Inhibition of Autophagy Ameliorates Acute Lung Injury Caused by Avian Influenza A H5N1 Infection
Preventing H5N1 from Damaging Lungs
The higher mortality rate of the H5N1 strain of avian influenza compared to that of the seasonal H1N1 virus is attributed to the more severe lung damage caused by the H5N1 strain. Sun et al. found that lung tissue from an individual infected with H5N1 contained many autophagosomes and that mice infected with H5N1 had greater numbers of autophagosomes in lung tissue than did mice infected with the H1N1 strain. In addition, the H5N1 virus stimulated autophagic signaling in mouse epithelial cells to a greater extent than did the H1N1 virus. Pharmacological inhibition of autophagic signaling or knockdown of components of the autophagy pathway in H5N1-infected mice resulted in less severe lung damage, increased survival rate, and decreased mortality. These findings suggest that targeting the autophagy pathway might provide therapeutic targets in treating H5N1 infection in humans.
Abstract
The threat of a new influenza pandemic has existed since 1997, when the highly pathogenic H5N1 strain of avian influenza A virus infected humans in Hong Kong and spread across Asia, where it continued to infect poultry and people. The human mortality rate of H5N1 infection is about 60%, whereas that of seasonal H1N1 infection is less than 0.1%. The high mortality rate associated with H5N1 infection is predominantly a result of respiratory failure caused by acute lung injury; however, how viral infection contributes to this disease pathology is unclear. Here, we used electron microscopy to show the accumulation of autophagosomes in H5N1-infected lungs from a human cadaver and mice, as well as in infected A549 human epithelial lung cells. We also showed that H5N1, but not seasonal H1N1, induced autophagic cell death in alveolar epithelial cells through a pathway involving the kinase Akt, the tumor suppressor protein TSC2, and the mammalian target of rapamycin. Additionally, we suggest that the hemagglutinin protein of H5N1 may be responsible for stimulating autophagy. When applied prophylactically, reagents that blocked virus-induced autophagic signaling substantially increased the survival rate of mice and substantially ameliorated the acute lung injury and mortality caused by H5N1 infection. We conclude that the autophagic cell death of alveolar epithelial cells likely plays a crucial role in the high mortality rate of H5N1 infection, and we suggest that autophagy-blocking agents might be useful as prophylactics and therapeutics against infection of humans by the H5N1 virus.
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Supplementary Material
Summary
Fig. S1. Autophagy is induced by H5N1 infection of mouse lungs and A549 cells.
Fig. S2. LC3 puncta occur in H5N1-infected A549 cells and decrease in abundance upon treatment with TSC2-specific siRNA.
Fig. S3. Autophagic death of A549 cells induced by H5N1 infection is substantially rescued by Atg6-specific siRNA.
Fig. S4. The production of proinflammatory cytokines by H5N1 infection is not altered by Atg5-specific siRNA.
Fig. S5. The increased immune response induced by H5N1 infection is not altered by Atg5-specific siRNA.
Fig. S6. Mouse model of acute lung injury induced by infection with live H5N1 virus.
Fig. S7. Acute lung injury in mice induced by live H5N1 virus was ameliorated by Atg6-specific siRNA.
Fig. S8. Acute lung injury in mice induced by live H5N1 virus was not ameliorated by the selective inhibitors of apoptosis or necrosis.
Fig. S9. Mice infected by H1N1 virus and treated with Atg5-specific siRNA do not develop acute lung injury.
Fig. S10. Rag1 knockout mice develop acute lung injury induced by H5N1 virus.
Fig. S11. The HA protein of H5N1 virus may induce autophagy in human A549 cells.
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Published In
Science Signaling
Volume 5 | Issue 212
February 2012
February 2012
Copyright
Copyright © 2012, American Association for the Advancement of Science.
Submission history
Received: 17 February 2011
Accepted: 2 February 2012
Acknowledgments
We thank Z. Han, W. Xi, H. Wang, R. Lu, X. Chen, Z. Pan, R. Sheng, and X. Liu for technical support; H. Zhang and J. Ma for experiments performed in mouse embryonic fibroblast cells with formulated chicken inactivated virus vaccines; and J. Penninger, L. Li, and H. Pickersgill for helpful discussions and for editing the manuscript. Funding: This work was supported by the National Natural Science Foundation of China (30625013, 30623009, and 81000764), the Ministry of Science and Technology of China (2009CB522105), and the Ministry of Health (2009ZX10004-308). D.L. acknowledges support by the Science and Technology Commission of Shanghai Municipality (07pj14096). Author contributions: Y. Sun, C.L., X.J., Z.Z., H.W., S.R., F.G., W.N., Y.Z., Y.Y., J.T., and C.Z. performed the experiments; Y. Shu and R.G. provided H5N1-infected human lung tissue; H. Liu, P.Y., K.L., and S.W. constructed the H5-expressing cell line and expressed and purified the H5 protein; H. Lu, X.L., and L.T. prepared live virus; J.S. took EM photos of human tissue; X.G. provided Rag1 knockout mice; X.T., Y.Q., and K.-F.X. provided normal human lung tissue; D.L. provided helpful ideas; C.J. and N.J. designed the experiments and analyzed the data; and C.J. wrote the manuscript. Competing interests: C.J., S.R., H. Liu, F.G., H.W., Y. Sun, and C.L. have filed for a patent in China for the use of autophagy inhibitors in the prevention or treatment of acute lung injury. The file number: PCT/CN2010/072331.
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