The miRNA expression patterns in WSN-infected A549 cells at various time points post-infection.
ABSTRACT: To identify miRNAs that are affected by IAV infection, we adopted a systematic approach, and performed a miRNA microarray analysis using a human alveolar adenocarcinoma cell line, A549, which was infected with a common laboratory strain of influenza A/WSN/33(H1N1) virus (WSN). We compared miRNA expression profiles in A549 cells infected with WSN for 2, 6, and 10 hours at a multiplicity of infection (MOI) of 2. Among 955 miRNAs on the microarray, 209 miRNAs were detected in all three infected A549 samples, but when compared with analysis results from mock-infected A549 cells, most expression changes were less than 1.5-fold , which was consistent with a previous report (Buggele et al., 2013). Therefore, we established 1.5-fold expression change as a threshold to select for miRNAs potentially affected by WSN infection. Further screening revealed 116 miRNAs that underwent 1.5-fold or greater expression changes at any timepoint following WSN infection. Overall design: The A549 cells were infected with WSN at an MOI of 2 in serum-free MEM medium for 2, 6 and 10 hours.
INSTRUMENT(S): Agilent-021827 Human miRNA Microarray (V3) (Probe Name version)
Project description:Whole-genome data was developed from influenza virus infected A549 cells to better characterize the effect of C646 on influenza virus infection A549 cells were treated with C646 or DMSO for 10 h, and then were infected with A/WSN/33 virus (WSN; H1N1) at a multiplicity of infection (MOI) 2. A549 cells for microarray studies were collected at different times. The gene expression in A549 cells was compared between C646-treated group and DMSO-treated group.
Project description:A growing body of evidence suggests gene regulatory functions for the majority of non-protein-coding RNAs (ncRNAs). Besides small RNAs (sRNAs), the diverse class of long ncRNAs (lncRNAs) recently came into focus of research. So far, the relevance of lncRNAs in infection processes remains elusive. Here, we report the differential expression of several classes of lncRNAs during influenza A virus (IAV) infection in human lung epithelial cells. 2 biological replicates of each condition were hybridzed in an independent color-swap; A549 cells were washed with PBS and then infected with viruses at MOI 1 in infection buffer for for 60 min at room temperature. Cells were incubated for the indicated time periods at 37 °C in DMEM supplemented with 0.2% bovine serum albumin, 4 mM l-glutamine and antibiotics. Supernatants of A/WSN/33 (H1N1) virus infected A549 cells (MOI 5, 4hpi) were exposed to UV light for 5 min and then used to stimulate A549 cells for 4 h Supernatants of A/WSN/33 (H1N1) virus infected A549 cells (MOI 5, 8hpi) were exposed to UV light for 5 min and then used to stimulate A549 cells for 8 h
Project description:To investigate whether influenza A virus (IAV) infection alters cellular miRNA profiles, A549 cells were either mock-infected or infected with two different subtypes of IAV, A/Beijing/501/2009 (H1N1; MOI=5) and A/Vietnam/1194/2004 (H5N1; MOI=2), for 24 or 48 h. Total RNA was purified from cell samples, and microarray analysis of miRNAs was performed. The expression of selected miRNAs was further confirmed by quantitative real-time PCR. Overall design: A549 cells were mock-infected or infected with two different subtypes of IAV, A/Beijing/501/2009 (H1N1; MOI=5) and A/Vietnam/1194/2004 (H5N1; MOI=2), for 24 or 48 h. Then total RNA was purified from cell samples to measure the microRNAs expression.
Project description:We used the microarray data to analyze host cells response on A549 cells infected with A/WSN/33 (H1N1) The A/WSN/33 (H1N1) infected A549 cells were harvested at 10 hpi and RNA extraction was performed using standard protocol as described by Affymetrix. The aim of this experiment is to analyze host response to Influenza A/WSN/33 (H1N1) infection.
Project description:We used the microarray data to analyze host cells response on A549 cells infected with A/WSN/33 (H1N1) The A/WSN/33 (H1N1) infected A549 cells were harvested at 2, 4 and 6 hpi and RNA extraction was performed using standard protocol as described by Affymetrix. The aim of this experiment is to analyze host response to Influenza A/WSN/33 (H1N1) infection.
Project description:Human lncRNA NRAV-overexpression in A549 cells was found to increase the influenza virus WSN replication. To discover the mechanism underlying this promotion by NRAV, genome-wide mRNA expression was measured by using microarray. Thousands of genes were differentially expressed and some ISGs were down-regulated. Human lncRNA NRAV-overexpressioning A549 cells (Test) and empty vector control cells were infected with influenza virus A/WSN/33 (H1N1) for 16 hours at a MOI of 3. Six total RNA samples from three independent experiments were extracted and used for mRNA microarray.
Project description:The goal of this experiment was to determine gene expression changes during influenza A virus infection as the result of expression influenza virus inducible miRNAs in A549 cells. The gene expression profiling experiment was performed with 4 groups (mock infected, influenza A virus infected, influenza A virus infected in the presence of exogenous miR-141, miR-374b, miR-449b, miR-518b, and miR-1263, and influenza A virus infected in the presence of exogenous miR-147b, miR-190b, miR-199a, miR-512-5p, and miR-874 with 3 biological replicates for each group. Total RNA was purified from A549 cells that were mock infected or infected with influenza A virus (A/WSN/33, 5pfu/cell) alone or in the presence of miRNA mimics 10 hours after treatment.
Project description:We identified the abundance of isoforms of miRNAs encoded by human adenovirus 5 in total cytoplasmic RNA fractions of infected cells as well as in fractions containing the purified RNA-induced silencing complex (RISC). Overall design: Human A549 cells were infected with human adenovirus 5 at an MOI of 30 TCID50/cell or were mock-infected. Total RNA was isolated at 30h post-infection. In addition, RNA incorporated in RISC which was isolated by immunoprecipitation using an antibody recognizing all human Argonaute proteins was conducted as well. Purified RNAs were used to generate sRNA libraries with were subjected to RNA sequencing.
Project description:To further understand the roles of miRNA during influenza A virus infection, we performed miRNA profiling in human alveolar adenocarcinoma cell lines, A549 cells, infected with influenza A virus A/Beijing/501/2009(H1N1) and A/goose/Jilin/hb/2003(H5N1). Overall design: A549 cells were either mock-infected or infected with influenza virus H1N1 or H5N1 at a MOI of 5. Total RNAs were isolated at 8 and 24 h post-infection and analysed for miRNA expression profiles. Each group was run in triplicate.
Project description:Analysis of gene expression in human macrophages infected with influenza A viruses expressing full length or truncated NS1 protein. The hypothesis tested was that C-terminal truncations of viral NS1 protein attenuate the capability of NS1 to limit activation of host antiviral and immune response genes. Cells were infected with influenza A/WSN/33 viruses expressing wild type NS1 protein (WSN-230), NS1 protein of 220 aa long (WSN-220) and NS1 protein of 202 aa long (WSN-202) on non-infected (Mock) Overall design: Total RNA isolated from macrophages after 8 hours of infection with wild type or mutant influenza A virus (multiplicity of infection = 2)