracellular calcium mobilization caused by cross-linking of FcγRII in a dose-dependent manner. Rapid tyrosine phosphorylation after FcγRII cross-linking was shown by immunoblot analysis and this phosphorylation was inhibited by genistein. Furthermore, tyrosine phosphorylation of Lyn and Syk was observed upon cross-linking of FcγRII. Conclusions: Tyrosine phosphorylation is necessary for the signaling pathway through FcγRII and tyrosine phosphorylation of Lyn and Syk, at least, is actively involved in this signal transduction.

Original languageEnglish
Pages (from-to)149-154
Number of pages6
JournalAllergology International
Volume52
Issue number3
DOIs
Publication statusPublished - Sep 2003

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IgG Receptors
Fc Receptors
Protein-Tyrosine Kinases
Leukemia
Tyrosine
Phosphorylation
Genistein
Cell Line
Basophils
Calcium
Signal Transduction
Histamine Release
Immunoprecipitation
Immunoblotting
Goats
Buffers
Immunoglobulin G
Monoclonal Antibodies

Keywords

  • Basophilic leukemia cell line
  • IgG Fc receptor II
  • Intracellular calcium mobilization
  • Signal transduction
  • Tyrosine kinase

ASJC Scopus subject areas

  • Immunology and Allergy

Cite this

Association of IgG Fc receptor II with tyrosine kinases in the human basophilic leukemia cell line KU812F. / Fujii, Makoto; Tanimoto, Yasushi; Takata, Minoru; Takao, Kazushi; Hamada, Noboru; Suwaki, Toshimitsu; Kawata, Noriko; Takahashi, Kiyoshi; Harada, Mine; Tanimoto, Mitsune.

In: Allergology International, Vol. 52, No. 3, 09.2003, p. 149-154.

Research output: Contribution to journalArticle

Fujii, M, Tanimoto, Y, Takata, M, Takao, K, Hamada, N, Suwaki, T, Kawata, N, Takahashi, K, Harada, M & Tanimoto, M 2003, 'Association of IgG Fc receptor II with tyrosine kinases in the human basophilic leukemia cell line KU812F', Allergology International, vol. 52, no. 3, pp. 149-154. https://doi.org/10.1046/j.1440-1592.2003.00291.x
Fujii, Makoto ; Tanimoto, Yasushi ; Takata, Minoru ; Takao, Kazushi ; Hamada, Noboru ; Suwaki, Toshimitsu ; Kawata, Noriko ; Takahashi, Kiyoshi ; Harada, Mine ; Tanimoto, Mitsune. / Association of IgG Fc receptor II with tyrosine kinases in the human basophilic leukemia cell line KU812F. In: Allergology International. 2003 ; Vol. 52, No. 3. pp. 149-154.
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abstract = "Background: We previously reported that cross-linking of IgG Fc receptor II (FγRII) induces intracellular calcium mobilization, but not histamine release in human basophils. To clarify functional activities of FcγRII on human basophils, we analyzed the FcγRII-mediated signaling events in the human basophilic leukemia cell line KU812F. Methods: Flow cytometric methods were used to investigate the effect on intracellular calcium mobilization of cross-linking of FγRII. KU812F cells were pre-incubated with anti-FcγRII monoclonal antibody (IV.3). After the addition of various concentrations of the tyrosine kinase inhibitor genistein or buffer alone, cells were stimulated with goat antimouse IgG F(ab′)2 (GAM) and analyzed with the flow cytometer. Next, in order to test the signaling events after cross-linking of FcγRII, we examined tyrosine kinase activity. The time-course of tyrosine phosphorylation after cross-linking of FcγRII and the effect of genistein on this tyrosine phosphorylation were tested by immunoblotting. Immunoprecipitation was also performed to identify the type of tyrosine kinase associated with signal transduction of FcγRII. Results: The tyrosine kinase inhibitor genistein inhibited intracellular calcium mobilization caused by cross-linking of FcγRII in a dose-dependent manner. Rapid tyrosine phosphorylation after FcγRII cross-linking was shown by immunoblot analysis and this phosphorylation was inhibited by genistein. Furthermore, tyrosine phosphorylation of Lyn and Syk was observed upon cross-linking of FcγRII. Conclusions: Tyrosine phosphorylation is necessary for the signaling pathway through FcγRII and tyrosine phosphorylation of Lyn and Syk, at least, is actively involved in this signal transduction.",
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T1 - Association of IgG Fc receptor II with tyrosine kinases in the human basophilic leukemia cell line KU812F

