SUMMARY During anti-viral defense, interferon (IFN) signaling triggers nuclear transport of tyrosine phosphorylated STAT1 (PY-STAT1), which occurs via a subset of karyopherin alpha (KPNA) nuclear transporters. Many viruses, including Ebola virus, actively antagonize STAT1 signaling to counteract the antiviral effects of IFN. Ebola virus VP24 protein (eVP24) binds KPNA to inhibit PY-STAT1 nuclear transport and render cells refractory to IFNs. We describe the structure of human KPNA5 C-terminus in complex with eVP24. In the complex, eVP24 recognizes a unique non-classical nuclear localization signal (NLS) binding site on KPNA5 that is necessary for efficient PY-STAT1 nuclear transport. eVP24 binds KPNA5 with very high affinity to effectively compete with and inhibit PY-STAT1 nuclear transport. In contrast, eVP24 binding does not affect the transport of classical NLS cargo. Thus, eVP24 counters cell-intrinsic innate immunity by selectively targeting PY-STAT1 nuclear import while leaving the transport of other cargo that maybe required for viral replication unaffected.

Hepatitis E virus (HEV) is a zoonotic pathogen of which pigs are reservoirs. To determine the presence of HEV RNA in commercial pig livers sold in local grocery stores in the USA, 127 packages of commercial pig liver were purchased and tested by a universal RT-PCR assay capable of detecting all four known HEV genotypes. Among the 127 livers tested, 14 were positive for HEV RNA. Sequence and phylogenetic analyses revealed that the 14 isolates all belonged to genotype 3. An animal study was subsequently conducted in pigs to determine whether the PCR-positive pig livers still contained infectious virus. The results showed that pigs inoculated with two of the three PCR-positive pig-liver homogenates became infected, as evidenced by the detection of faecal virus shedding, viraemia and seroconversion. The data demonstrated that commercial pig livers sold in grocery stores are contaminated by HEV and that the contaminating virus remains infectious, thus raising a public-health concern for food-borne HEV infection. The GenBank/EMBL/DDBJ accession numbers for the sequences determined in this study are EF107626-EF107639.

Hepatitis E virus (HEV) is a zoonotic pathogen and pigs are a known reservoir. Recently we showed that approximately 11% of commercial pig livers sold in local U.S. grocery stores for food consumptions are contaminated by infectious HEV. In this study, a swine bioassay was used to determine if the infectious HEV in contaminated commercial pig livers could be inactivated by traditional cooking methods. Group 1 pigs (n=5) were each inoculated intravenously (i.v.) with a HEV-negative liver homogenate as negative controls, group 2 pigs (n=5) were each inoculated i.v. with a pool of two HEV-positive pig liver homogenates as positive controls, groups 3, 4 and 5 pigs (n=5, each group) were each inoculated i.v. with a pool of homogenates of two HEV-positive livers incubated at 56 degrees C for 1 h, stir-fried at 191 degrees C (internal temperature of 71 degrees C) for 5 min or boiled in water for 5 min, respectively. As expected, the group 2 positive control pigs all became infected whereas the group 1 negative control pigs remained negative. Four of the five pigs inoculated with HEV-positive liver homogenates incubated at 56 degrees C for 1 h also became infected. However, pigs in groups 4 and 5 did not become infected. The results indicated that HEV in contaminated commercial pig livers can be effectively inactivated if cooked properly, although incubation at 56 degrees C for 1 h cannot inactivate the virus. Thus, to reduce the risk of food-borne HEV transmission, pig livers must be thoroughly cooked.