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Fms-like tyrosine kinase 3 ligand increases a lung DC subset with regulatory properties in allergic airway inflammation  Zhifei Shao, MD, Arpita S. Bharadwaj,

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Presentation on theme: "Fms-like tyrosine kinase 3 ligand increases a lung DC subset with regulatory properties in allergic airway inflammation  Zhifei Shao, MD, Arpita S. Bharadwaj,"— Presentation transcript:

1 Fms-like tyrosine kinase 3 ligand increases a lung DC subset with regulatory properties in allergic airway inflammation  Zhifei Shao, MD, Arpita S. Bharadwaj, PhD, Halvor S. McGee, BS, Toluwalope O. Makinde, BS, Devendra K. Agrawal, PhD  Journal of Allergy and Clinical Immunology  Volume 123, Issue 4, Pages e2 (April 2009) DOI: /j.jaci Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Protocol for sensitization, treatment, and adoptive transfer. A, OVA sensitization was followed by methacholine challenge. A total dose of 50 μg of Flt3 ligand (Flt3L) was given to each mouse before the final methacholine challenge to reassess AHR. B, Adoptive transfer of lung DCs. Two DC populations isolated from 3 treatment groups of mice were adoptively transferred to OVA-sensitized and OVA-challenged mice on day 34. i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Dynamics and characteristics of lung DC populations. A, Two lung DC populations: CD11chighCD11blow(squares) and CD11clowD11bhigh (circles; n = 5). Flt3L, Flt3 ligand. B, Expression of cell-surface markers on the 2 lung DC subsets in PBS-treated control mice (n = 5). C and D, Quantitative analyses of the 2 DC subsets. ∗∗∗P < .001 (n = 9-15). Error bars indicate 95% confidence limits. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 T-cell proliferation by 2 DC populations. CD11chighCD11blow DCs isolated from OVA-sensitized and Flt3 ligand (Flt3L)–treated mice showed inability to activate CD4+ T-cell proliferation. ∗∗∗P < .001, n = 5. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 Cytokine secretion patterns in DC/T-cell coculture supernatants and BALF. Levels of IL-12 (A), IFN-γ (B), and IL-10 (C) in DC/T-cell coculture supernatants. D-F, Levels of IL-12 (Fig 4, D), IFN-γ (Fig 4, E), and IL-10 (Fig 4, F) in BALF. ∗∗∗P < .001, ∗∗P < .01, ∗P < .05 (n = 3∼6). Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 5 Adoptive transfer of lung DCs. A, Pulmonary function of sensitized mice after adoptive transfer of DCs from different donors. •••P < .001, CD11chighCD11blow–OVA+Flt3 ligand versus OVA; ♦♦♦P < .001 and ♦♦P < .01, CD11chighCD11blow–OVA+Flt3 ligand versus CD11clowD11bhigh−OVA+ Flt3 ligand; ∗∗∗P < .001 and ∗∗P < .01, CD11chighCD11blow−OVA+Flt3 ligand versus CD11chighCD11blow−OVA (n = 3∼5). The OVA-sensitized and OVA-challenged mice that received adoptively transferred CD11chighCD11blow cells showed an attenuated AHR on day 45. Penh, Enhanced pause. B, Sensitized mice receiving adoptive transfer of CD11chighCD11blow DCs isolated from OVA-sensitized mice treated with Flt3 ligand (OVAAT−OVA) showed decreased airway inflammation and decreased mucus cell hyperplasia (×60 magnification). H&E, Hematoxylin and eosin; PAS, periodic acid–Schiff. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig 6 Expression levels of costimulatory molecules. The expression levels of CD80, CD86 CD40, PDL1, and PDL2 in CD11chighCD11blow and CD11clowD11bhigh DC populations are differentially regulated by Flt3 ligand: A, CD80; B, CD86; C, CD40; D, PDL1; and E, PDL2. ∗∗∗P < .001 and ∗∗P < .01 (n = 6∼12). Error bars indicate 95% confidence limits. MFI, Mean fluorescence intensity. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Flt 3 ligand treatment attenuates AHR to methacholine
Flt 3 ligand treatment attenuates AHR to methacholine. Left, Enhanced pause (Penh) on day 33 (n = 12). ∗∗∗P < .001, OVA day 45 versus PBS day 45. ♦♦♦P < .001, OVA–Flt3 ligand day 45 versus OVA day 45. Right, Lung-specific resistance (RL) on day 45 (n = 5∼8). ∗∗∗P < and ∗∗P < .01, OVA day 45 versus PBS day 45. ♦♦♦P < and ♦♦P < .01, OVA–Flt3 ligand day 45 versus OVA day 45. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

9 Identification of lung plasmacytoid DCs (pDC) by using specific cell-surface markers. A, CD11c bead–purified lung cells in B220 versus CD11b dot plots. G1, B220+CD11b− cells. B, G1 cells in B220 versus Gr-1 dot plot. G2, B220+Cd11b−Gr-1+ cells. C, G2 cells Cd11c versus Cd11b dot plot. G3, Gate for pDCs: B220+Cd11b−Gr-1+ cells. D, CD11c bead–purified lung cells in CD11c versus CD11b dot plot. G3, Gate for pDCs not included in the gates for sorting CD11chighCD11blow and CD11clowD11bhigh lung DCs. E, mPDCA-1 expression in pDCs and CD11c bead–purified lung cells. pDCs have higher levels of mPDCA-1 expression than lung cells. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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