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A novel allergen-specific therapy with CD40-silenced B cells and dendritic cells  Motohiko Suzuki, MD, PhD, Makoto Yokota, MD, PhD, Yoshihisa Nakamura,

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Presentation on theme: "A novel allergen-specific therapy with CD40-silenced B cells and dendritic cells  Motohiko Suzuki, MD, PhD, Makoto Yokota, MD, PhD, Yoshihisa Nakamura,"— Presentation transcript:

1 A novel allergen-specific therapy with CD40-silenced B cells and dendritic cells 
Motohiko Suzuki, MD, PhD, Makoto Yokota, MD, PhD, Yoshihisa Nakamura, MD, PhD, Shinya Ozaki, MD, PhD, Shingo Murakami, MD, PhD  Journal of Allergy and Clinical Immunology  Volume 142, Issue 6, Pages e8 (December 2018) DOI: /j.jaci Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Immune modulation by CD40-silenced B cells in vitro. A and B, CD40-silenced B cells were added as inhibitors of OVA-specific (Fig 1, A) or KLH-specific (Fig 1, B) T-cell responses. #P < .05 versus 1:0 group. C, Percentage of Foxp3+ cells in CD4+CD25+ cells after coculture with B cells transfected with reagent alone, control siRNA, or CD40 siRNA. **P < .01 versus no siRNA and control siRNA. D, Foxp3 gene expression determined by using real-time quantitative PCR. **P < .01 versus no siRNA and control siRNA. E and F, CD4+CD25+ cells isolated from coculture with CD40-silenced OVA B cells were added to OVA-specific (Fig 1, E) or KLH-specific (Fig 1, F) T-cell response. **P < .01 versus CD4+CD25+ induced by control siRNA and CD4+CD25−. **P < .01 versus CD4+CD25− induced by CD40 siRNA and CD4+CD25+ induced by control siRNA. Journal of Allergy and Clinical Immunology  , e8DOI: ( /j.jaci ) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Effects of CD40-silenced OVA B cells on established allergic rhinitis. A and B, Numbers of sneezes (Fig 2, A) and rubbing movements (Fig 2, B). C, Number of eosinophils. D, OVA-specific IgE. E, Frequency of CD4+CD25+ T cells in the spleen. F, Foxp3 gene expression in the spleen. G and H, Production of IL-4 (Fig 2, G) and IL-5 (Fig 2, H) by splenocytes stimulated with OVA. I, Percentage of IL-10+ B cells. J, Number of eosinophils in BALF. K, Percentage of CD40+ cells in IL-10+ B cells. B cell×1, 1 × 107 CD40-silenced B cells; B cell×2, 2 × 107 CD40-silenced B cells; B cell+DC, mixture of CD40-silenced B cells and DCs; Control, control B cells; DC×1, 1 × 107 CD40-silenced DCs; DC×2, 2 × 107 CD40-silenced DCs; Normal, no sensitization and treatment. *P < .05 and **P < .01 versus PBS alone and control groups. ##P < .01 versus DC×1. †P < .05 and ††P < .01 versus DC×2. ‡‡P < .01 versus B cell×1. ¶¶P < .01 versus B cell×2. Journal of Allergy and Clinical Immunology  , e8DOI: ( /j.jaci ) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig E1 Modulation by CD40 siRNA. A, CD40 expression analyzed by using FACScan. B cells were transfected with reagent alone (no siRNA), GL2 siRNA (control siRNA), or CD40 siRNA. B, CD40 expression determined by using real-time PCR. C, B-cell proliferation. D, Quantities of IgE released from B cells. **P < .01 versus no siRNA and control siRNA. Journal of Allergy and Clinical Immunology  , e8DOI: ( /j.jaci ) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig E2 Immune modulation by CD40-silenced B cells pulsed with KLH (CD40-silenced KLH B cells) in vitro. CD4+CD25+ or CD4+CD25− cells isolated from coculture with CD40-silenced KLH B cells were added to OVA-specific T-cell response (A) or KLH-specific T cell response (B). *P < .05 and **P < .01 versus CD4+CD25+ induced by control siRNA and CD4+CD25−. Journal of Allergy and Clinical Immunology  , e8DOI: ( /j.jaci ) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig E3 Prevention of OVA-induced allergy by administration of CD40-silenced OVA B cells. Mice that had been treated with intravenous injections of CD40-silenced OVA B cells (CD40), control B cells (Control), or PBS alone (PBS) were sensitized and challenged with OVA, as described in the Methods section. A and B, Numbers of sneezes (Fig E3, A) and nasal rubbing movement (Fig E3, B) were measured after OVA challenge. C, Eosinophilia in the nasal mucosa. D, OVA-specific IgE in sera. E, OVA-specific IgG1 in sera. F and G, Production of IL-10 (Fig E3, F) and IL-35 (Fig E3, G) from splenocytes stimulated with OVA. **P < .01 versus group of PBS alone and control B cells. Journal of Allergy and Clinical Immunology  , e8DOI: ( /j.jaci ) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig E4 CD40-silenced OVA B cells did not prevent KLH-induced allergy. Mice that had been treated with intravenous injections of CD40-silenced OVA B cells (CD40), control B cells (control), or PBS alone (PBS) were sensitized and challenged with KLH, as described in the Methods section. A and B, Numbers of sneezes (Fig E4, A) and nasal rubbing movements (Fig E4, B) were counted after KLH challenge. C, Eosinophilia in the nasal mucosa. D, KLH-specific IgE in sera. Journal of Allergy and Clinical Immunology  , e8DOI: ( /j.jaci ) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Fig E5 Effect of CD40-silenced OVA B cells on established allergic rhinitis caused by KLH. Mice with KLH-induced allergic rhinitis were treated with PBS alone (PBS), control OVA B cells (control), or CD40-silenced OVA B cells (CD40). A and B, Numbers of sneezes (Fig E5, A) and nasal rubbing movements (Fig E5, B) were counted after KLH challenge. C, Eosinophilia in the nasal mucosa. D, KLH-specific IgE in sera. Journal of Allergy and Clinical Immunology  , e8DOI: ( /j.jaci ) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

9 Fig E6 Long-term effect of CD40-silenced OVA B cells. Mice with OVA-induced allergic rhinitis were treated with PBS alone (PBS), control OVA B cells (control), CD40-silenced OVA DCs, CD40-silenced OVA B cells (B cell), or mixture (DC+B) of CD40-silenced OVA DCs and CD40-silenced OVA B cells. Numbers of sneezes (A) and nasal rubbing movements (B) were measured 56 days after treatment. *P < .05 and **P < .01 versus groups of PBS alone and control. #P < .05 ##P < .01 versus DCs. ††P < .01 versus B cells. Journal of Allergy and Clinical Immunology  , e8DOI: ( /j.jaci ) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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