1. CD34+ hemopoietic progenitor cells are potent effectors of allergic inflammation 
Zoulfia Allakhverdi, PhD, Michael R. Comeau, BSc, Dirk E. Smith, MSc, Dean Toy, MSc, Leandra Mfuna Endam, MSc, Martin Desrosiers, MD, Yong-Jun Liu, MD, PhD, Karen J. Howie, MSc, Judah A. Denburg, MD, Gail M. Gauvreau, PhD, Guy Delespesse, MD, PhD 
Journal of Allergy and Clinical Immunology 
Volume 123, Issue 2, Pages e1 (February 2009)
DOI: /j.jaci
Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2. Fig 1
CD34+ hemopoietic progenitor cells express functional receptor for TSLP and IL-33. A, Expression of TSLP and IL-33 receptors at the mRNA (means ± SEMs of 5 experiments) and protein levels. B, Expression of IL-5, IL-13, and GM-CSF mRNA after overnight stimulation (1 representative of 3 experiments). C, IL-5 production after overnight stimulation. Means ± SEMs of 8 experiments. Control IgG1 and control Fc had no effect on the response. D, Expression of CD34 and intracytoplasmic IL-13 after overnight stimulation. Histograms were generated by gating on CD34+ cells. Representative of 3 experiments. ctr, Control. ∗P < .05; ∗∗P < .01; ∗∗∗P < .001.
Journal of Allergy and Clinical Immunology  , e1DOI: ( /j.jaci )
Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3. Fig 2
Activated CD34+ progenitors rapidly release high levels of cytokines and chemokines. Cytokines and chemokines secretion (pg/mL) by freshly isolated neonatal CD34+ cells stimulated with IL-33/TSLP (10 ng/mL each) or IL-1/TNF/TSLP in overnight cultures was measured by ELISA. Means ± SEMs of 6 to 10 experiments. ∗P < .05; ∗∗P < .01; ∗∗∗P < .001.
Journal of Allergy and Clinical Immunology  , e1DOI: ( /j.jaci )
Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4. Fig 3
Cytokines/mediators regulate the response of CD34+ cells to TSLP/IL-33. CD34+ cells were activated overnight with TSLP/IL-33 in the presence or absence of SCF (100 ng/mL) or IL-3 (5 ng/mL) (A); TGF-β and IL-10 (each at 10 ng/mL) (B); IL-1, IL-4, IL-6, IFN-γ, and TNF (each at 10 ng/mL) (C); LTC4, PGE2, and prostaglandin D2 (PGD2; each at 10-6 mol/L) (D); and supernatants were tested for IL-5. Means ± SEMs of 5 to 6 experiments. ∗P < .05; ∗∗P < .01; ∗∗∗P < .001.
Journal of Allergy and Clinical Immunology  , e1DOI: ( /j.jaci )
Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5. Fig 4
CD34+ cells activation by the culture supernatants of primary SAECs and nasal explants. A, CD34+ cells were cultured for 24 hours with the culture supernatants of unstimulated (SNT unstim) or polyI:C-stimulated SAECs (50% vol/vol) in the presence or absence of exogenous IL-1/TNF, neutralizing antibody to TSLP, or isotype control IgG. Means ± SEMs of 3 experiments. B, CD34+ progenitors were cultured with cell-free supernatants (50% vol/vol) of nasal explants from allergic (triangle) and nonallergic (square) patients with chronic rhinitis (Patient SNT) and control subjects (Ctr SNT) in the presence or absence of neutralizing antibody to TSLP. Isotype control antibody had no effect. IL-5 was not detected in the supernatants of nasal explants. ∗P < .05; ∗∗P < .01; ∗∗∗P < .001.
Journal of Allergy and Clinical Immunology  , e1DOI: ( /j.jaci )
Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6. Fig 5
In vivo proinflammatory activity of CD34+ cells. A, The presence of CD34+/IL-5+ and CD34+/IL-13+ cells in the sputum of normal subjects and subjects with allergic asthma at baseline. One representative of 3 from each group is shown. B, The presence of CD34+/IL-5+ and CD34+/IL-13+ cells in the sputum of subjects with allergic asthma after allergen challenge. One representative of 2 experiments is shown. ctr, Control.
Journal of Allergy and Clinical Immunology  , e1DOI: ( /j.jaci )
Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7. IL -5neutralization inhibits eosinophil differentiation but not IL-33/TSLP-induced cytokine production. A, Neonatal CD34+ cells were cultured with SCF in the absence or presence of neutralizing antibody to IL-5 and after 2 weeks of culture were stained for major basic protein (BMK-13; red) to identify eosinophils. One representative of 4 experiments. B, Neonatal CD34+ cells cultured with or without neutralizing goat antibody to IL-5, IL-5R, or normal goat IgG (10 μg/mL each) were stimulated with IL-33/TSLP or IL-1/TNF/TSLP as indicated, and after 1 week of culture, their supernatants were analyzed for IL-5 and IL-13. Means ± SEMs of 4 experiments.
Journal of Allergy and Clinical Immunology  , e1DOI: ( /j.jaci )
Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions