Toosendanin activates caspase-1 and induces maturation of IL-1[beta] to inhibit type 2 porcine reproductive and respiratory syndrome virus replication via an IFI16-dependent pathway

Porcine reproductive and respiratory syndrome virus (PRRSV) is a prevalent and endemic swine pathogen which causes significant economic losses in the global swine industry. Multiple vaccines have been developed to prevent PRRSV infection. However, they provide limited protection. Moreover, no effective therapeutic drugs are yet available. Therefore, there is an urgent need to develop novel antiviral strategies to prevent PRRSV infection and transmission. Here we report that Toosendanin (TSN), a tetracyclic triterpene found in the bark or fruits of Melia toosendan Sieb. et Zucc., strongly suppressed type 2 PRRSV replication in vitro in Marc-145 cells and ex vivo in primary porcine alveolar macrophages (PAMs) at sub-micromolar concentrations. The results of transcriptomics revealed that TSN up-regulated the expression of IFI16 in Marc-145 cells. Furthermore, we found that IFI16 silencing enhanced the replication of PRRSV in Marc-145 cells and that the anti-PRRSV activity of TSN was dampened by IFI16 silencing, suggesting that the inhibition of TSN against PRRSV replication is IFI16-dependent. In addition, we showed that TSN activated caspase-1 and induced maturation of IL-1[beta] in an IFI16-dependent pathway. To verify the role of IL-1[beta] in PRRSV infection, we analyzed the effect of exogenous rmIL-1[beta] on PRRSV replication, and the results showed that exogenous IL-1[beta] significantly inhibited PRRSV replication in Marc-145 cells and PAMs in a dose-dependent manner. Altogether, our findings indicate that TSN significantly inhibits PRRSV replication at very low concentrations (EC.sub.50: 0.16-0.20 [mu]M) and may provide opportunities for developing novel anti-PRRSV agents. Keywords: Porcine reproductive and respiratory syndrome virus (PRRSV), Toosendanin (TSN), inhibitory activity, gamma-interferon-inducible protein 16 (IFI16), caspase-1; Interleukin-1[beta] (IL-1[beta])