PSCs derived Galectin-1 promotes the proliferative activity (S-phase fraction) of CFPAC-1 cells.

<p>(a–c) Quantitative graphical representation of Apoptosis, G1, G2 and S cell population. Points, average of three independent experiments. a, CFPAC-1; b, CFPAC-1 + hNPSC; c, CFPAC-1 + hCaPSC. (d) A bar-graphical representation S-Phase Fraction cells in each group. *<i>p</i><0.01 <i>vs</i>. CFPAC-1, #<i>p</i><0.05 <i>vs</i>. hNPSC+CFPAC-1, ^△<i>p</i>>0.05 <i>vs</i>. hNPSC +β-Lactose.</p

Mechanism of hCaPSCs derived Galectin-1 increased the invasion and metastasis of PDCA.

<p>By secreting the cytokine of TGF-β1, PDAC cells stimulate the activation of quiescent PSCs and promote PSCs to express high level of Galectin-1. Galectin-1 has also been involved in PSCs activation, proliferation and chemokine production, which in turn stimulate the malignant potency of pancreatic cancer cells by increased both the expression of MMP2 and MMP9, and then establish a vicious cycle of mutually reinforcing mechanisms to sustain the activity of the stromal reaction and promotes the invasion and metastasis of PDCA.</p

Galectin-1 intensity of staining and clinicopathologic characteristics of the 18 patients with chronic pancreatitis and 66 patients with pancreatic cancer.

<p>UICC, International Union Against Cancer; PNI, perineural invasion.</p

Univariate and multivariate analysis of conventional prognostic factors and stromal galectin-1 expression in the patients with pancreatic cancer.

<p>Univariate and multivariate analysis of conventional prognostic factors and stromal galectin-1 expression in the patients with pancreatic cancer.</p

Kaplan-Meier survival curve for 66 pancreatic cancers with galectin-1 staining intensity in tumor associated stromal cells.

<p>Kaplan-Meier survival curve for 66 pancreatic cancers with galectin-1 staining intensity in tumor associated stromal cells.</p

Expression of Galectin-1 in primary isolated PSCs and pancreatic cancer cell lines (BxPC-3, SW1990, CFPAC-1 and PANC-1).

<p>(<i>a</i>) Quantitative analyses of the differentiated phenotype of human PSCs and pancreatic cancer cell lines. Higher Galectin-1 mRNA expression was observed in the hCaPSC, and lower Galectin-1 mRNA expression was observed in pancreatic cancer cell lines. *P<0.01 vs. hNPSC, ^#P<0.05 vs. hNPSC. hNPSC, PSC isolated from normal pancreas tissue; hCaPSC, PSC isolated from pancreatic cancer tissue. (<i>b</i>) Typical Western blot indicating stronger band of Galectin-1 in hCaPSC and hNPSC than that in pancreatic cancer cell lines. (<i>c</i>) Strong immunohistochemisty staining of Galectin-1 in hCaPSC. (<i>d</i>) Moderate immunohistochemisty staining of Galectin-1 in hNPSC. Original magnification: ×200.</p

Effect of PSCs derived Galectin-1 on tumor establishment and growth.

<p>(a) CFPAC-1 with or without hCaPSC, hNPSC was implanted subcutaneously on both flanks of nude mice (n = 5), and the mice were sacrificed and xenografts were take out on day 30 after cell implantation. Tumor volume (b) and weight (c) is expressed as the mean±SE. *p<0.01 vs. CFPAC-1, ^#p<0.05 vs. hNPSC+CFPAC-1. Immunohistochemisty staining of Galectin-1 in the xenografts were performed in the group of hCaPSC+CFPAC-1(d), hNPSC+CFPAC-1(e) and CFPAC-1(f). Original magnification: ×200.</p

Endogenous Galectin-1 of PSC upregulated by TGF-β1 and induced MMP2 and MMP9 expression, and promotes the invasion of CFPAC-1.

