Fig 2: The FBP–IL-27–COX-2 regulatory axis in decidua is insufficient in patients with unexplained RSA.(A) The mRNA levels of FBP1 and PFK1 between DSCs from normal pregnant women (n = 10) or patients with unexplained RSA (n = 10) were detected by RT-PCR. (B and C) Expression of FBP1 and PFK1 between decidua tissues from normal pregnant women (n = 12) or patients with unexplained RSA (n = 12) were detected by immunohistochemistry. (D) The FBP levels in the blood plasma from nonpregnant (NP) or pregnant (P) normal women or patients with unexplained RSA (n = 10 per group) were detected. (E) The mRNA level of IL-27 between DSCs from normal pregnant women (n = 10) or patients with unexplained RSA (n = 10) was detected by RT-PCR. (F and G) Expression of IL-27 between decidua tissues from normal pregnant women (n = 20) or patients with unexplained RSA (n = 20) was detected by immunohistochemistry. (H and I) The levels of COX-2 (H), CD80, CD86, INF-γ, CD206, CD209, and IRF4 (I) in dMφs from normal pregnant women (n = 12) or patients with unexplained RSA (n = 12) were detected by FCM. Data were presented as means ± SEM and analyzed by t test or one-way ANOVA test. *P < 0.05, **P < 0.01, and ***P < 0.001.

Fig 3: MST1 is downregulated in endometriosis-related peritoneal macrophages. (A, B) Intraperitoneal cells were collected from patients with or without endometriosis and then analyzed by flow cytometry. A higher percentage of CD68+CD86+ macrophages was present in the peritoneal cavity of the control group, while the percentage of CD68+CD163+ macrophages was higher in the peritoneal cavity of endometriosis patients. (C) The IL-10 concentration of human peritoneal fluid supernatant with or without endometriosis. (D, E) Representative western blot of protein expression of MST1, and p38-MAPK. (F) The mRNA level of MST1 and p38-MAPK by RT-qPCR. (G) The location of MST1 and p38-MAPK by immunofluorescence staining. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 (Control group peritoneal macrophages vs. endometriosis group peritoneal macrophages). Values represent the mean ± standard error.

Fig 4: Schematic of proposed mechanism for CDK4/6 inhibitors’ effects on surface immune molecules in KSHV+ cells and EBV+ cells. In addition to direct inhibition of tumor cell proliferation, CDK4/6 inhibitors downregulate DNA methyltransferase 1, which activates both ERVs and certain KSHV/EBV genes. The DNA and RNA viral elements stimulate IFNs, which activate transcription factors including STAT1 and NLRC5. These genes in turn transactivate the expression of ISGs and immune surface molecules including MHC-I, ICAM-1, B7-2, and PD-L1. These surface molecules enable killing of the tumor by binding to receptors on T cells and potentially NK cells too. Because expression of PD-L1 is also enhanced, the results suggest that it may be worth testing CDK4/6 inhibitors with anti-PD1/L1 therapy

Fig 5: CDK4/6 inhibitors increase mRNA expression of MHC-I, ICAM-1, B7-2, and PD-L1 in KSHV+ cells and EBV+ cells. JSC-1 (a), BCBL-1 (b), Akata (c), and Raji (d) cells were treated with 1 μM abemaciclib, or with RPMI medium control for 24 h or 48 h. Total RNA was extracted and analyzed for mRNA expression of MHC-I, ICAM-1, B7-2 and PD-L1 using realtime qPCR. Results shown are mean fold changes relative to untreated cells from 3 experiments. Error bars represent standard deviations from the 3 experiments