Chemokines induce eosinophil degranulation through CCR-3

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Presentation transcript:

Chemokines induce eosinophil degranulation through CCR-3 Takao Fujisawa, MDa, Yoshiko Kato, MSa,b, Hiroyuki Nagase, MDf, Jun Atsuta, MDa, Akihiko Terada, MDa, Kosei Iguchi, MDa, Hitoshi Kamiya, MDa, Yutaka Morita, MDf, Motoji Kitaura, MSc, Hiroshi Kawasaki, MD, PhDd, Osamu Yoshie, MDe, Koichi Hirai, MD, PhDg Journal of Allergy and Clinical Immunology Volume 106, Issue 3, Pages 507-513 (September 2000) DOI: 10.1067/mai.2000.108311 Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 1 Calcium influx to eosinophils induced by chemokines. Eosinophils were stimulated with a panel of chemokines at a concentration of 333 ng/mL, and calcium influx was measured as described in the “Methods” section. The data shown are representative of 2 or 4 independent analyses from different donors, each showing similar results. Journal of Allergy and Clinical Immunology 2000 106, 507-513DOI: (10.1067/mai.2000.108311) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 2 Chemokine-induced eosinophil degranulation. Eosinophils at 1 × 106/mL were pretreated with 5 μg/mL CB for 5 minutes and incubated with various chemokines at 1 × 10–7 mol/L in the absence (A) or presence (B) of IL-5 at 2.5 ng/mL for 5 hours. Results are expressed as means ± SEM of the percentage of EDN released in 4 duplicate experiments. *P < .05 (significantly different from control release without chemokines). SCM, Single C motif; TARC, thymus and activation-regulated chemokine; LARC, liver and activation-regulated chemokine; SLC, secondary lymphoid-tissue chemokine; PARC, pulmonary and activation-regulated chemokine. Journal of Allergy and Clinical Immunology 2000 106, 507-513DOI: (10.1067/mai.2000.108311) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 3 Kinetics of eosinophil degranulation induced by eotaxin and IL-5. Eosinophils at 1 × 106/mL were treated with 5 μg/mL CB for 5 minutes and then incubated with eotaxin at 1 × 10–7 mol/L and IL-5 at 2.5 ng/mL, with IL-5 at 2.5 ng/mL, or with medium, respectively, for the indicated hours. Levels of EDN in the supernatants and cell lysates were measured with RIA. Results are expressed as means ± SEM of the percentage of EDN released in 3 duplicate experiments. Journal of Allergy and Clinical Immunology 2000 106, 507-513DOI: (10.1067/mai.2000.108311) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 4 Eosinophil degranulation by eotaxin and RANTES. Eosinophils at 1 × 106/mL were treated with 5 μg/mL CB for 5 minutes and then incubated with varying concentrations of eotaxin (A) or RANTES (B) with or without IL-5 at 2.5 ng/mL for 5 hours. Results are expressed as means ± SEM of the percentage of EDN released in 4 duplicate experiments. *P < .005 (significantly different from control release without chemokines). Journal of Allergy and Clinical Immunology 2000 106, 507-513DOI: (10.1067/mai.2000.108311) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 5 Inhibition of chemokine-induced eosinophil degranulation by an anti-CCR3 antibody. Eosinophils at 1 × 106/mL were pretreated with 5 μg/mL CB for 5 minutes and stimulated with eotaxin at 1 × 10–7 mol/L and IL-5 at 2.5 ng/mL in the presence of anti-CCR1, anti-CCR3, or isotype-matched IgG at 10 μg/mL for 5 hours (A) . Similar experiments were performed with RANTES at 1 × 10–7 mol/L and IL-5 at 2.5 ng/mL (B) or with C5a at 5 × 10–8 mol/L (C) as stimulators. Results are expressed as means ± SEM of the percentage of EDN released in 4 duplicate experiments. *P < .05 (significantly different from IgG1 control by the paired t test). Journal of Allergy and Clinical Immunology 2000 106, 507-513DOI: (10.1067/mai.2000.108311) Copyright © 2000 Mosby, Inc. Terms and Conditions