Generation of Potent Cytotoxic T Lymphocytes Against Castration‐Resistant Prostate Cancer Cells by Dendritic Cells Loaded With Dying Allogeneic Prostate Cancer Cells

Abstract

To induce a potent cytotoxic T lymphocyte (CTL) response in dendritic cell (DC)-based immunotherapy against prostate cancer, various tumour antigens should be loaded onto DCs. The aim of this study was to establish a method of immunotherapy for castration-resistant prostate cancer (CRPC) using prostate cancer-specific CTLs generated in vitro by DCs. Monocyte-derived DCs from patients with CRPC were induced to mature using a standard cytokine cocktail (in IL-1b, TNF-a, IL-6 and PGE 2 : standard DCs, sDCs) or using an a-type 1-polarized DC (aDC1) cocktail (in IL-1b, TNF-a, IFN-a, IFN-c and polyinosinic: polycytidylic acid) and loaded with the UVB-irradiated CRPC cell line PC-3. Antigen-loaded DCs were evaluated by morphological and functional assays. The aDC1s significantly increased the expression of several molecules related to DC maturation, regardless of whether the aDC1s were loaded with tumour antigens or not, compared to sDCs. The aDC1s showed a higher production of interleukin-12 both during maturation and after subsequent stimulation with CD40L, which was not significantly affected by loading with tumour antigens, as compared to standard DCs (sDCs). Prostate cancer-specific CTLs against autologous CRPC cells were successfully induced by aDC1s loaded with dying PC-3 cells. Autologous aDC1s loaded with an allogeneic CRPC cell line can generate greater CRPC-specific CTL responses as compared to sDCs and may provide a novel source of DC-based vaccines that can be used for the development of immunotherapy in patients with CRPC.

Generation of potent cytotoxic T lymphocytes against in male patients with non-muscle invasive bladder cancer by dendritic cells loaded with dying T24 bladder cancer cells

“…Furthermore, αDC1s showed a higher production of IL-12, without significant suppression by tumor antigen loading. In addition, potent bladder cancer-specific CTLs against autologous bladder cancer cells from patients were elicited by autologous αDC1s loaded with dying T24 cells, which was consistent withthe results of previous studies on other cancers ( 11 , 16 ).…”

“…On day 6, the iDCs were matured with either conventional cytokine cocktail composed of IL-1ß (25ng/mL, PEPROTECH), TNF-α (50ng/mL, PEPROTECH), IL-6 (1.000units/mL, PEPROTECH), and PGE 2 (106M/L, Sigma-Aldrich, St Louis, MO, USA) to produce sDCs( 8 ), or αDC1-polarizing cytokine cocktail composed of IL-1ß (25ng/mL), TNF-α (50ng/mL), IFN-α (3.000IU/mL, Intron-A-IFN-α-2b, Schering-Plough International, Kenilworth, NJ, USA), IFN-γ (1.000units/mL, Strathmann Biotech GmbH, Hannover, Germany) and poly(I:C) (20μg/mL, Sigma-Aldrich) to produce αDC1s ( 11 ). The DCs were then loaded with the UVB-irradiated T24bladder cancer cell line at a ratio of 2:1 at 2h after the addition of the maturation cytokinesas described previously ( 16 ). The matured DCs loaded with dying T24 tumor cells were harvested on day 8, washed, and analyzed by functional assay.…”

“…To load the tumor cells ontoDCs, T24 cells were irradiated with UVB (30mJ/cm 2 ) (International Light, Newburyport, MA, USA), cultured overnight in RPMI-1640 (Gibco-BRL) supplemented with FBS to induce apoptosis, and then thoroughly washed. The irradiated tumor cells were loaded onto DCs 2h after the addition of maturation cytokines, according to the previous reports ( 16 ). The irradiated dying cells were immediately confirmed using annexin-V and propidium iodide (PI).…”

Antitumor Activity of Lentivirus-mediated Interleukin -12 Gene Modified Dendritic Cells in Human Lung Cancer in Vitro

“…Therefore the lentiviral vector delivery system offers considerable promise for gene therapy on cancer (PicancoCastro et al, 2012;Vannucci et al, 2013). DCs can be generated from multiple cell types using several tissue culture conditions.From peripheral blood, both circulating CD34 hematopoietic progenitor cells and CD14 adherent monocytes have been found to give rise to DCs in presence of granulocyte-macrophage colony-stimulating factor and interleukin-4 (Garg et al, 2013;Hwang et al, 2013). Mature DCs are distinguished by their dendritic oblique morphology and high expression levels of MHC clas1 and class11 as well as CD40/CD80/CD83/CD86 and ICAM-1 (Hansen et al, 2013).…”

Protective effect of Schizandrin B against damage of UVB irradiated skin cells depend on inhibition of inflammatory pathways

“…26,27 Much more IFN-g can be produced by th1 cells and NK cells of immune system with the action effect of IL-18.The cytotoxicity of NK cells was strengthened and the proliferation of th1 cells was activate by IL-18. 28 In recent papers, it assumed that IL-18 can easily bring out malignant skin tumors and was a main product of UVB-irradiation. Effect of Schizandrin B on expression levels of MMP-1 mRNA in FB cells induced by UVB MMP-1, -3and -9 of hearth human epidermis can be induced by UVB-irradiation.…”