Recombinant Enterovirus A71 2C

Taking berberine (BBR) as the lead, 23 new BBR derivatives were synthesized and examined for their antiviral activities against four different genotype enterovirus 71 (EV71) strains with a cytopathic effect (CPE) assay. Structure-activity relationship (SAR) studies indicated that introduction of a suitable substituent at the 9-position might be beneficial for potency. Among them, compound 2d exhibited most potent activities with IC50 values of 7.12-14.8 M, similar to that of BBR. The effect of 2d was further confirmed in a dose-dependent manner both in RNA and protein level. The mechanism revealed that 2d could inhibit the activation of MEK/ERK signaling pathway. Meanwhile, it could suppress the EV71-induced autophagy by activating AKT and inhibiting the phosphorylation of JNK and PI3KIII proteins. We consider BBR derivatives to be a new family of anti-EV71 agents through targeting host components, with an advantage of broad-spectrum anti-EV71 potency.

Background: Although gamma delta T cells have been reported to be closely related to the immunopathogenesis of some viral infectious diseases, the changes or roles of gamma delta T cells in the development of hand, foot, and mouth disease (HFMD) remain unclear. Methods: Peripheral y8 T cells and their subsets were determined by surface (gamma delta TCR, V31 TCR, V82 TCR, CD45RA, and CD27) or intracellular (IFN-gamma, TNF-alpha, CD107a, and Granzyme B) markers in healthy controls (HCs) and HFMD patients with FACS. The plasma levels of IFN-gamma, TNF-alpha, IL-6, and MCP-1 were measured by ELISA. Differences in gamma delta T cells or their subsets and correlations between gamma delta T cells and inflammation indicators were statistically analyzed. Results: Compared to HCs, HFMD patients showed increased effector gamma delta T and TNF-alpha(+) gamma delta T cells and plasma TNF-alpha levels, especially in severe cases. In addition, significantly increased V delta 1 T and IFN-gamma(+) gamma delta T cells and other plasma inflammatory cytokines were further found in severe patients. Furthermore, EV71 + severe patients showed significantly increased effector and cytokine-producing gamma delta T cells, while the EV71- severe patients displayed significantly greater plasma cytokine levels. The percentage of IFN-gamma(+) gamma delta T or TNF-alpha(+) gamma delta T cells was positively correlated with that of effector y8 T cells. There was a positive correlation between the proportion of V delta 1 T cells and white blood cell (WBC) count or the proportion of IFN-gamma(+) gamma delta T or TNF-alpha(+) gamma delta T cells and neutrophil (N) count, while there was a negative correlation between V delta 2 T cells and WBC or N count. Moreover, the percentages of V delta 1 T and effector gamma delta T cells in the acute phase of disease declined significantly to normal levels during the recovery phase. Conclusions: Increased effector gamma delta T cells with enhanced cytokine production were remarkably observed in severe HFMD patients, which was also associated with clinical inflammation parameters. These data indicated that gamma delta T cells might be involved in inflammatory abnormalities in severe HFMD.