title: Ion-Current-Based Temporal Proteomic Profiling of Influenza-A-Virus-Infected Mouse Lungs Revealed Underlying Mechanisms of Altered Integrity of the Lung Microvascular Barrier.
abstract: Investigation of influenza-A-virus (IAV)-infected lung proteomes will greatly promote our understanding on the virus-host crosstalk. Using a detergent-cocktail extraction and digestion procedure and a reproducible ion-current-based method, we performed the first comprehensive temporal analysis of mouse IAV infection. Mouse lung tissues at three time points post-inoculation were compared with controls (n = 4/group), and >1600 proteins were quantified without missing value in any animal. Significantly changed proteins were identified at 4 days (n = 144), 7 days (n = 695), and 10 days (n = 396) after infection, with low false altered protein rates (1.73-8.39%). Functional annotation revealed several key biological processes involved in the systemic host responses. Intriguingly, decreased levels of several cell junction proteins as well as increased levels of tissue metalloproteinase MMP9 were observed, reflecting the IAV-induced structural breakdown of lung epithelial barrier. Supporting evidence of MMP9 activation came from immunoassays examining the abundance and phosphorylation states of all MAPKs and several relevant molecules. Importantly, IAV-induced MMP gelatinase expression was suggested to be specific to MMP9, and p38 MAPK may contribute predominantly to MMP9 elevation. These findings help to resolve the long-lasting debate regarding the signaling pathways of IAV-induced MMP9 expression and shed light on the molecular mechanisms underlying pulmonary capillary-alveolar leak syndrome that can occur during influenza infection.
authors.affiliation: New York State Center of Excellence in Bioinformatics & Life Sciences , 701 Ellicott Street, Buffalo, New York 14203, United States.|Jacobs School of Medicine and Biomedical Sciences, SUNY at Buffalo , South Campus, Buffalo, New York 14214, United States.
authors.forename: Shichen
authors.initials: S
authors.lastname: Shen
authors.affiliation: Department of Pharmaceutical Sciences, SUNY at Buffalo , South Campus, Buffalo, New York 14214, United States.|New York State Center of Excellence in Bioinformatics & Life Sciences , 701 Ellicott Street, Buffalo, New York 14203, United States.
authors.forename: Jun
authors.initials: J
authors.lastname: Li
authors.affiliation: Division of Nephrology, University of Rochester Medical Center , 601 Elmwood Avenue, Rochester, New York 14642, United States.
authors.forename: Shannon
authors.initials: S
authors.lastname: Hilchey
authors.affiliation: New York State Center of Excellence in Bioinformatics & Life Sciences , 701 Ellicott Street, Buffalo, New York 14203, United States.|Jacobs School of Medicine and Biomedical Sciences, SUNY at Buffalo , South Campus, Buffalo, New York 14214, United States.
authors.forename: Xiaomeng
authors.initials: X
authors.lastname: Shen
authors.affiliation: Department of Pharmaceutical Sciences, SUNY at Buffalo , South Campus, Buffalo, New York 14214, United States.|New York State Center of Excellence in Bioinformatics & Life Sciences , 701 Ellicott Street, Buffalo, New York 14203, United States.
authors.forename: Chengjian
authors.initials: C
authors.lastname: Tu
authors.affiliation: Department of Biostatistics and Computational Biology, University of Rochester , 265 Crittenden Boulevard, Rochester, New York 14642, United States.
authors.forename: Xing
authors.initials: X
authors.lastname: Qiu
authors.affiliation: Jacobs School of Medicine and Biomedical Sciences, SUNY at Buffalo , South Campus, Buffalo, New York 14214, United States.
authors.forename: Andrew
authors.initials: A
authors.lastname: Ng
authors.affiliation: Department of Biology, University of Rochester , 402 Hutchison Hall, Rochester, New York 14627, United States.
authors.forename: Sina
authors.initials: S
authors.lastname: Ghaemmaghami
authors.affiliation: Department of Biostatistics, School of Public Health, University of Texas Health Science Center at Houston , 1200 Pressler Street, Houston, Texas 77030, United States.
authors.forename: Hulin
authors.initials: H
authors.lastname: Wu
authors.affiliation: Division of Nephrology, University of Rochester Medical Center , 601 Elmwood Avenue, Rochester, New York 14642, United States.
authors.forename: Martin S
authors.initials: MS
authors.lastname: Zand
authors.affiliation: Department of Pharmaceutical Sciences, SUNY at Buffalo , South Campus, Buffalo, New York 14214, United States.|New York State Center of Excellence in Bioinformatics & Life Sciences , 701 Ellicott Street, Buffalo, New York 14203, United States.
authors.forename: Jun
authors.initials: J
authors.lastname: Qu
mesh_terms: D000818:Animals; D015824:Blood-Air Barrier / Q000378:metabolism* / Q000821:virology; D015153:Blotting, Western; D056148:Chromatography, Reverse-Phase; D053122:Influenza A Virus, H3N2 Subtype / Q000502:physiology; D016014:Linear Models; D008168:Lung / Q000098:blood supply / Q000378:metabolism* / Q000821:virology; D008297:Male; D013058:Mass Spectrometry; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D008810:Mice, Inbred C57BL; D009976:Orthomyxoviridae Infections / Q000378:metabolism* / Q000821:virology; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*
chemical_list: D020543:Proteome; D020780:Matrix Metalloproteinase 9
keywords: bottom-up proteomics; host factors; influenza; ion-current-based quantification; microvascular barrier