Proteome Alterations in Primary Human Alveolar Macrophages in Response to Influenza A Virus Infection

Abstract

In order to obtain a global picture of how alveolar macrophages respond to influenza A virus (IAV) infection, we used a quantitative proteomics method to systematically examine protein expression in the IAV-infected primary human alveolar macrophages. Of the 1214 proteins identified, 43 were significantly up-regulated and 63 significantly down-regulated at > 95% confidence. The expression of an array of interferon (IFN)-induced proteins was significantly increased in the IAV-infected macrophages. The protein with the greatest expression increase was ISG15, an IFN-induced protein that has been shown to play an important role in antiviral defense. Concomitantly, quantitative real time PCR analysis revealed that the gene expression of type I IFNs increased substantially following virus infection. Our results are consistent with the notion that type I IFNs play a vital role in the response of human alveolar macrophages to IAV infection. In addition to the IFN-mediated responses, inflammatory response, apoptosis and redox state rebalancing appeared also to be major pathways that were affected by IAV infection. Furthermore, our data suggest that alveolar macrophages may play a crucial role in regenerating alveolar epithelium during IAV infection.

Proteomic analysis reveals down-regulation of surfactant protein B in murine type II pneumocytes infected with influenza A virus

“…In a previous study, PR8 replication in primary murine ATII cells peaked at 24 h (Kebaabetswe et al, 2013). This time point is consistent with other proteomic studies using the same strain of influenza A virus in different cell types, including primary cultures (Coombs et al, 2010; Kroeker et al, 2012; Liu et al, 2012). …”

“…Proteins that form the viral RNA-dependent RNA polymerase complex, PB1, PB2, and PA, the nuclear export protein, NEP, and M2 protein are expressed in relatively low abundance in infected cells (Kummer et al, 2014) and were not detected by our analysis. Overall, we detected a similar profile of viral proteins as others have reported in proteomics analyses of influenza virus- infected cells and animals (Brown et al, 2010; Lietzen et al, 2011; Liu et al, 2012). …”

“…Proteomics studies of influenza virus infections in macaques (Brown et al, 2010), mice (Kumar et al, 2014), continuous cell lines (Coombs et al, 2010; Kummer et al, 2014; Vester et al, 2009), and primary human cells (Kroeker et al, 2012; Lietzen et al, 2011; Liu et al, 2012) have provided insight into how IAV strains alter the host proteome at cellular and organismal levels. However, host responses to IAV infection are also, in part, specific to the cell type.…”

Influenza A Virus-induced expression of ISG20 inhibits viral replication by interacting with nucleoprotein

“…To date, high-throughput RNA sequencing (RNA-seq) has been the most valuable method developed to systemically explore transcriptomics. Thousands of genes that undergo transcriptional regulation after IAV infection have been identified via RNA-seq by several groups [9][10][11][12]. Gene ontology enrichment and pathway analysis demonstrate that the regulated genes are involved in various biological processes, including immune responses, cell adhesion, cell metabolism, antigen presentation, and signal transduction.…”

Quantitative intact proteomics investigations of alveolar macrophages in sarcoidosis

“…While these investigations provide an important basis for understanding of macrophage differentiation and polarisation occurring in the lung, global proteomics studies investigating phenotypic alterations of human primary alveolar macrophages in response to respiratory disease are scarce. Exceptions include investigations of alveolar macrophage proteome responses to infection by porcine reproductive and respiratory syndrome virus [13] and influenza A virus [14]. Previous proteomics studies on sarcoidosis have been performed mainly on bronchoalveolar lavage (BAL) fluid and serum [15][16][17][18][19][20][21][22], thereby primarily reflecting proteins actively secreted or exudated into the bronchoalveolar lumen, such as plasma proteins and antioxidant proteins.…”