Human Lymphocyte Apoptosis after Exposure to Influenza A Virus

128

PA-X is a newly discovered protein that decreases the virulence of the 1918 H1N1 virus in a mouse model. However, the role of PA-X in the pathogenesis of highly pathogenic avian influenza viruses (HPAIV) of the H5N1 subtype in avian species is totally unknown. By generating two PA-X-deficient viruses and evaluating their virulence in different animal models, we show here that PA-X diminishes the virulence of the HPAIV H5N1 strain A/Chicken/Jiangsu/k0402/2010 (CK10) in mice, chickens, and ducks. Expression of PA-X dampens polymerase activity and virus replication both in vitro and in vivo. Using microarray analysis, we found that PA-X blunts the global host response in chicken lungs, markedly downregulating genes associated with the inflammatory and cell death responses. Correspondingly, a decreased cytokine response was recapitulated in multiple organs of chickens and ducks infected with the wild-type virus relative to those infected with the PA-X-deficient virus. In addition, the PA-X protein exhibits antiapoptotic activity in chicken and duck embryo fibroblasts. Thus, our results demonstrated that PA-X acts as a negative virulence regulator and decreases virulence by inhibiting viral replication and the host innate immune response. Therefore, we here define the role of PA-X in the pathogenicity of H5N1 HPAIV, furthering our understanding of the intricate pathogenesis of influenza A virus. Influenza A virus (IAV) can infect diverse host species, from wild and domestic birds to mammalian species, and the pathogenesis of IAV is complex due to its remarkable genetic variability. The genome of IAV contains eight RNA segments that encode at least 17 viral proteins, including 8 initially identified proteins (PB2, PB1, PA, HA, NP, NA, M1, and NS1), 2 splicing variants of the M and NS genes (M2 and NS2) (1-3), and the recently identified proteins PB1-N40 (4), PB1-F2 (5), PA-X (6), M42 (7), NS3 (8), PA-N155, and PA-N182 (9). PB1-N40 is an N-terminally truncated version of the PB1 protein that lacks the transcriptase function but can still interact with other polymerase complex subunits and regulate virus replication in a specific genetic background (4). PB1-F2, encoded by an alternative open reading frame (ORF) of PB1, has multiple functions, including the induction of apoptosis (10), aggravation of inflammation (11,12), and secondary bacterial infection (13). PA-X is a frameshift product of the ribosome and acts to decrease the virulence of the 1918 H1N1 virus in mice (6). PA-N155 and PA-N182 are N-terminally truncated forms of PA, and the mutant viruses lacking these two proteins exhibit attenuated in vitro replication and pathogenicity in mice relative to the wild-type (wt) virus (9). M42 is the M2 isoform with an alternative ectodomain that can functionally replace M2 and support efficient viral replication (7). Selman et al. have identified NS3 as the isoform of NS1 and speculated that the codon providing NS3 expression could be associated with host adaptation and the overcoming of the species barrier (8)....

Section: Discussionmentioning

confidence: 99%

PA-X Decreases the Pathogenicity of Highly Pathogenic H5N1 Influenza A Virus in Avian Species by Inhibiting Virus Replication and Host Response

Wang

et al. 2015

128

“…Therefore, any incompatibility between the requirements of the virus and what is available in the host cell might lead to an abortive infection. In fact, as with NK cells, the lymphocytes in general could not support viral replication, although they express viral protein with the help of macrophages (24,27). In mammals, replication-competent influenza A viruses are generally recovered from the superficial epithelium of the respiratory tract (9), where the viruses replicate and spread.…”

Section: Discussionmentioning

confidence: 99%

Influenza Virus Directly Infects Human Natural Killer Cells and Induces Cell Apoptosis

Mao

Qin

et al. 2009

131

126

Influenza is an acute respiratory viral disease that is transmitted in the first few days of infection. Evasion of host innate immune defenses, including natural killer (NK) cells, is important for the virus's success as a pathogen of humans and other animals. NK cells encounter influenza viruses within the microenvironment of infected cells and are important for host innate immunity during influenza virus infection. It is therefore important to investigate the direct effects of influenza virus on NK cells. In this study, we demonstrated for the first time that influenza virus directly infects and replicates in primary human NK cells. Viral entry into NK cells was mediated by both clathrin-and caveolin-dependent endocytosis rather than through macropinocytosis and was dependent on the sialic acids on cell surfaces. In addition, influenza virus infection induced a marked apoptosis of NK cells. Our findings suggest that influenza virus can directly target and kill NK cells, a potential novel strategy of influenza virus to evade the NK cell innate immune defense that is likely to facilitate viral transmission and may also contribute to virus pathogenesis.

“…These data show that NDV kills cancer cells primarily by activation of the mitochondrial death pathway, and furthermore, they indicate that the caspase-8/Bid-dependent signal amplification loop is not important for NDV-induced death. Viruses that induce death receptor-dependent apoptosis include human immunodeficiency virus (38), measles virus (57), influenza A virus (40), Sindbis virus (16), reovirus (9), lyssavirus (23), and hepatitis C virus (2). A number of viruses have been found to cause relocalization of proapoptotic mitochondrial proteins into the cytosol.…”

Section: Discussionmentioning

confidence: 99%

Newcastle Disease Virus Exerts Oncolysis by both Intrinsic and Extrinsic Caspase-Dependent Pathways of Cell Death

Elankumaran

Rockemann

Samal

2006

194

197

Newcastle disease virus (NDV), an avian paramyxovirus, is tumor selective and intrinsically oncolytic. Here, we present evidence that genetically modified, recombinant NDV strains are cytotoxic to human tumor cell lines of ecto-, endo-, and mesodermal origin. We show that cytotoxicity against tumor cells is due to multiple caspase-dependent pathways of apoptosis independent of interferon signaling competence. The signaling pathways of NDV-induced, cancer cell-selective apoptosis are not well understood. We demonstrate that NDV triggers apoptosis by activating the mitochondrial/intrinsic pathway and that it acts independently of the death receptor/extrinsic pathway. Caspase-8-methylated SH-SY5Y neuroblastoma cells are as sensitive to NDV as other caspase-8-competent cells. This demonstrates that NDV is likely to act primarily through the mitochondrial death pathway. NDV infection results in the loss of mitochondrial membrane potential and the subsequent release of the mitochondrial protein cytochrome c, but the second mitochondrion-derived activator of caspase (Smac/DIABLO) is not released. In addition, we describe early activation of caspase-9 and caspase-3. In contrast, cleavage of caspase-8, which is predominantly activated by the death receptor pathway, is a TNF-related, apoptosis-inducing ligand (TRAIL)-induced late event in NDV-mediated apoptosis of tumor cells. Our data, therefore, indicate that the death signal(s) generated by NDV in tumor cells ultimately converges at the mitochondria and that it acts independently of the death receptor pathway. Our cytotoxicity studies demonstrate that recombinant NDV could be developed as a cancer virotherapy agent, either alone or in combination with therapeutic transgenes. We have also shown that trackable oncolytic NDV could be developed without any reduction in oncolytic efficacy.