Host immune and apoptotic responses to avian influenza virus H9N2 in human tracheobronchial epithelial cells.
The avian influenza virus H9N2 subtype has circulated in wild birds, is prevalent in domestic poultry, and has successfully crossed the species boundary to infect humans. Phylogenetic analyses showed that viruses of this subtype appear to have contributed to the generation of highly pathogenic H5N1 viruses. Little is known about the host responses to H9N2 viruses in human airway respiratory epithelium, the primary portal for viral infection. Using an apically differentiated primary human tracheobronchial epithelial (TBE) culture, we examined host immune responses to infection by an avian H9N2 virus, in comparison with a human H9N2 isolate. We found that IFN-β was the prominent antiviral component, whereas interferon gamma-induced protein 10 kDa (IP-10), chemokine (C-C motif) ligand (CCL)-5 and TNF-α may be critical in proinflammatory responses to H9N2 viruses. In contrast, proinflammatory IL-1β, IL-8, and even IL-6 may only play a minor role in pathogenicity. Apparently Toll-like receptor (TLR)-3, TLR-7, and melanoma differentiation-associated gene 5 (MDA-5) contributed to the innate immunity against the H9N2 viruses, and MDA-5 was important in the induction of IFN-β. We showed that the avian H9N2 virus induced apoptosis through the mitochondria/cytochrome c-mediated intrinsic pathway, in addition to the caspase 8-mediated extrinsic pathway, as evidenced by the cytosolic presence of active caspase 9 and cytochrome c, independent of truncated BH3 interacting domain death agonist (Bid) activation. Further, we demonstrated that FLICE-like inhibitory protein (FLIP), an apoptotic dual regulator, and the p53-dependent Bcl-2 family members, Bax and Bcl-x(s), appeared to be involved in the regulation of extrinsic and intrinsic apoptotic pathways, respectively. The findings in this study will further our understanding of host defense mechanisms and the pathogenesis of H9N2 influenza viruses in human respiratory epithelium.
Xing
Zheng
Z
Department of Population Health and Reproduction, School of Veterinary Medicine, University of California at Davis, Davis, CA, USA. zxing@umn.edu
Harper
Richart
R
Anunciacion
Jerome
J
Yang
Zengqi
Z
Gao
Wei
W
Qu
Bingqian
B
Guan
Yi
Y
Cardona
Carol J
CJ
D000818:Animals; D017209:Apoptosis / Q000235:genetics; D051023:BH3 Interacting Domain Death Agonist Protein / Q000378:metabolism; D001717:Birds; D001980:Bronchi / Q000276:immunology / Q000473:pathology / Q000821:virology*; D053446:CASP8 and FADD-Like Apoptosis Regulating Protein / Q000378:metabolism; D020169:Caspases / Q000378:metabolism; D002478:Cells, Cultured; D016207:Cytokines / Q000235:genetics / Q000378:metabolism; D003588:Cytopathogenic Effect, Viral; D053487:DEAD-box RNA Helicases / Q000235:genetics / Q000378:metabolism; D004789:Enzyme Activation; D004847:Epithelial Cells / Q000276:immunology / Q000473:pathology / Q000821:virology*; D005786:Gene Expression Regulation; D006801:Humans; D007113:Immunity, Innate / Q000235:genetics; D018836:Inflammation Mediators / Q000378:metabolism; D053127:Influenza A Virus, H9N2 Subtype / Q000254:growth & development / Q000472:pathogenicity*; D005585:Influenza in Birds / Q000276:immunology / Q000473:pathology / Q000821:virology*; D007251:Influenza, Human / Q000276:immunology / Q000473:pathology / Q000821:virology*; D000072640:Interferon-Induced Helicase, IFIH1; D034622:RNA Interference; D013997:Time Factors; D051193:Toll-Like Receptors / Q000235:genetics / Q000378:metabolism; D014132:Trachea / Q000276:immunology / Q000473:pathology / Q000821:virology*; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D051028:bcl-2-Associated X Protein / Q000378:metabolism; D051020:bcl-X Protein / Q000378:metabolism
C494826:BAX protein, human; C494811:BCL2L1 protein, human; D051023:BH3 Interacting Domain Death Agonist Protein; C494814:BID protein, human; D053446:CASP8 and FADD-Like Apoptosis Regulating Protein; D016207:Cytokines; D018836:Inflammation Mediators; C495901:TP53 protein, human; D051193:Toll-Like Receptors; D016159:Tumor Suppressor Protein p53; D051028:bcl-2-Associated X Protein; D051020:bcl-X Protein; D020169:Caspases; C452815:IFIH1 protein, human; D053487:DEAD-box RNA Helicases; D000072640:Interferon-Induced Helicase, IFIH1