<p>(A) TGF-β1 regulates Galectin-1 expression in primary cultured hNPSCs. hNPSCs were treated for 24 hours with different concentrations (0.5, 5 and 20 ng/ml) of TGF-β1, which was found induced in a dose-dependent increase of Galectin-1 expression in hNPSCs. (B) TGF-β1 upregulates Galectin-1 expression in the cytoplasm of hNPSCs. Subcellular distribution of Galectin-1 upon TGF-β1 treatment was evaluated by immunohistochemical staining. hNPSCs were treated for 24 and 48 hours with 5 ng/ml TGF-β1 or left untreated. Original magnification: ×200. (C) PSCs derived Galectin-1 increased the expression of MMP-2 and MMP-9 in PCCs. Expression of MMP-2and MMP-9 in PCCs was analyzed by western blot. hCaPSC increased more expression of MMP-2 and MMP-9 in CFPAC-1 than that of hNPSCs, and both of hCaPSC and hNPSCs significantly elevated the expression of both MMP-2and MMP-9 when add 5 ng/ml of TGF-β1 into the supernatant, but β-lactose (50 mM) competitively inhibiting Galectin-1 reduced them. (D)Transwell invasion assay of the pancreatic cancer cell line CFPAC-1. D-a. Invasion of CFPAC-1 in transwell chambers by serum free medium. D-b. Invasion of CFPAC-1 stimulated by hNPSC in transwell chambers. D-c. Invasion of CFPAC-1 stimulated by hCaPSC in transwell chambers. D-d. OD value of each group of invaded CFPAC-1 cells with or without β-Lactose (50 mM). #p<0.05 vs. CFPAC-1, *p<0.05 vs. hNPSC+CFPAC-1, △p<0.05 vs. hNPSC+β-Lactose.</p

Table_1_Interleukin-33 Contributes to the Induction of Th9 Cells and Antitumor Efficacy by Dectin-1-Activated Dendritic Cells.XLS

<p>We recently discovered that dectin-1-activated dendritic cells (DCs) drive potent T helper (Th) 9 cell responses and antitumor immunity. However, the underlying mechanisms need to be further defined. The cytokine microenvironment is critical for Th cell differentiation. Here, we show that dectin-1 activation enhances interleukin (IL)-33 expression in DCs. We found that blocking IL-33/ST2 inhibits dectin-1-activated DC-induced Th9 cell differentiation. More importantly, the addition of IL-33 further promotes Th9 cell priming and antitumor efficacy induced by dectin-1-activated DCs. Mechanistically, in addition to the promotion of Th9 and Th1 cells, dectin-1-activated DCs combined with IL-33 abolish the activity of IL-33 in the induction of regulatory T cells. Furthermore, the combined treatment of dectin-1-activated DCs and IL-33 increases the frequencies of CD4⁺ T cells by fostering their proliferation and inhibiting their exhaustive differentiation. Thus, our results demonstrate the important role of IL-33 in dectin-1-activated DC-induced Th9 cell differentiation and antitumor efficacy, and suggest that the combination of dectin-1-activated DCs and IL-33 may present a new effective modality of DC-based vaccines in tumor immunotherapy.</p

Data_Sheet_1_Interleukin-33 Contributes to the Induction of Th9 Cells and Antitumor Efficacy by Dectin-1-Activated Dendritic Cells.PDF

<p>We recently discovered that dectin-1-activated dendritic cells (DCs) drive potent T helper (Th) 9 cell responses and antitumor immunity. However, the underlying mechanisms need to be further defined. The cytokine microenvironment is critical for Th cell differentiation. Here, we show that dectin-1 activation enhances interleukin (IL)-33 expression in DCs. We found that blocking IL-33/ST2 inhibits dectin-1-activated DC-induced Th9 cell differentiation. More importantly, the addition of IL-33 further promotes Th9 cell priming and antitumor efficacy induced by dectin-1-activated DCs. Mechanistically, in addition to the promotion of Th9 and Th1 cells, dectin-1-activated DCs combined with IL-33 abolish the activity of IL-33 in the induction of regulatory T cells. Furthermore, the combined treatment of dectin-1-activated DCs and IL-33 increases the frequencies of CD4⁺ T cells by fostering their proliferation and inhibiting their exhaustive differentiation. Thus, our results demonstrate the important role of IL-33 in dectin-1-activated DC-induced Th9 cell differentiation and antitumor efficacy, and suggest that the combination of dectin-1-activated DCs and IL-33 may present a new effective modality of DC-based vaccines in tumor immunotherapy.</p