This study indicates that the intrahepatic immune responses involved in the clearance of HCV are different between animals in which the immune system has been primed by vaccination and rechallenged animals where the immune system has been primed by natural infection with HCV. Low density arrays may be a useful method to select immune response markers to predict the outcome of HCV infection or the success of a vaccine. Disclosures: Esther Chang – Consulting: SynerGene Therapeutics, Inc. Kathleen F. Pirollo – Grant/Research Support: SynerGene Therapeutics, Inc Stephen Feinstone – Independent Contractor: Dynavax The following people have nothing to disclose: Hongying Duan, Iryna Zubkova, Youkyung Choi, Frances
Wells, Kris Krawczynski, Robert Lanford, Marian E. Major [Background] It has been reported that MDSC and Tregs were major suppressors of the immune response http://www.selleckchem.com/products/epz015666.html against Hepatocel-lular carcinoma (HCC). Sorafenib, an oral multi-kinase inhibitor, has been approved for the treatment of HCC. Sorafenib could inhibit the MAPK and VEGF signaling. VEGF signaling might affect MDSC development as well as angio-genesis. [Aim] The aim of this study is to analyze whether sorafenib could suppress MDSC and Tregs development in HCC patients ex vivo and in vitro. [Methods] ex vivo analysis: Thirty-five HCC patients who received DZNeP molecular weight sorafenib were enrolled in this study. Sorafenib exhibits inter-individual
pharmacokinetic variability based medchemexpress on the activity of CYP3A4. Therefore, we quantitated the sorafenib and sorafenib N-oxide in serum by an optimized HPLC-UV led method. The linear range of detection was 0.03–30 μg/ml. Peripheral blood mononuclear cells (PBMCs) were used for the analysis of MDSCs, Tregs and Th1. PBMCs were stained with CD3, CD4, CD25, CD127, CCR5, CXCR3, CD11 b, CD14, CD16, CD33, PD-L1, and HLA-DR antibody and analyzed by FACS canto-II. IL10 or IFN-γ secreting cells were analyzed by cytokine secreting assay. The mRNA expression of PBMCs was analyzed by deep sequence analysis (Transcriptome analysis) and realtime-PCR analysis (GM-CSF, IFN-γ,IL10,
TGF-β1, arginase 1, iNOS, PD-L1). in vitro analysis: Isolated PBMCs were used to analyze the induction of MDSC and Tregs by the soluble factor induced from various hepatoma cell lines (Hep3B, Li3, PLC etc.) in a 0.4μm pore tran-swell system. NOG mice were used for the transplantation of HCC with MDSC. [Results] ex-vivo: The frequency of MDSC in HCC patients was significantly higher than those in healthy subjects. The frequencies of PD-L1 high MDSCs and Tregs were significantly decreased after 8 weeks sorafenib treatment (p<0.01). On the other hand, the frequency of Th1 cells and the ability of IFN-γ secretion in T cells were significantly increased after 8 weeks sorafenib treatment (p<0.01). The expression of GM-CSF mRNA was significantly decreased after 8 weeks sorafenib treatment (p<0.05).