Fig 1: NKp30-B7H6 interaction and tumour-derived PGD2 favour IL-13 secretion. a Cytokine concentrations in serum samples of HD and APL patients (APL) (n = 11). b Representative examples of flow cytometry analysis of cytokine production by ILC in HD and APL patients (APL). c Frequencies of cytokine producing ILC in HD and APL patients (APL), upon co-culture with APL cell lines for 48 h (six independent experiments). d Representative example of flow cytometry analysis of IL-13-positive cells upon co-culture with autologous APL blasts. e Frequencies of IL-13+ ILC2, NKT and Th2 cells (n = 6). f Expression of B7H6 and g frequency of B7H6-positive cells in APL cell lines (NB4, HL60) and a control cell line (721.221, lymhphoblastoid cell line) (six independent experiments). h Representative example of flow cytometry analysis of the expression of B7H6 and i frequency of B7H6-positive cells in APL blasts in bone marrow (APL BM), in APL blasts in peripheral blood (APL PB), (n = 11) or in a control cell line (721.221, n = 6). j Representative example of flow cytometry analysis of the expression of NKp30 on ILC2 in peripheral blood of healthy donors (HD PB), in APL BM or in APL PB. k Relative frequencies of NKp30 expressing ILC2 in HD PB, APL BM or APL PB (n = 11). l Representative example of flow cytometry analysis of IL-13 produced by ILC2 co-cultured with the APL cell line NB4 in the presence of an anti-NKp30 blocking antibody (aNKp30), Ig control or medium. m Frequency of ILC2 IL-13+ in medium (n = 9), in the presence of an aNKp30 or an Ig control (IgM Ctrl) (n = 6, three independent experiments). n Quantification of IL-33, IL-25, TSLP and PGD2 in the sera of HD and APL patients (APL) (n = 11). o Quantification of PGD2 in supernatants of leukaemic AML (KG1, THP1) and APL cell lines (NB4, HL60), in the absence or presence of arachidonic acid (ArAc) (1 experiment). Error bars are s.e.m. Statistical analysis was performed using Mann-Whitney test (a–c, n) and Kruskal-Wallis test (d–i, m) and ANOVA test (k)
Fig 2: ILC2 are significantly increased in prostate cancer. a Quantification of PGD2 in supernatants of different human cancer cell lines (n = 2). b Representative example of flow cytometry analysis of expression of B7H6 in prostate cancer cell lines DU145 and PC3, or in a control cell line (721.221). c Relative frequencies of the ILC2 subset among total ILCs in healthy donors (HD) and prostate cancer patients (Prostate) (n = 21). d Relative frequencies of NKp30 expressing ILC2 in peripheral blood of HD and Prostate (n = 21). e Frequency of M-MDSC in peripheral blood of HD and Prostate (n = 21). f Relative frequencies of the ILC2 subset among total ILCs in healthy donors (HD, n = 21) and prostate cancer patients at stage (Gleason score) 6 (n = 8), 7 (n = 8), or 9/10 (n = 5) or in patients with benign prostate hypertrophy (BPH, n = 5). g Representative examples of flow cytometry analysis of innate lymphoid cell subsets in blood of FVB female mice (blood CTR, n = 5) and in blood (n = 29), spleen (n = 29) and prostate (n = 19) of TRAMP mice. h Relative frequencies of the ILC2 subset among total ILCs in blood of FVB female mice (blood CTR) and in blood, spleen and prostate of TRAMP mice. i Correlation of ILC2 and M-MDSCs in prostate tumours of TRAMP mice (n = 19) (three independent experiments). Error bars are s.e.m. Statistical analysis was performed using t test (c–e, h) and Kruskal-Wallis test (f)
Fig 3: ATRA reverses the PGD2-ILC2-IL-13-M-MDSC immunosuppressive axis. a Schematic representation of the hypothetical immunosuppressive chain established in APL patients at diagnosis. b Comparison of serum concentrations of PGD2 (APL = 11; Rem = 9), c frequencies of ILC2 NKp30+ (APL = 11, APL Rem = 9), d frequencies of total ILC2 (APL = 22; APL Rem n = 9), e serum concentrations of IL-13 (APL n = 11; APL Rem n = 9) and f frequencies of M-MDSC (APL n = 22; APL Rem = 9) in peripheral blood of APL patients at diagnosis (APL) or in remission after ATRA treatment (APL Rem). Dashed lines represent mean values of all the parameters in HD. g Schematic representation of APL establishment and ATRA treatment schedule in FVB/NJ mice upon injection of APL splenocytes (three independent experiments), as assessed by quantification of the normal composition of the bone marrow (Gr1+CD11b+ cells) h, as previously described27. i Survival curves of untreated (APL, n = 13) and ATRA-treated (ATRA) APL mice (n = 6). j Quantification of PGD2 concentrations (CTR n = 15, APL n = 13, ATRA day 14 n = 9, ATRA day 25 n = 6), k ILC2 frequencies and l M-MDSCs frequencies in control FVB/NJ mice (CTR), APL mice before (APL) and after ATRA treatment (ATRA day14; ATRA day25) (CTR n = 12, APL n = 10, ATRA day 14 n = 9, ATRA day 25 n = 6). m Schematic representation of APL establishment and ATRA treatment schedule in HIS mice upon injection of an APL cell line (two independent experiments). n Survival curves of untreated (APL, n = 5) and ATRA-treated (ATRA, n = 3) APL HIS mice. o Quantification of PGD2 concentrations (CTR n = 8, APL n = 8, ATRA n = 5), p ILC2 frequencies (CTR n = 11, APL n = 13, ATRA n = 9) and q M-MDSCs frequencies (CTR n = 8, APL n = 8, ATRA n = 5) in APL HIS mice before (APL) and after ATRA treatment at day 25 (ATRA) (up to two independent experiments). Error bars are s.e.m. Statistical analysis was performed using Mann-Whitney test (b–f), Kruskal-Wallis test (h, j–l, o–q) and Log-rank test (Mantel-Cox) (i, n)
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