Real time (RT) PCR was performed in triplicate using FAM-labeled Assay-on-Demand reagent sets for IL-10 (Hs00174086_m1) and Foxp3 (Hs00203958_m1). RT-PCR reactions were multiplexed using VIC-labeled 18S primers and probes (Hs99999901_s1) as an endogenous control and analyzed using SDS software version 2.1

(Applied Biosystems), according to the 2-(∆∆Ct) method. Results are presented as mean ± SEM, unless indicated. Data were assessed for normality and equal variation after which the appropriate parametric or nonparametric test was performed (see individual Rapamycin cost figure legends). Differences were considered significant at the 95% confidence level. Correlations were verified with the Pearson’s correlation test or the Spearman’s rank correlation coefficient, as indicated in the figure legend. Z. U. was initially funded by an MRC CASE PhD studentship, held in association with Novartis Institute for Biomedical Research, Horsham, UK. D. R. and Z. U were also supported through funding

by EURO-Thymaide. E. S. C. is funded through an MRC British Thoracic Society/Morriston ABT-199 in vitro Davies Trust Capacity Building PhD studentship. E. X. by a British Lung Foundation Fellowship. C. H. gratefully acknowledges financial support from the Department of Health via the National Institute for Health Research (NIHR) comprehensive Biomedical Research Centre award to Guy’s & St Thomas’ NHS Foundation Trust in partnership with King’s College London and King’s College Decitabine ic50 Hospital NHS Foundation Trust. A. G. is the recipient of a BMA James Trust Fellowship. L. G., J. C., and A. O. G. are funded by MRC, UK. At KCL, we thank C Reinholtz and K Jones, our research nurses. At MRC National Institute for Medical Research we thank: A. Rae, G. Preece, and N. Biboum for assistance in flow cytometry cell sorting; Biological Services Unit

and Xumei Wu for animal husbandry and breeding. We thank Bernard Malissen INSERM-CNRS Universite de la Mediterranee, France and Adrien Kissenpfennig, Queen’s University, UK for their generosity in providing the Foxp3GFP C57BL/6 mice. The authors declare no financial or commercial conflict of interest. Disclaimer: Supplementary materials have been peer-reviewed but not copyedited. Table 1: Characteristics of pediatric asthma patients. For more information see Bush & Saglani, 2010 [47]. Figure 1: Blocking TGF-β signaling diminishes the frequency of Foxp3+ T cells in 1α25VitD3 treated cultures. Figure 2: 1α25VitD3 maintains Foxp3 expression of murine regulatory T cells. Figure 3: Blocking IL-10 signaling promotes the proliferation of Foxp3+ T cells in 1α25VitD3 treated cultures. Figure 4: Purity of peripheral blood Treg and effector T cells isolated by cell sorting. Figure 5: Treg gating strategy in bronchoaveolar lavage fluid. “
“Despite the high prevalence of highly pathogenic H5N1 influenza A viruses in Indonesia, epidemiology information on seasonal human influenza is lacking.