Conventional DCs from Male and Female Lupus-Prone B6.NZM Sle1/Sle2/Sle3 Mice Express an IFN Signature and Have a Higher Immunometabolism That Are Enhanced by Estrogen

Figure 4

The IFN signature of TCSle cDCs is not due to a differential heterogeneity of the GM-CSF cDCs. Bone marrow precursors from B6 or TCSle female and male mice were cultured with GM-CSF in standard phenol red/media conditions or media depleted of phenol red and devoid of steroids (charcoal-treated FBS: 0 E2) and supplemented with 0.03 nM, 0.1 nM, or 50 nM 17-β-estradiol (E2). Standard conditions were also supplemented with 1 μM Fulvestrant. On day 8, cDCs were harvested and stained with fixable viability dye and antibodies against CD11c, CD11b, MHCII, and CD86. Samples were gated on singlets, live cells, CD11c⁺ MHCII⁺ (a, left), CD11b + versus MHCII^High or MHCII^Int (a, center), and CD86 (a, right). Gray histograms represent the MHCII^High population, while green histograms represent the MHCII^Int population and the black line represents an unstained population. (a) One representative plot of a B6 female culture is shown. Graphs of %MHCII^High (b) or %MHCII^Int (c) from the CD11c⁺ gate from B6 (black closed symbols) or TCSle (gray open symbols) female (circle) and male (triangle) mice. Graphs of %CD86 positive on MHCII^High (d) or MHCII^Int (e) populations. Mean + SE values are from 3 biological replicates, using one mouse of each strain and sex per experiment. Two-way ANOVA analysis with Tukey multiple comparisons was used to compare differences between B6 and TCSle, and results are shown in a box surrounding the symbol Δ for significance between B6 and TCSle males or the symbol O for significance between B6 and TCSle females. Δ and O represent . ΔΔ and OO represent .

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