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Restrictive IL-10 induction by an innocuous parainfluenza virus vector ameliorates nasal allergy  Keiichi Yamanaka, MD, Takehisa Nakanishi, MD, Kana Isono,

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Presentation on theme: "Restrictive IL-10 induction by an innocuous parainfluenza virus vector ameliorates nasal allergy  Keiichi Yamanaka, MD, Takehisa Nakanishi, MD, Kana Isono,"— Presentation transcript:

1 Restrictive IL-10 induction by an innocuous parainfluenza virus vector ameliorates nasal allergy 
Keiichi Yamanaka, MD, Takehisa Nakanishi, MD, Kana Isono, MS, Chisami Hasegawa, MD, Hiroyasu Inada, MD, Kento Mizutani, MD, Yoshiaki Matsushima, MD, Karin Okada, MD, Tomotaka Mabuchi, MD, Makoto Kondo, MD, Akisa Yamagiwa, MD, Masato Kakeda, MD, Koji Habe, MD, Tetsuya Nosaka, MD, Esteban C. Gabazza, MD, Hidetoshi Yamazaki, DDS, Hitoshi Mizutani, MD, Mitsuo Kawano, PhD  Journal of Allergy and Clinical Immunology  Volume 139, Issue 2, Pages e7 (February 2017) DOI: /j.jaci Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Visualization of IL-10–producing cells in a mouse model of nasal pollinosis. A, JCP-challenged mice showed increased number of IL-10–producing CD19+ B cells and CD11b+ myeloid cells, and less number of CD4+ T cells and CD8+ T cells. Green: IL-10-GFP; red: CD4+, CD8+, CD19+, or CD11b+; yellow: both IL-10 and CD4/CD8/CD19/CD11b. Representative microphotographs of H&E-stained sample, CD4, CD8, CD19, and CD11b costaining are shown. B, Flow cytometric analysis was performed to count the source of IL-10 in JCP-challenged IL-10 tiger mice. Nasal tissue homogenates were obtained from 5 mice and mononuclear cells were purified through nylon mesh. Cell surface staining was performed with CD4/CD8/CD19/CD11b antibodies. GFP+IL-10–producing mononuclear cells were gated, and then CD4/CD8/CD19/CD11b-positive cells were measured. CD19+ and CD11b+ cells are more as IL-10–producing cells compared with CD4 and CD8 cells. GFP, Green fluorescence protein; H&E, hematoxylin and eosin stain. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Anti-inflammatory effects of rhPIV2/IL-10. A, Schematic diagram of the recombinant hPIV2/IL-10 construct. B, The number of sneezing episodes was significantly suppressed in rhPIV2/IL-10–treated mice (Wilcoxon signed-rank test, n = 8, *P < .05). C, Representative microphotographs of H&E, Toluidine Blue, Giemsa, and other immune staining are also shown. H&E, Hematoxylin and eosin stain; IL-10, rhPIV2/IL-10–treated pollinosis mice; PBS, PBS-treated pollinosis mice; PIV, rhPIV2-treated pollinosis mice. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 2 Anti-inflammatory effects of rhPIV2/IL-10. A, Schematic diagram of the recombinant hPIV2/IL-10 construct. B, The number of sneezing episodes was significantly suppressed in rhPIV2/IL-10–treated mice (Wilcoxon signed-rank test, n = 8, *P < .05). C, Representative microphotographs of H&E, Toluidine Blue, Giemsa, and other immune staining are also shown. H&E, Hematoxylin and eosin stain; IL-10, rhPIV2/IL-10–treated pollinosis mice; PBS, PBS-treated pollinosis mice; PIV, rhPIV2-treated pollinosis mice. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig E1 The number of IL-10–producing cells in a mouse model of nasal pollinosis. JCP-challenged mice showed increased number of IL-10–producing CD19+ B cells and CD11b+ myeloid cells, and less number of CD4+ T cells and CD8+ T cells on day 56 compared with the sections from PBS-treated IL-10 tiger mice (Wilcoxon signed-rank test). A P value of less than .05 was considered statistically significant (n = 6; **P < .001, *P < .05). Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig E2 The suppression of enhanced inflammatory response by rhPIV2/IL-10 treatment. The number of mononuclear cells, neutrophils, and eosinophils was increased in the PBS- or rhPIV2-treated mice, but these infiltrated cell numbers were significantly decreased in rhPIV2/IL-10–treated mice (Wilcoxon signed-rank test; n = 8, **P < .001, *P < .05). Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig E3 A and B, The detection of PIV2. Immunostaining was performed using anti-PIV2 antibody.E1 PIV2 was detected in rhPIV2/IL-10–treated mice, suggesting the infection of rhPIV2/IL-10-vector. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Fig E4 The cytokine concentration in nasal secretion. The concentration of the inflammatory cytokines including IL-1β, IL-2, IL-4, IL-5, IL-10, and TNF-α in the nasal secretion was dramatically suppressed in rhPIV2/IL-10–treated mice (Wilcoxon signed-rank test; n = 8, **P < .001, *P < .05). Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

9 Fig E5 Cytokine production by lymphocytes from cervical lymph node (LN) and spleen. A, IL-4 and IL-13 production was measured by flow cytometry. The percentage of cytokine-producing lymphocytes was similar between rhPIV2 and rhPIV2/IL-10 treatment groups. B, Similarly IFN-γ– and TNF-α–producing lymphocytes were not different between rhPIV2 and rhPIV2/IL-10 application. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

10 Fig E6 Plasma IgE and cytokine concentration, and histamine-releasing assay. A, Plasma IgE level was increased in the JCP group and the levels were unchanged with rhPIV2 or rhPIV2/IL-10 application. B, Plasma TH2 cytokine IL-4, IL-5, and IL-6 concentration was measured with flowcytomix. IL-4 was undetected in all samples. IL-5 and IL-6 were detected, but unchanged in all JCP applied groups even treated with rhPIV2 and rhPIV2/IL-10. C, Histamine-releasing assay was performed using plasma from 4 groups. A total of 3 × 104 mouse mast cells (ATCC, CRL-8306) were cultured in 150 μL of culture medium (ATCC ) supplemented with 20 IU of JCP and 50 μL of plasma samples, and cultured for 8 hours. Histamine level was measured by ELISA (SPI-BIO A05890) and was increased in the plasma from JCP mice. The levels were unchanged even using the plasma from rhPIV2- and rhPIV2/IL-10–applied mice. Statistical analysis was performed using the Kruskal-Wallis nonparametric ANOVA test with post hoc analysis using Dunn's multiple comparison test. A P value of less than .05 was considered statistically significant (n = 6; *P < .05). Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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