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Allergen-specific sublingual immunotherapy is dose and duration dependent in a murine allergic rhinitis model  Soichi Tofukuji, PhD, Kazufumi Katayama,

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Presentation on theme: "Allergen-specific sublingual immunotherapy is dose and duration dependent in a murine allergic rhinitis model  Soichi Tofukuji, PhD, Kazufumi Katayama,"— Presentation transcript:

1 Allergen-specific sublingual immunotherapy is dose and duration dependent in a murine allergic rhinitis model  Soichi Tofukuji, PhD, Kazufumi Katayama, PhD, Yoshiyuki Nakano, PhD, Satoru Ishida, PhD, Junji Tsuchida, PhD, Minako Tajiri, MSc, Yusuke Shimo, PhD, Hidekazu Tanaka, PhD, Michitaka Shichijo, PhD  Journal of Allergy and Clinical Immunology  Volume 142, Issue 6, Pages e9 (December 2018) DOI: /j.jaci Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 JC SLIT, but not antihistamine or corticosteroid treatment, results in long-lasting dose-dependent effects. A, Experimental design. i.n., Intranasal. B, Sneezing frequency after the third and fourth courses of challenge. Results shown are cumulative data from 3 independent experiments (n = 17-18 per group). C, Representative hematoxylin and eosin–stained nasal sections after the third and fourth courses of challenge. D, JC-specific cytokine production by sLN cells after the fourth course of challenge. Results are presented as means ± SEMs (n = 3). E, Sneezing frequency after the second, third, and fourth challenges (n = 6 for naive, n = 18 for vehicle, and n = 12 for JC-SLIT [S ] high/low groups). **P < .01 compared with the control/vehicle group. †P < .05 and ††P < .01 compared with the high-dose JC SLIT group. Journal of Allergy and Clinical Immunology  , e9DOI: ( /j.jaci ) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 JC SLIT increases numbes of Foxp3+ Treg cells. A, Representative anti-Foxp3–stained nasal sections (turbinate) after the third course of challenge. B, sLN cells were prepared after the third course of challenge and cultured in the presence of Cry j 1 for 24 hours. Foxp3 and Hprt expression was determined by using quantitative RT-PCR. Results are presented as means ± SEMs (n = 3) relative to Hprt expression. Two independent experiments were performed with similar results. C, Percentages of CD103+Foxp3+ CD4 T cells and CD25+Foxp3+ CD4 T cells are presented as means ± SEMs (n = 6). Two independent experiments were performed with similar results. **P < .01 compared with the control/vehicle group. Journal of Allergy and Clinical Immunology  , e9DOI: ( /j.jaci ) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig E1 JC SLIT reduces JC pollen–induced sneezing, nasal scratching, and cytokine production in a dose-dependent manner. A, Experimental design and sampling. BALB/c mice were intranasally (i.n.) sensitized and challenged with JC pollen, as shown. These mice were then treated with SLIT. Control/vehicle groups were treated with PBS. Low-dose and high-dose JC SLIT groups were treated with JC pollen extract equivalent to 1 and 10 μg per dose of Cry j 1, respectively. B, Sneezing and nasal scratching frequencies were determined for 10 minutes immediately after the second or third course of challenge (n = 6). Representative data from 8 (comparing low-dose JC SLIT to vehicle) or 18 (comparing high-dose JC SLIT to vehicle) independent experiments are shown. C, sLN cells were isolated 3 days after the third course of challenge and cultured in the presence of JC pollen extract for 72 hours. Culture supernatants were collected, and cytokine production was measured by using the Cytometric Bead Array (n = 6). *P < .05 and **P < .01 compared with the control/vehicle group. Journal of Allergy and Clinical Immunology  , e9DOI: ( /j.jaci ) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig E2 Effect of JC SLIT on JC pollen–specific cytokine production in sLN cells, superficial cervical lymph node (cLN) cells, and spleen cells. Splenocytes (A) and sLN cells and cLN cells (B) were isolated after the third course of challenge and cultured in the presence of JC pollen extract (equivalent to 10 μg/mL per dose of Cry j 1) for 72 hours. Culture supernatants were collected, and cytokine production was measured by using the Cytometric Bead Array. Results are presented as means ± SEMs (Fig E2, A: n = 6 for naive, n = 9 for vehicle, and n = 17 for high-dose JC-SLIT; Fig E1, B: n = 3). *P < .05 and **P < .01 compared with the control/vehicle group. Journal of Allergy and Clinical Immunology  , e9DOI: ( /j.jaci ) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig E3 Correlation between sneezing frequency and JC pollen–specific cytokine production in sLN cells. Sneezing frequency was determined immediately after the third course of challenge, and then sLN cells were isolated and cultured in the presence of JC pollen extract (equivalent to 10 μg/mL per dose of Cry j 1) for 72 hours. Culture supernatants were collected, and cytokine production was measured by using the Cytometric Bead Array. Results are shown as cumulative data from multiple independent experiments: control/vehicle group (n = 30; n = 6/experiment), low-dose JC SLIT group (n = 18; n = 6/experiment), and high-dose JC SLIT group (n = 30; n = 6/experiment). Correlations between data sets were measured by using the Pearson correlation coefficient. Journal of Allergy and Clinical Immunology  , e9DOI: ( /j.jaci ) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig E4 Effect of JC SLIT on JC pollen–specific immunoglobulin levels. Serum was harvested after the third course of challenge. JC pollen–specific IgE, IgA, IgG1, and IgG2a levels were measured by means of ELISA. Results are presented as means ± SEMs (IgE: n = 6 for all groups; IgA, IgG1, and IgG2a: n = 6 for naive, n = 18 for vehicle, and n = 12 for high-dose JC SLIT). Two independent experiments were performed with similar results. *P < .05 and **P < .01 compared with the control/vehicle group. Journal of Allergy and Clinical Immunology  , e9DOI: ( /j.jaci ) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Fig E5 Effect of administration route on JC pollen–induced sneezing and JC pollen–specific cytokine production in sLN cells. BALB/c mice were intranasally (i.n.) sensitized and challenged with JC pollen. These mice were then administered JC pollen extract through the sublingual (JC-SLIT) or intragastric (JC-IG) routes. Control/vehicle groups were treated with PBS, whereas low-dose JC-SLIT and low-dose JC-IG groups were treated with JC pollen extract equivalent to 1 μg per dose of Cry j 1. High-dose JC-SLIT and high-dose JC-IG groups were treated with JC pollen extract equivalent to 10 μg per dose of Cry j 1. A, Sneezes were counted for 10 minutes immediately after the third course of challenge (n = 11). Results are presented as means ± SEMs. **P < .01 compared with the control/vehicle group. B, sLN node cells were isolated after the third course of challenge and cultured in the presence of JC extract (equivalent to 10 μg/mL per dose of Cry j 1) for 72 hours. Culture supernatants were collected, and cytokine production was measured by using the Cytometric Bead Array. Results are presented as means ± SEMs (n = 6). **P < .01 compared with the control/vehicle group. Journal of Allergy and Clinical Immunology  , e9DOI: ( /j.jaci ) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

9 Fig E6 CD103+Foxp3+ Treg cells are induced by JC SLIT. PBMCs were collected after the third course of challenge and stained with anti-CD4, anti-CD25, anti-CD103, or anti-Foxp3 mAbs. Representative staining profiles of CD103/Foxp3 and CD25/Foxp3 are shown. Journal of Allergy and Clinical Immunology  , e9DOI: ( /j.jaci ) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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