Presentation is loading. Please wait.

Presentation is loading. Please wait.

Factors affecting the cytokine production of human T cells stimulated by different modes of activation  Yoshio Harada, MDa, Sumiko Watanabe, PhDa, Hans.

Published byFanny Atmadjaja Modified over 5 years ago

Similar presentations

Presentation on theme: "Factors affecting the cytokine production of human T cells stimulated by different modes of activation  Yoshio Harada, MDa, Sumiko Watanabe, PhDa, Hans."— Presentation transcript:

1 Factors affecting the cytokine production of human T cells stimulated by different modes of activation  Yoshio Harada, MDa, Sumiko Watanabe, PhDa, Hans Yssel, PhDb, Ken-ichi Arai, MD, PhDa  Journal of Allergy and Clinical Immunology  Volume 98, Issue 6, Pages S161-S173 (December 1996) DOI: /S (96) Copyright © 1996 Mosby, Inc. Terms and Conditions

2 FIG. 1 Profile of cytokine production of human T-cell clones SP-B21 (TH0-like) and TA20 and TA23 (TH1-like). Cells (2 × 105) were stimulated with (A) αCD3-mAb (10 μg/ml, coated on 96-well plate) or (B) PMA (1 ng/ml) and A23187 (500 ng/ml). After 48 hours of incubation at 37° C, supernatants were collected, and levels of cytokines were determined by cytokine-specific ELISA. Journal of Allergy and Clinical Immunology  , S161-S173DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

3 FIG. 1 Profile of cytokine production of human T-cell clones SP-B21 (TH0-like) and TA20 and TA23 (TH1-like). Cells (2 × 105) were stimulated with (A) αCD3-mAb (10 μg/ml, coated on 96-well plate) or (B) PMA (1 ng/ml) and A23187 (500 ng/ml). After 48 hours of incubation at 37° C, supernatants were collected, and levels of cytokines were determined by cytokine-specific ELISA. Journal of Allergy and Clinical Immunology  , S161-S173DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

4 FIG. 2 Effects of herbimycin on cytokine production of human T-cell clone SP-B21. Cells (2 × 105) were stimulated with soluble αCD3-mAb (1 μg/ml) or PMA (1 ng/ml)/A23187 (500 ng/ml) in presence of various concentrations of herbimycin. As described in legend to Fig. 1, levels of IFN-γ (A), IL-4 (B), and IL-5 (C) were measured by cytokine-specific ELISA. Results are expressed as percentage of control to facilitate comparison. Control levels of cytokine production in response to αCD3-mAb and PMA/A23187, respectively, were as follows: IFN-γ, 1.7 and 45.9 ng/ml; IL-4, 0.31 and 4.0 ng/ml; IL-5, 3.9 and 8.7 ng/ml. Journal of Allergy and Clinical Immunology  , S161-S173DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

5 FIG. 2 Effects of herbimycin on cytokine production of human T-cell clone SP-B21. Cells (2 × 105) were stimulated with soluble αCD3-mAb (1 μg/ml) or PMA (1 ng/ml)/A23187 (500 ng/ml) in presence of various concentrations of herbimycin. As described in legend to Fig. 1, levels of IFN-γ (A), IL-4 (B), and IL-5 (C) were measured by cytokine-specific ELISA. Results are expressed as percentage of control to facilitate comparison. Control levels of cytokine production in response to αCD3-mAb and PMA/A23187, respectively, were as follows: IFN-γ, 1.7 and 45.9 ng/ml; IL-4, 0.31 and 4.0 ng/ml; IL-5, 3.9 and 8.7 ng/ml. Journal of Allergy and Clinical Immunology  , S161-S173DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

