Enterovirus 71 directly infects human natural killer cells and induces cell apoptosis

Enterovirus 71 (EV71) belongs to the Enterovirus genus of the family Picornaviridae and is the major causative agent of hand, foot and mouth disease (HFMD). Although clinical manifestations of HFMD are usually mild and self-limiting, severe HFMD patients suffer from a diverse array of neurological diseases and sometimes these diseases are fatal. HFMD usually occurs in young children and gradually becomes a new threaten in Asia. Unfortunately, effective EV71 vaccine is not available to date and alternative treatments are still in debate. This is partially due to the lack of understanding of EV71 pathogenesis and host immune responses against EV71. Natural killer (NK) cells are key effector cells in host antiviral activities by directly killing viral-infected cells and producing cytokines and chemokines, especially in early phase of viral infection. After enteroviruses infection, NK cells were one of the most abundant cell types in the inflammatory infiltrate, and appeared to limit both enteroviruses replication and virus-induced disease in experimental mice model. However, role of human NK cells during EV71 infection, especially the direct interaction between EV71 and human NK cells, was not studied extensively. Clinical observation manifested that patients with severe EV71 infection have marked diminished NK cells in peripheral blood. Therefore we hypothesized that EV71 might directly target human NK cells as one of its immunoevasion strategies. Here, we demonstrated for the first time that fresh primary human NK cells were susceptible to EV71 infection. By flow cytometry and florescence microscope, EV71 capsid protein VP1 was able to be detected in viral-infected NK cells as soon as 6 hours after infection and peaked at 24 hour after infection. In the same time, EV71 viral RNA was detected by quantitative RT-PCR and the viral copies increased from 6 hour onwards to peak at 12 hours after infection. We further demonstrated the infectious entry of EV71 in human NK cells was depended on clathrin-mediated endocytosis. Next, we illustrated that EV71 infection could trigger NK cells apoptosis as evidenced by increased Annexin V+, PI+, and activated caspase 3+ cells in EV71-treated NK cells. We further proved that the cytotoxicity of NK cells was inhibited by EV71 infection and this inhibition might not be related with down-regulation of NKp46, but may be related to the increased apoptosis. In conclusion, our data suggested that EV71 might directly target and kill NK cells as a strategy to evade human innate immunity, which might facilitate virus replication, transmission and then contribute to viral-related pathogenesis.published_or_final_versionPaediatrics and Adolescent MedicineMasterMaster of Philosoph

Data from: Expansion of genotypic diversity and establishment of 2009 H1N1 pandemic-origin internal genes in pigs in China

‘Two-way' transmission of influenza viruses between humans and swine has been frequently observed and the occurrence of the 2009 H1N1 pandemic influenza (pdm/09) demonstrated that swine-origin viruses could facilitate the genesis of a pandemic strain. Although multiple introductions to and reassortment in swine of the pdm/09 virus have been repeatedly reported in both Eurasia and the Americas, its long-term impact on the development of swine influenza viruses (SIVs) has not been systematically explored. Our comprehensive evolutionary studies on the complete genomes of 387 SIVs obtained from 2009 to 2012 in influenza surveillance in China revealed 17 reassortant genotypes with pdm/09-origin genes. Even though the entire 2009 pandemic virus and its surface genes cannot persist, its internal genes have becoming established and are now the predominant lineages in pigs in the region. The main persistent pdm/09-origin reassortant forms had at least 5 pdm/09-origin internal genes and their surface genes primarily of European avian-like (EA) or human H3N2-like SIV origin. These findings represent a marked change to the evolutionary patterns and ecosystem of SIVs in China. It is possible that the pdm/09-origin internal genes may be in the process of replacing EA- or triple reassortant-like internal genes. These alterations to the SIV gene pool need to be continually monitored to assess changes in the potential for SIVs to transmit to humans. Importance: Shortly after the emergence of the 2009 pandemic H1N1 (pdm/09) influenza virus, it was transmitted from humans to pigs and this continues to occur around the world. Many reassortants between pdm/09-origin viruses and enzootic swine influenza viruses (SIVs) have been detected. However, the long-term impact of pdm/09-origin viruses on the SIV gene pool, which could lead to the generation of influenza viruses with the potential to infect humans, has not been systematically examined. From extensive surveillance of SIVs over a 38-month period in southern China, it was found that, although neither complete pdm/09 viruses nor their surface genes could persist in pigs, their internal genes did persist. Over the survey period, these internal genes became predominant, potentially replacing those of the enzootic SIV lineages. The altered diversity of the SIV gene pool needs to be closely monitored for changes in the potential of SIVs to transmit to humans

