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Matthew J. Loza, PhD, Susan Foster, PhD, Stephen P

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1 Interactive effects of steroids and β-agonists on accumulation of type 2 T cells 
Matthew J. Loza, PhD, Susan Foster, PhD, Stephen P. Peters, MD, PhD, Raymond B. Penn, PhD  Journal of Allergy and Clinical Immunology  Volume 121, Issue 3, Pages 750.e1-755.e3 (March 2008) DOI: /j.jaci Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Effects of β-agonists on IL-13+ T-cell accumulation. PBLs from asthmatic subjects were cultured for 6 days with IL-2 and anti-IL-12 without (control condition) or with added β-agonist: A, 1 μmol/L isoproterenol; B, indicated concentrations of formoterol or albuterol (x-axis). In Fig 1, A, the percentage and number of IL-13+ T cells after culture with isoproterenol (x-axis) and in Fig 1, B, number of IL-13+ T cells after culture with added formoterol/albuterol is expressed as a percentage of the value observed in the IL-2–stimulated control (ie, no β-agonist) condition (y-axis) for each subject tested. ∗P < .05, ∗∗P < .01, and ∗∗∗P < .001 for the difference in the median (Fig 1, A) or GM (Fig 1, B) from the IL-2–stimulated control condition. †P < .05 for the difference between 6 and 100 nmol/L formoterol. ‡P < .01/.05 for the difference in GM between 700/1400 nmol/L and 10,000 nmol/L albuterol. Journal of Allergy and Clinical Immunology  , 750.e1-755.e3DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Effects of β-agonists in the absence of IL-12 neutralization. PBLs were analyzed for IL-13 production before (day 0) and after 6 days of culture with IL-2 without (−) or with (+) added anti–IL-12 mAb and vehicle, 3 nmol/L formoterol, or 700 nmol/L albuterol. Results are plotted as a percentage of day 0 values. Numbers above plots are the percentage increase in IL-13+ T cells relative to the respective vehicle condition (without or with added anti-IL-12 mAb). ∗P < .01. †P < .001. Journal of Allergy and Clinical Immunology  , 750.e1-755.e3DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 Effects of β-agonists on CD4+ and CD8+ T-cell subsets. PBLs were analyzed for IL-13 production before (day 0) and after 6 days of culture with IL-2 plus anti–IL-12 plus vehicle, 3 nmol/L formoterol, or 700 nmol/L albuterol. Results are plotted as a percentage of day 0 values for CD4+ and CD8+ T-cell subsets. Numbers above plots are the percentage increase in IL-13+ T cells relative to the respective vehicle condition. ∗P < .01 and †P < .05. Journal of Allergy and Clinical Immunology  , 750.e1-755.e3DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 Interactive effects of β-agonist and budesonide. Relative number of IL-13+ T cells after culture of PBLs from asthmatic subjects with IL-2, anti–IL-12, and the indicated concentrations of budesonide and formoterol (A and C) or albuterol (B). ∗P < .05, †P < .01, and ‡P < .001 for the difference in mean from the indicated β-agonist condition without budesonide (n = 10 asthmatic subjects [Fig 4, A and B]; n = 5 subjects in an independent asthmatic population [Fig 4, C]). Journal of Allergy and Clinical Immunology  , 750.e1-755.e3DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 5 Effects of β-agonists and budesonide on total and IFN-γ+ T cells. The relative number of total (left) and IFN-γ+(right) T cells after culture of PBLs from asthmatic subjects with IL-2, anti–IL-12, and the indicated concentrations of budesonide and formoterol (A) or albuterol (B) is shown. ∗P < .05, ∗∗P < .01, and ∗∗∗P < .001 for the difference in mean from the indicated β-agonist condition without budesonide (n = 9-10 asthma subjects). Journal of Allergy and Clinical Immunology  , 750.e1-755.e3DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Effects of β-agonists on IL-13+ T-cell accumulation
Effects of β-agonists on IL-13+ T-cell accumulation. Absolute numbers of IL-13+ T cells before (day 0) and after culture of 1 × 106 PBLs from asthmatic subjects with IL-2, anti-IL-12 alone (control condition [Ctr]), or the indicated concentrations of formoterol or albuterol. ∗P < .05, ∗∗P < .01, and ∗∗∗P < .001 for the difference in GM from IL-2–stimulated control condition. †P <0.05 for the difference between 6 and 100 nmol/L formoterol. ‡P < .01/.05 for the difference in GM between 700/1400 nmol/L and 10,000 nmol/L albuterol. Journal of Allergy and Clinical Immunology  , 750.e1-755.e3DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Effects of β-agonist and budesonide on IL-13+ T cells from control subjects. Relative numbers of IL-13+ T cells after culture of PBLs from control subjects with IL-2, anti–IL-12, and the indicated concentrations of budesonide and formoterol (A) or albuterol (B) are shown. Journal of Allergy and Clinical Immunology  , 750.e1-755.e3DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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