6 FIG. 2 Effects of herbimycin on cytokine production of human T-cell clone SP-B21. Cells (2 × 105) were stimulated with soluble αCD3-mAb (1 μg/ml) or PMA (1 ng/ml)/A23187 (500 ng/ml) in presence of various concentrations of herbimycin. As described in legend to Fig. 1, levels of IFN-γ (A), IL-4 (B), and IL-5 (C) were measured by cytokine-specific ELISA. Results are expressed as percentage of control to facilitate comparison. Control levels of cytokine production in response to αCD3-mAb and PMA/A23187, respectively, were as follows: IFN-γ, 1.7 and 45.9 ng/ml; IL-4, 0.31 and 4.0 ng/ml; IL-5, 3.9 and 8.7 ng/ml. Journal of Allergy and Clinical Immunology  , S161-S173DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

7 FIG. 3 Effects of CsA on cytokine production of SP-B21. Cells (2 × 105) were stimulated with αCD3-mAb (1 μg/ml) (A) or PMA (1 ng/ml)/αCD3-mAb (1 μg/ml) (B) in presence of various concentrations of CsA. As described in legend to Fig. 1, levels of IFN-γ, IL-4, and IL-5 were measured by cytokine-specific ELISA. Results are expressed as percentage of control to facilitate comparison. Control levels of cytokine production in response to αCD3-mAb and PMA/A23187, respectively, were as follows: IFN-γ, 1.7 and 19.7 ng/ml; IL-4, 0.31 and 5.2 ng/ml; IL-5, 3.9 and 9.3 ng/ml. Journal of Allergy and Clinical Immunology  , S161-S173DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

8 FIG. 3 Effects of CsA on cytokine production of SP-B21. Cells (2 × 105) were stimulated with αCD3-mAb (1 μg/ml) (A) or PMA (1 ng/ml)/αCD3-mAb (1 μg/ml) (B) in presence of various concentrations of CsA. As described in legend to Fig. 1, levels of IFN-γ, IL-4, and IL-5 were measured by cytokine-specific ELISA. Results are expressed as percentage of control to facilitate comparison. Control levels of cytokine production in response to αCD3-mAb and PMA/A23187, respectively, were as follows: IFN-γ, 1.7 and 19.7 ng/ml; IL-4, 0.31 and 5.2 ng/ml; IL-5, 3.9 and 9.3 ng/ml. Journal of Allergy and Clinical Immunology  , S161-S173DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

9 FIG. 4 Effects of A23187 on cytokine production of SP-B21. Cells (2 × 105) were stimulated with αCD3-mAb (1 μg/ml), PMA (1 ng/ml), or αCD3-mAb (1 μg/ml)/PMA (1 ng/ml) in presence of various concentrations of A As described in legend to Fig. 1, levels of IL-10 (A), IFN-γ (B), and IL-4 (C) were measured by cytokine-specific ELISA. Journal of Allergy and Clinical Immunology  , S161-S173DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

10 FIG. 4 Effects of A23187 on cytokine production of SP-B21. Cells (2 × 105) were stimulated with αCD3-mAb (1 μg/ml), PMA (1 ng/ml), or αCD3-mAb (1 μg/ml)/PMA (1 ng/ml) in presence of various concentrations of A As described in legend to Fig. 1, levels of IL-10 (A), IFN-γ (B), and IL-4 (C) were measured by cytokine-specific ELISA. Journal of Allergy and Clinical Immunology  , S161-S173DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

11 FIG. 4 Effects of A23187 on cytokine production of SP-B21. Cells (2 × 105) were stimulated with αCD3-mAb (1 μg/ml), PMA (1 ng/ml), or αCD3-mAb (1 μg/ml)/PMA (1 ng/ml) in presence of various concentrations of A As described in legend to Fig. 1, levels of IL-10 (A), IFN-γ (B), and IL-4 (C) were measured by cytokine-specific ELISA. Journal of Allergy and Clinical Immunology  , S161-S173DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

12 FIG. 5 Effects of αIL-2Rα mAb (A) or TGF-β (B) on proliferation of SP-B21. Cells (2 × 104) were stimulated with soluble αCD3-mAb (1 μg/ml), PMA (1 ng/ml), or PMA (1 ng/ml)/A23187 (500 ng/ml) in presence or absence of αIL-2Rα mAb or TGF-β. [3H]Thymidine incorporation was measured from triplicate samples. Journal of Allergy and Clinical Immunology  , S161-S173DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

13 FIG. 5 Effects of αIL-2Rα mAb (A) or TGF-β (B) on proliferation of SP-B21. Cells (2 × 104) were stimulated with soluble αCD3-mAb (1 μg/ml), PMA (1 ng/ml), or PMA (1 ng/ml)/A23187 (500 ng/ml) in presence or absence of αIL-2Rα mAb or TGF-β. [3H]Thymidine incorporation was measured from triplicate samples. Journal of Allergy and Clinical Immunology  , S161-S173DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

14 FIG. 6 Effects of PGE2 on cytokine production and proliferation of SP-B21. Cells (2 × 105) were stimulated with αCD3-mAb (1 μg/ml) or PMA (1 ng/ml)/A23187 (500 ng/ml) and with or without PGE2 (10 μmol/L). Total RNA was isolated after 4 hours of incubation with or without PGE2. Cytokine mRNA was analyzed with reverse-transcription PCR. Journal of Allergy and Clinical Immunology  , S161-S173DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

15 FIG. 7 Effects of dBcAMP on cytokine production of TH0-like (SP-B21) and TH1-like (TA20 and TA23) human T-cell clones. Cells (2 × 105) were stimulated with αCD3-mAb (1 μg/ml) (A and C) or PMA (1 ng/ml)/A23187 (500 ng/ml) (B and D) and with or without dBcAMP (1 mmol/L). As described in legend to Fig. 1, levels of IFN-γ (A and B) and IL-4 (C and D) were measured by cytokine-specific ELISA. Journal of Allergy and Clinical Immunology  , S161-S173DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

16 FIG. 7 Effects of dBcAMP on cytokine production of TH0-like (SP-B21) and TH1-like (TA20 and TA23) human T-cell clones. Cells (2 × 105) were stimulated with αCD3-mAb (1 μg/ml) (A and C) or PMA (1 ng/ml)/A23187 (500 ng/ml) (B and D) and with or without dBcAMP (1 mmol/L). As described in legend to Fig. 1, levels of IFN-γ (A and B) and IL-4 (C and D) were measured by cytokine-specific ELISA. Journal of Allergy and Clinical Immunology  , S161-S173DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

17 FIG. 7 Effects of dBcAMP on cytokine production of TH0-like (SP-B21) and TH1-like (TA20 and TA23) human T-cell clones. Cells (2 × 105) were stimulated with αCD3-mAb (1 μg/ml) (A and C) or PMA (1 ng/ml)/A23187 (500 ng/ml) (B and D) and with or without dBcAMP (1 mmol/L). As described in legend to Fig. 1, levels of IFN-γ (A and B) and IL-4 (C and D) were measured by cytokine-specific ELISA. Journal of Allergy and Clinical Immunology  , S161-S173DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

18 FIG. 7 Effects of dBcAMP on cytokine production of TH0-like (SP-B21) and TH1-like (TA20 and TA23) human T-cell clones. Cells (2 × 105) were stimulated with αCD3-mAb (1 μg/ml) (A and C) or PMA (1 ng/ml)/A23187 (500 ng/ml) (B and D) and with or without dBcAMP (1 mmol/L). As described in legend to Fig. 1, levels of IFN-γ (A and B) and IL-4 (C and D) were measured by cytokine-specific ELISA. Journal of Allergy and Clinical Immunology  , S161-S173DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

