Fig. 1

Sex disparity of miR-155 expression. PBMCs were seeded at a concentration of 2 × 10⁶ cells/ml and were cultured in RPMI 1640 media containing 1% L-glutamine, 1% penicillin–streptomycin (all from GE Healthcare Life Sciences HyClone Laboratories, Logan, Utah), 10% heat-inactivated AB+ human serum (in-house), and 100 U/ml recombinant IL-2 (BD Pharmingen™, BD Biosciences). Cells were subsequently subjected to three stimuli: 50 ng/ml recombinant IL-33 (PeproTech, Rocky Hill, New Jersey), a combination of 50 ng/ml IL-33+100 nM of prostaglandin D2 (PGD2; Cayman Chemical, Ann Arbor, Michigan), or 10 µg/ml P(I:C) (Poly(I:C) HMW VacciGrade™ InvivoGen, Toulouse, France) for 22 hours. This study was performed as five separate experiments. Upon the stimulation of cells, a there was no difference between healthy and asthmatic individuals in the numbers of miR-155⁺ILCs. There were sex differences in b the numbers of miR-155⁺ILCs, c and miR-155 expression in ILCs based on the median fluorescence index (MFI) values. Additionally, there were sex differences in the j number of miR-155⁺ILCs expressing ST2 and k the number of miR-155⁺CD4⁺ cells. d Correlation between the numbers of miR-155⁺ILCs and miR-155 expression in all subjects. e Numbers of miR-155⁺ILC2s. Release of f IL-5 protein and g CXCL10 protein in cell-free supernatants. Correlation between miR-155⁺ILC2s and h IL-5 in females and i) CXCL10 in all subjects. All data are expressed as changes from the no stimuli control (∆) for each individual. Female data are shown in red, male data are shown in blue, and data from all subjects are shown in black. *P < 0.05 and **P < 0.01 indicate differences between the sexes; ^ǂP < 0.05, ^ǂǂP < 0.01, and ^ǂǂǂP < 0.005 indicate differences between stimuli; significance is color-coded as previously described. Mann–Whitney tests were used for comparisons between females and males, and the Friedman test followed by Dunn’s multiple-comparison test was used for comparisons between stimuli. Correlations were performed by using Spearman’s rho, with R indicating the Spearman coefficient