The genesis and source of the H7N9 influenza viruses causing human infections in China

National Institutes of Health (National Institute of Allergy and Infectious Diseases) [HSN266200700005C]; Li Ka Shing Foundation; University Grants Committee of the Hong Kong SAR [AoE/M-12/06]; Shenzhen Peacock Plan High-End Talents Program [KQTD201203]; University Development Fund (HKU); Innovation and Technology Commission of the Hong Kong Government; Newton International Fellowship of the Royal Society; US Agency for International Development (USAID) Emerging Pandemic Threats Program, PREDICT project [GHN-A-OO-09-00010-00]; European Union [278433-PREDEMICS]; ERC [260864]; Wellcome Trust [092807]A novel H7N9 influenza A virus first detected in March 2013 has since caused more than 130 human infections in China, resulting in 40 deaths(1,2). Preliminary analyses suggest that the virus is a reassortant of H7, N9 and H9N2 avian influenza viruses, and carries some amino acids associated with mammalian receptor binding, raising concerns of a new pandemic(1,3,4). However, neither the source populations of the H7N9 outbreak lineage nor the conditions for its genesis are fully known(5). Using a combination of active surveillance, screening of virus archives, and evolutionary analyses, here we show that H7 viruses probably transferred from domestic duck to chicken populations in China on at least two independent occasions. We show that the H7 viruses subsequently reassorted with enzootic H9N2 viruses to generate the H7N9 outbreak lineage, and a related previously unrecognized H7N7 lineage. The H7N9 outbreak lineage has spread over a large geographic region and is prevalent in chickens at live poultry markets, which are thought to be the immediate source of human infections. Whether the H7N9 outbreak lineage has, or will, become enzootic in China and neighbouring regions requires further investigation. The discovery here of a related H7N7 influenza virus in chickens that has the ability to infect mammals experimentally, suggests that H7 viruses may pose threats beyond the current outbreak. The continuing prevalence of H7 viruses in poultry could lead to the generation of highly pathogenic variants and further sporadic human infections, with a continued risk of the virus acquiring human-to-human transmissibility

Maximum likelihood trees and genotype table of swine influenza virus

Maximum likelihood trees and genotype table of swine influenza viru

The genesis and source of the H7N9 influenza viruses causing human infections in China.

A novel H7N9 influenza A virus first detected in March 2013 has since caused more than 130 human infections in China, resulting in 40 deaths. Preliminary analyses suggest that the virus is a reassortant of H7, N9 and H9N2 avian influenza viruses, and carries some amino acids associated with mammalian receptor binding, raising concerns of a new pandemic. However, neither the source populations of the H7N9 outbreak lineage nor the conditions for its genesis are fully known. Using a combination of active surveillance, screening of virus archives, and evolutionary analyses, here we show that H7 viruses probably transferred from domestic duck to chicken populations in China on at least two independent occasions. We show that the H7 viruses subsequently reassorted with enzootic H9N2 viruses to generate the H7N9 outbreak lineage, and a related previously unrecognized H7N7 lineage. The H7N9 outbreak lineage has spread over a large geographic region and is prevalent in chickens at live poultry markets, which are thought to be the immediate source of human infections. Whether the H7N9 outbreak lineage has, or will, become enzootic in China and neighbouring regions requires further investigation. The discovery here of a related H7N7 influenza virus in chickens that has the ability to infect mammals experimentally, suggests that H7 viruses may pose threats beyond the current outbreak. The continuing prevalence of H7 viruses in poultry could lead to the generation of highly pathogenic variants and further sporadic human infections, with a continued risk of the virus acquiring human-to-human transmissibility