19 FIG. 8 Electrophoretic mobility shift assay of NF-κB complex. SP-B21 cells (5 × 106) were stimulated with αCD3-mAb (1 μg/ml) or PMA (1 ng/ml)/A23187 (500 ng/ml) and with or without (A) PGE2 (10 μmol/L) or (B) dBcAMP (1 mmol/L). At 4 hours after stimulation, nuclear extracts prepared from these cells were incubated with IgκB oligonucleotide labeled with [α-32P]dGTP at room temperature for 15 minutes, and DNA–protein complexes were electrophoresed at 100 V on 4% polyacrylamide gel. Journal of Allergy and Clinical Immunology  , S161-S173DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

20 FIG. 8 Electrophoretic mobility shift assay of NF-κB complex. SP-B21 cells (5 × 106) were stimulated with αCD3-mAb (1 μg/ml) or PMA (1 ng/ml)/A23187 (500 ng/ml) and with or without (A) PGE2 (10 μmol/L) or (B) dBcAMP (1 mmol/L). At 4 hours after stimulation, nuclear extracts prepared from these cells were incubated with IgκB oligonucleotide labeled with [α-32P]dGTP at room temperature for 15 minutes, and DNA–protein complexes were electrophoresed at 100 V on 4% polyacrylamide gel. Journal of Allergy and Clinical Immunology  , S161-S173DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

21 FIG. 9 Electrophoretic mobility shift assay of complex bound to NF-AT motif. SP-B21 cells (5 × 106) were stimulated with αCD3-mAb (1 μg/ml) and with or without PGE2 (10 μmol/L). Cells were harvested 4 hours after stimulation, and nuclear extracts were prepared and electrophoretic mobility shift assays performed as described in legend to Fig. 8. Journal of Allergy and Clinical Immunology  , S161-S173DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

Download ppt "Factors affecting the cytokine production of human T cells stimulated by different modes of activation  Yoshio Harada, MDa, Sumiko Watanabe, PhDa, Hans."

Similar presentations

Proinflammatory cytokine–induced and chemical mediator–induced IL-8 expression in human bronchial epithelial cells through p38 mitogen-activated protein.

Proinflammatory cytokine–induced and chemical mediator–induced IL-8 expression in human bronchial epithelial cells through p38 mitogen-activated protein.
Immunopathogenesis of human gastrointestinal infection by Anisakis simplex  Victoria del Pozo, PhDa, Ignacio Arrieta, MDa, Teresa Tuñon, MD, PhDb, Isabel.

Immunopathogenesis of human gastrointestinal infection by Anisakis simplex  Victoria del Pozo, PhDa, Ignacio Arrieta, MDa, Teresa Tuñon, MD, PhDb, Isabel.
Glucocorticoids inhibit chemokine generation by human eosinophils

Glucocorticoids inhibit chemokine generation by human eosinophils
Interleukin-4 receptor expression by human B cells: Functional analysis with a human interleukin-4 toxin, DAB389IL-4  Haifa H. Jabara, MSca, Donata Vercelli,

Interleukin-4 receptor expression by human B cells: Functional analysis with a human interleukin-4 toxin, DAB389IL-4  Haifa H. Jabara, MSca, Donata Vercelli,
Activation of mast cells by double-stranded RNA: evidence for activation through Toll- like receptor 3  Marianna Kulka, PhD, Lena Alexopoulou, PhD, Richard.

Activation of mast cells by double-stranded RNA: evidence for activation through Toll- like receptor 3  Marianna Kulka, PhD, Lena Alexopoulou, PhD, Richard.
Interleukin-4 receptor expression by human B cells: Functional analysis with a human interleukin-4 toxin, DAB389IL-4  Haifa H. Jabara, MSca, Donata Vercelli,

Interleukin-4 receptor expression by human B cells: Functional analysis with a human interleukin-4 toxin, DAB389IL-4  Haifa H. Jabara, MSca, Donata Vercelli,
Role of γ/δ T cells in a patient with CD4+ CD3– lymphocytosis, hypereosinophilia, and high levels of IgE  Ilan Bank, MDa,d, Avner Reshef, MDb, Miriam.

