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Mast cell–derived prostaglandin D2 attenuates anaphylactic reactions in mice  Tatsuro Nakamura, DVM, PhD, Yuki Fujiwara, BSc, Ryota Yamada, BSc, Wataru.

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Presentation on theme: "Mast cell–derived prostaglandin D2 attenuates anaphylactic reactions in mice  Tatsuro Nakamura, DVM, PhD, Yuki Fujiwara, BSc, Ryota Yamada, BSc, Wataru."— Presentation transcript:

1 Mast cell–derived prostaglandin D2 attenuates anaphylactic reactions in mice 
Tatsuro Nakamura, DVM, PhD, Yuki Fujiwara, BSc, Ryota Yamada, BSc, Wataru Fujii, PhD, Taiki Hamabata, BSc, Monica Yunkyung Lee, PhD, Shingo Maeda, DVM, PhD, Kosuke Aritake, PhD, Axel Roers, MD, PhD, William C. Sessa, PhD, Masataka Nakamura, PhD, Yoshihiro Urade, PhD, Takahisa Murata, DVM, PhD  Journal of Allergy and Clinical Immunology  Volume 140, Issue 2, Pages e9 (August 2017) DOI: /j.jaci Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Systemic and mast cell–specific H-PGDS deficiency exacerbates anaphylactic reactions. A-G, Antigen/antibody-induced (Fig 1, A and B) and C48/80-induced (Fig 1, C and D) dye extravasation. E-G, Hypotension and hypothermia (Fig 1, E) in WT and Hpgds−/− (Fig 1, F) and FVB-background WT and HPGDS-TG (Fig 1, G) mice H, Whole-mount ear sections of WT and Hpgds−/− mice stained with H-PGDS (red) and mast cells (green). Bar = 50 μm. I-M, C48/80-induced (Fig 1, I and J) dye extravasation, PGD2 production (Fig 1, K), hypotension (Fig 1, L), and hypothermia (Fig 1, M) in Hpgdsfl/fl and Mcpt5CreHpgdsfl/fl. Numbers of analyzed animals are shown in parentheses. Data are obtained in 3 independent experiments and are represented as means ± SEMs. *P < .05. i.v., Intravenous. Journal of Allergy and Clinical Immunology  , e9DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 DP receptor mediates the attenuation of anaphylaxis by PGD2, and its signal enhancement inhibits anaphylaxis. A and B, C48/80-induced dye extravasation in the mouse ear. C-E, Effect of DP agonist (BW245C) on C48/80-induced (Fig 2, C and D) dye extravasation on back skin and hypothermia (Fig 2, E). Numbers of analyzed animals are given in parentheses. Data are obtained in 3 independent experiments and are represented as means ± SEMs. *P < .05. i.v., Intravenous. Journal of Allergy and Clinical Immunology  , e9DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig E1 HPGDS deficiency enhances systemic anaphylaxis. A, Antigen/antibody-induced body temperature decrease. B, Hematocrit level 10 minutes after intravenous (i.v.) C48/80 administration. Numbers of analyzed animals are given in parentheses. Data were obtained in 3 independent experiments and are represented as means ± SEMs. *P < .05. Journal of Allergy and Clinical Immunology  , e9DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig E2 C48/80-induced anaphylaxis depends on the histamine H1 receptor in WT and Hpgds−/−. A, Plasma histamine levels 5 minutes after intravenous (i.v.) C48/80 administration. B-D, Effect of the H1 blocker on C48/80-induced (Fig E2, B and C) dye extravasation and hypothermia (Fig E2, D). E-G, Histamine-induced dye extravasation (Fig E2, E and F) and hypothermia (Fig E2, G). Numbers of analyzed animals are given in parentheses. Data were obtained in 2 to 3 independent experiments and are represented as means ± SEMs. *P < .05. Journal of Allergy and Clinical Immunology  , e9DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig E3 C48/80-induced anaphylaxis depends on endothelial nitric oxide synthase (eNOS)–NO in WT and Hpgds−/− mice. A and B, C48/80-induced dye extravasation (Fig E3, A) and hypothermia (Fig E3, B). C-E, Effect of L-NAME on C48/80-induced dye extravasation (Fig E3, C and D) and hypothermia (Fig E3, E). Numbers of analyzed animals are given in parentheses. Data were obtained in 2 to 3 independent experiments and are represented as means ± SEMs. *P < .05. i.v., Intravenous; n.s., not significant. Journal of Allergy and Clinical Immunology  , e9DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig E4 Perivascular mast cells express HPGDS. A and C, Whole-mount immunostaining of an ear section stained with SMA (red) and endothelial cells (blue). Scale bar = 100 μm. B and D, Mean number of mast cells per square millimeter in the ear. E, Whole-mount immunostaining of an ear section stained with HPGDS (red). Mast cells were labeled with avidin-FITC (green). Scale bar = 50 μm. Numbers of analyzed animals are given in parentheses. Data were obtained in 2 to 3 independent experiments. n.s., Not significant. Journal of Allergy and Clinical Immunology  , e9DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Fig E5 PGD2 but not PGD1 inhibits C48/80-induced vascular hyperpermeability. The effect of PGD1 (1 μg) or PGD2 (1 μg) on C48/80-induced dye extravasation on back skin. Numbers of analyzed animals are given in parentheses. Data were obtained in 2 independent experiments and are represented as means ± SEMs. *P < .05. Journal of Allergy and Clinical Immunology  , e9DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

9 Fig E6 DP activation does not induce any decrease in blood pressure. The trace of the blood pressure in vehicle- or DP agonist (BW245C: 1 ng/kg)–treated WT mice is shown. Numbers of analyzed animals are given in parentheses. Data were obtained in 2 independent experiments. Journal of Allergy and Clinical Immunology  , e9DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

10 Fig E7 Generation of Hpgdsfl/fl and Dp−/− mice. A, Strategy for generating the HPGDS targeting vector and HPGDS floxed allele. B, Genotype of Hpgdsfl/fl mice was identified by means of PCR. C, Schematic illustration of DP gene structure and sequences around target loci. Target sequences and PAM and PCR primer loci are indicated by arrows, underlining, and arrowheads, respectively. D, Genotype of Dp−/− mice was identified by means of PCR. Journal of Allergy and Clinical Immunology  , e9DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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