AU - Fujii, Makoto

AU - Tanimoto, Yasushi

AU - Takata, Minoru

AU - Takao, Kazushi

AU - Hamada, Noboru

AU - Suwaki, Toshimitsu

AU - Kawata, Noriko

AU - Takahashi, Kiyoshi

AU - Harada, Mine

AU - Tanimoto, Mitsune

PY - 2003/9

Y1 - 2003/9

N2 - Background: We previously reported that cross-linking of IgG Fc receptor II (FγRII) induces intracellular calcium mobilization, but not histamine release in human basophils. To clarify functional activities of FcγRII on human basophils, we analyzed the FcγRII-mediated signaling events in the human basophilic leukemia cell line KU812F. Methods: Flow cytometric methods were used to investigate the effect on intracellular calcium mobilization of cross-linking of FγRII. KU812F cells were pre-incubated with anti-FcγRII monoclonal antibody (IV.3). After the addition of various concentrations of the tyrosine kinase inhibitor genistein or buffer alone, cells were stimulated with goat antimouse IgG F(ab′)2 (GAM) and analyzed with the flow cytometer. Next, in order to test the signaling events after cross-linking of FcγRII, we examined tyrosine kinase activity. The time-course of tyrosine phosphorylation after cross-linking of FcγRII and the effect of genistein on this tyrosine phosphorylation were tested by immunoblotting. Immunoprecipitation was also performed to identify the type of tyrosine kinase associated with signal transduction of FcγRII. Results: The tyrosine kinase inhibitor genistein inhibited intracellular calcium mobilization caused by cross-linking of FcγRII in a dose-dependent manner. Rapid tyrosine phosphorylation after FcγRII cross-linking was shown by immunoblot analysis and this phosphorylation was inhibited by genistein. Furthermore, tyrosine phosphorylation of Lyn and Syk was observed upon cross-linking of FcγRII. Conclusions: Tyrosine phosphorylation is necessary for the signaling pathway through FcγRII and tyrosine phosphorylation of Lyn and Syk, at least, is actively involved in this signal transduction.

AB - Background: We previously reported that cross-linking of IgG Fc receptor II (FγRII) induces intracellular calcium mobilization, but not histamine release in human basophils. To clarify functional activities of FcγRII on human basophils, we analyzed the FcγRII-mediated signaling events in the human basophilic leukemia cell line KU812F. Methods: Flow cytometric methods were used to investigate the effect on intracellular calcium mobilization of cross-linking of FγRII. KU812F cells were pre-incubated with anti-FcγRII monoclonal antibody (IV.3). After the addition of various concentrations of the tyrosine kinase inhibitor genistein or buffer alone, cells were stimulated with goat antimouse IgG F(ab′)2 (GAM) and analyzed with the flow cytometer. Next, in order to test the signaling events after cross-linking of FcγRII, we examined tyrosine kinase activity. The time-course of tyrosine phosphorylation after cross-linking of FcγRII and the effect of genistein on this tyrosine phosphorylation were tested by immunoblotting. Immunoprecipitation was also performed to identify the type of tyrosine kinase associated with signal transduction of FcγRII. Results: The tyrosine kinase inhibitor genistein inhibited intracellular calcium mobilization caused by cross-linking of FcγRII in a dose-dependent manner. Rapid tyrosine phosphorylation after FcγRII cross-linking was shown by immunoblot analysis and this phosphorylation was inhibited by genistein. Furthermore, tyrosine phosphorylation of Lyn and Syk was observed upon cross-linking of FcγRII. Conclusions: Tyrosine phosphorylation is necessary for the signaling pathway through FcγRII and tyrosine phosphorylation of Lyn and Syk, at least, is actively involved in this signal transduction.

KW - Basophilic leukemia cell line

KW - IgG Fc receptor II

KW - Intracellular calcium mobilization

KW - Signal transduction

KW - Tyrosine kinase

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