Role of γ/δ T cells in a patient with CD4+ CD3– lymphocytosis, hypereosinophilia, and high levels of IgE  Ilan Bank, MDa,d, Avner Reshef, MDb, Miriam.
Heat-killed Lactobacillus plantarum L-137 suppresses naturally fed antigen–specific IgE production by stimulation of IL-12 production in mice  Shinji.

Heat-killed Lactobacillus plantarum L-137 suppresses naturally fed antigen–specific IgE production by stimulation of IL-12 production in mice  Shinji.
The environmental pollutant pyrene induces the production of IL-4

The environmental pollutant pyrene induces the production of IL-4
Nerve growth factor is preformed in and activates human peripheral blood eosinophils  Abraham Solomon, MDa,b, Luigi Aloe, PhDc, Jacob Pe’er, MDa, Joseph.

Nerve growth factor is preformed in and activates human peripheral blood eosinophils  Abraham Solomon, MDa,b, Luigi Aloe, PhDc, Jacob Pe’er, MDa, Joseph.
Modulation by aspirin of nuclear phospho–signal transducer and activator of transcription 6 expression: Possible role in therapeutic benefit associated.

Modulation by aspirin of nuclear phospho–signal transducer and activator of transcription 6 expression: Possible role in therapeutic benefit associated.
Roles of the cytoplasmic domains of the α and β subunits of human granulocyte- macrophage colony-stimulating factor receptor  Akihiko Muto, PhDa, Sumiko.

Roles of the cytoplasmic domains of the α and β subunits of human granulocyte- macrophage colony-stimulating factor receptor  Akihiko Muto, PhDa, Sumiko.
Epigenetic regulation of established human type 1 versus type 2 cytokine responses  Ruey-Chyi Su, PhD, Allan B. Becker, MD, Anita L. Kozyrskyj, PhD, Kent.

Epigenetic regulation of established human type 1 versus type 2 cytokine responses  Ruey-Chyi Su, PhD, Allan B. Becker, MD, Anita L. Kozyrskyj, PhD, Kent.
Christine M. Braun, BA, Shau-Ku Huang, PhD, Gregory G

Christine M. Braun, BA, Shau-Ku Huang, PhD, Gregory G
Eosinophil apoptosis caused by theophylline, glucocorticoids, and macrolides after stimulation with IL-5  Tetsuya Adachi, MDa, Shinji Motojima, MDb, Akira.

Eosinophil apoptosis caused by theophylline, glucocorticoids, and macrolides after stimulation with IL-5  Tetsuya Adachi, MDa, Shinji Motojima, MDb, Akira.
Signal transduction pathways triggered by the FcϵRIIb receptor (CD23) in human monocytes lead to nuclear factor-κB activation  Rosa M. Ten, MD, PhDa,

Signal transduction pathways triggered by the FcϵRIIb receptor (CD23) in human monocytes lead to nuclear factor-κB activation  Rosa M. Ten, MD, PhDa,
The inhibitory effects of topically active glucocorticoids on IL-4, IL-5, and interferon-γ production by cultured primary CD4+ T cells  Shelby P. Umland,

The inhibitory effects of topically active glucocorticoids on IL-4, IL-5, and interferon-γ production by cultured primary CD4+ T cells  Shelby P. Umland,
CD40-mediated p38 mitogen-activated protein kinase activation is required for immunoglobulin class switch recombination to IgE  Ke Zhang, MD, PhD, Ling.

CD40-mediated p38 mitogen-activated protein kinase activation is required for immunoglobulin class switch recombination to IgE  Ke Zhang, MD, PhD, Ling.
Estrogen enhances immunoglobulin production by human PBMCs

Estrogen enhances immunoglobulin production by human PBMCs
Prostaglandin E2 control of T cell cytokine production is functionally related to the reduced lymphocyte proliferation in atopic dermatitis  Sai Chan,

Prostaglandin E2 control of T cell cytokine production is functionally related to the reduced lymphocyte proliferation in atopic dermatitis  Sai Chan,

Similar presentations

Ads by Google