Fig 2: Clusters colored by IFN-I gene expression. (A) Cells colored by IFN-a subtype gene expression. (B) Cells colored by non-IFN-a type I interferon gene expression. Bottom right panel: highlight of clusters 1 and 11 identified by Seurat.

Fig 3: Investigating the sexual dimorphism in the TLR7-, TLR9- and STING-mediated innate cytokine responses with aging. (a) demographic data of the study cohort, age and sex distribution. (b) Schematic summary of study design: Blood samples were obtained from 310 healthy community-dwelling individuals (ages 18–97), samples were separated in two for whole blood stimulation with the indicated PRR ligands, and the other part was used to isolate PBMCs for FACS analysis as indicated. (c, d, e) Levels of cytokine release after whole blood stimulation with the three studied PRR ligands: R848 (c), ODN M362 (d) or cGAMP (e). Concentrations of indicated cytokines were quantified at 24 h in plasma. The results are given in log10 of the concentrations (pg/ml) with background (without ligands) subtracted. (c, d, e, f-h) Sex-differences in IFN-a release in response to R848 (f), ODN M362 (g) and cGAMP (h), before and after the age of 60. Statistical differences between groups were analyzed using the unpaired Student's t-test (c-d) or by a one-way ANOVA test followed by Sidak's multiple comparisons test (f–h).

Fig 4: The female predominance in TLR7-driven production of IFN-a is robustly maintained in elderly subjects above 80. (a–c) From the source groups depicted in Figure 1a (age range >80), the production of IFN-a is shown in response to R848 (a), ODN M362 (b) and cGAMP (c) by sex, female (F), male (M). Statistical differences between groups were analyzed using the unpaired Student's t-test.

Fig 5: Aging-related changes in pDCs and monocytes differ between sexes. (a) Absolute cell counts of pDC, mDC and monocytes per mm3 were calculated from FCM data as described in materials and methods. Absolute counts per mm3 (given in log10) are compared in males and females for individuals under or above 60. (b, c) Correlation between log10 IFN-a release in response to R848 (b) or ODN M362 (c) stimulation and log10 absolute numbers per mm3 of plasmacytoid dendritic cells. Pearson's correlation coefficients “R” and probabilities of correlation were calculated by means of Statview software. (d, e) Residual values (difference between the observed IFN-a concentration and the predicted value) were calculated from the linear regression of IFN-a response as a function of pDC number shown in (b) and (c), respectively. Differences between groups were analyzed using a one-way ANOVA test followed by Sidak's multiple comparisons test (a, d, e).