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 3

E2 inhibition reduces the development and IFN signature of TCSle cDCs. Bone marrow-derived cDCs from (a–e) B6 and (a, b, d, e) TCSle female mice were cultured with GM-CSF in standard phenol red IMDM (a–e) and RPMI (c) supplemented with Fulvestrant (Fulvi 1 μM or 100 nM in DMSO) and Tamoxifen (Tam 10 nM or 100 nM in ethanol: EtOH). (a, b, d, e) or with 0 or 0.1 nM E2 (c). On day 7-8, cDCs were harvested and stained with antibodies against CD11c and CD11b and analyzed by flow cytometry. On day 7, cDCs were stimulated with CpG B 1826 (d, e). Six hours post stimulation, cDCs were harvested and total RNA was isolated for qRT-PCR analysis (b, d, e). Mx1 and Cxcl10 RNAs were normalized to the RNA of the housekeeping gene cyclophilin. Standard female B6 control condition was set to 1. Unpaired t-test comparing baseline cDC to SERM-treated groups was used. Mean ± SE values are from 2 independent experiments, using one mouse per strain per experiment (a–e). Brackets indicate significance between controls (including DMSO and EtOH) and SERM-treated samples. , , and .

(a)

(b)

(c)

(d)

(e)