Antiviral activity of shikonin ester derivative PMM-034 against enterovirus 71 in vitro

Human enterovirus 71 (EV71) is the major causative agent of hand, foot, and mouth disease (HFMD), particularly in infants and children below 4 years of age. Shikonin is a bioactive compound with anti-inflammatory, antiviral, and antibacterial activities derived from the roots of the Chinese medicinal herb Lithospermum erythrorhizon. This study aimed to examine the antiviral activity of PMM-034, a shikonin ester derivative, against EV71 in rhabdomyosarcoma (RD) cells. Cytotoxicity of PMM-034 on RD cells was determined using WST-1 assay. Dose- and time-dependent effects of PMM-034 on EV71 replication in RD cells were determined using plaque reduction assay. mRNA expression levels of EV71/VP1 and pro-inflammatory cytokines (IL-1β, IL-6, IL-8, and TNF-α) were determined by real-time RT-PCR, and EV71/VP1 and phospho-p65 protein expressions were determined by western blot analysis. PMM-034 exhibited only weak cytotoxicity against RD cells. However, PMM-034 exhibited significant antiviral activity against EV71 in RD cells with 50% inhibitory concentration of 2.31 μg/mL. The VP1 mRNA and protein levels were significantly reduced in cells treated with PMM-034. Furthermore, relative mRNA expression levels of IL-1β, IL-6, IL-8, and TNF-α significantly decreased in the cells treated with PMM-034, while the phospho-p65 protein expression was also significantly lower in the treated cells. These results indicated that PMM-034 suppressed the expressions of pro-inflammatory cytokines in RD cells, exhibiting antiviral activity against EV71, as evidenced by the reduced VP1 mRNA and protein levels in PMM-034-treated cells. Thus, PMM-034 is a promising candidate for further development as an EV71 inhibitor.

Effect of dendritic cell vaccine therapy on lymphocyte subpopulation in refractory primary brain tumor.

BACKGROUND: Dendritic cell (DC)‑based immunotherapy has the potential to induce an antitumor response within the immunologically privileged brain. AIMS: The aim of this study was to evaluate the short‑term effect of DC vaccine therapy on lymphocyte subsets in patients with refractory primary brain tumor. MATERIALS AND METHODS: Eighteen cases with refractory primary brain tumor who refused any treatment against tumor within 6 months of the therapy, were referred to one medicine center, from January 2011 to October 2012. All patients received 1 × 107 tumor lysate–pulsed DC vaccinations both intradermal injection and intravenous infusion 3 times/week. RESULTS: There were increases of lymphocytes CD8+ (P = 0.002) and CD56+ (P = 4.207E‑10), but no change of lymphocytes CD3+ (P = 0.651). Six patients were positive response of delayed‑type hypersensitivity. There were improving of appetite in 14 cases and increasing of physical strength 17 cases. CONCLUSIONS: DC vaccine has the potential for inducing an immune cytotoxic effect directed toward tumor cells.

Dendritic cell vaccine treatment of advanced de novo colorectal cancer in renal transplant patients.

Objective: The clinical outcome, especial the immunologic responses to cancer and graft, of dendritic cell (DC) vaccine in the treatment of advanced de novo colorectal cancer (CRC) in renal transplant patients was investigated in this study. Materials and Methods: 7 patients were received 1 cycle tumor lysate pulsed autologous DC vaccine. The positive cell-mediated cytotoxicity responses to DC vaccine against CRC cell in two out of 7 patients were seen by delayed type hypersensitivity (DTH) test. The positive cell-mediated cytotoxicity responses to DC vaccine against normal kidney cell in all 7 patients were not seen by DTH tests and no notable change of renal function during and after vaccination. Conclusions: DC vaccine has emerged as a promising new strategy in the treatment of advanced de novo CRC in renal transplant patients and DC vaccines have become an attractive therapeutic option, developing immune responses specific against CRC cell, achieving clinical efficacy without graft failure.