LIF and IL-6 generate M2d TAM-like cells. (A,B,G,H) MΦs, ovarian TAMs, LIF-MΦs, and IL-6-MΦs were stimulated with LPS for 48 hours and IL-10, IL-12, VEGF, and TGFβ were measured by ELISA in the SNs. Results are expressed as means ± SD; n = 6. (C) MMP9, PDGFA, PDGFB, CCL2, CCL5, IL-10, IL-12p35, and IL-23p19 mRNA expression was analyzed by Q-PCR in MΦs, LIF-MΦs, and IL-6-MΦs. ^a indicates that cells were further stimulated for 16 hours with LPS. Results are expressed as the fold increase of mRNA expression in LIF-MΦs or IL-6-MΦs compared with MΦs (mean ± SD; n = 4). (D) MΦs, LIF-MΦs, or IL-6-MΦs were activated for 48 hours with LPS with allogenic CD4⁺ T cells. [³H]-thymidine incorporation was measured at day 4. Results are expressed in cpm (mean ± SD; n = 4). (E,F) LIF-MΦs, IL-6-MΦs, TA-MΦs, and TAMs were activated for 48 hours with LPS and cocultured with CFDA-SE–labeled CD4⁺ T cells stimulated with anti-CD3 mAb plus IL-2 in the presence or absence of 1-MT. At day 5, cells were stained with 7-AAD and annexin-V, and the percentage of living cells was determined by FACS (mean ± SD; n = 4) (E). Proliferation of living cells was evaluated by CFDA-SE dilution measured by FACS. Results are expressed as a percentage of cells in each cycle (mean ± SD; n = 3) (F). (I) PCR analysis of B7-H4 in expression of MΦs, IL-6-MΦs, LIF-MΦs, and TAMs. Result is representative of 1 of 3 experiments. *P < .05.