scholarly journals Quantitative Proteomic Analyses of Influenza Virus-Infected Cultured Human Lung Cells

2010 ◽  
Vol 84 (20) ◽  
pp. 10888-10906 ◽  
Author(s):  
Kevin M. Coombs ◽  
Alicia Berard ◽  
Wanhong Xu ◽  
Oleg Krokhin ◽  
Xiaobo Meng ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Protein Function ◽  
Human Lung ◽  
Isotope Labeling ◽  
Morphological Changes ◽  
A549 Cells ◽  
Host Cells ◽  
Human Influenza Virus ◽  
Human Lung Cells ◽  
Cell Immunity

ABSTRACT Because they are obligate intracellular parasites, all viruses are exclusively and intimately dependent upon host cells for replication. Viruses, in turn, induce profound changes within cells, including apoptosis, morphological changes, and activation of signaling pathways. Many of these alterations have been analyzed by gene arrays, which measure the cellular “transcriptome.” Until recently, it has not been possible to extend comparable types of studies to globally examine all the host cellular proteins, which are the actual effector molecules. We have used stable isotope labeling by amino acids in cell culture (SILAC), combined with high-throughput two-dimensional (2-D) high-performance liquid chromatography (HPLC)/mass spectrometry, to determine quantitative differences in host proteins after infection of human lung A549 cells with human influenza virus A/PR/8/34 (H1N1) for 24 h. Of the 4,689 identified and measured cytosolic protein pairs, 127 were significantly upregulated at >95% confidence, 153 were significantly downregulated at >95% confidence, and a total of 87 proteins were upregulated or downregulated more than 5-fold at >99% confidence. Gene ontology and pathway analyses indicated differentially regulated proteins and included those involved in host cell immunity and antigen presentation, cell adhesion, metabolism, protein function, signal transduction, and transcription pathways.

scholarly journals The Hag Protein of Moraxella catarrhalis Strain O35E Is Associated with Adherence to Human Lung and Middle Ear Cells

2003 ◽  
Vol 71 (9) ◽  
pp. 4977-4984 ◽  
Author(s):  
Melissa M. Holm ◽  
Serena L. Vanlerberg ◽  
Darren D. Sledjeski ◽  
Eric R. Lafontaine
Keyword(s):  
Cell Lines ◽  
Middle Ear ◽  
Moraxella Catarrhalis ◽  
Human Lung ◽  
Primary Cultures ◽  
A549 Cells ◽  
Pcr Analysis ◽  
Nucleotide Sequence Data ◽  
Human Lung Cells ◽  
Conjunctival Cells

ABSTRACT Previous studies have demonstrated that the Moraxella catarrhalis surface antigen UspA1 is an adhesin for Chang human conjunctival cells. The present report demonstrates that lack of UspA1 expression does not affect the adherence of strain O35E to A549 human lung cells or primary cultures of human middle ear epithelial (HMEE) cells. These results imply that another molecule mediates the adherence of M. catarrhalis to these two cell lines. To identify this adhesin, strain O35E was mutagenized with a transposon and 1,000 mutants were screened in a microcolony formation assay using A549 cells. Nine independent isolates exhibited an 8- to 19-fold reduction in adherence and contained a transposon in the same locus. Nucleotide sequence data and PCR analysis indicated that the transposons were inserted in different locations in the gene encoding the surface protein Hag. Quantitative assays using one representative transposon mutant, O35E.TN2, showed considerably decreased binding to A549 as well as HMEE cells. However, this mutant adhered at wild-type levels to Chang conjunctival cells. These findings suggest that the M. catarrhalis Hag protein is an adhesin for cell lines derived from human lung and middle ear tissues.

scholarly journals Clarithromycin Inhibits Progeny Virus Production from Human Influenza Virus-Infected Host Cells

2008 ◽  
Vol 31 (2) ◽  
pp. 217-222 ◽  
Author(s):  
Daisei Miyamoto ◽  
Sayaka Hasegawa ◽  
Nongluk Sriwilaijaroen ◽  
Sangchai Yingsakmongkon ◽  
Hiroaki Hiramatsu ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Virus Production ◽  
Host Cells ◽  
Infected Host ◽  
Progeny Virus ◽  
Human Influenza ◽  
Human Influenza Virus ◽  
Progeny Virus Production

scholarly journals Metabolomic Analysis of Influenza A Virus A/WSN/1933 (H1N1) Infected A549 Cells during First Cycle of Viral Replication

Viruses ◽  
2019 ◽  
Vol 11 (11) ◽  
pp. 1007 ◽  
Author(s):  
Xiaodong Tian ◽  
Kun Zhang ◽  
Jie Min ◽  
Can Chen ◽  
Ying Cao ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Virus Infection ◽  
Influenza A Virus ◽  
Influenza A ◽  
Cell Metabolism ◽  
A549 Cells ◽  
Influenza Virus Infection ◽  
Tca Cycle ◽  
Host Cells ◽  
Pathway Analyses

Influenza A virus (IAV) has developed strategies to utilize host metabolites which, after identification and isolation, can be used to discover the value of immunometabolism. During this study, to mimic the metabolic processes of influenza virus infection in human cells, we infect A549 cells with H1N1 (WSN) influenza virus and explore the metabolites with altered levels during the first cycle of influenza virus infection using ultra-high-pressure liquid chromatography–quadrupole time-of-flight mass spectrometer (UHPLC–Q-TOF MS) technology. We annotate the metabolites using MetaboAnalyst and the Kyoto Encyclopedia of Genes and Genomes pathway analyses, which reveal that IAV regulates the abundance of the metabolic products of host cells during early infection to provide the energy and metabolites required to efficiently complete its own life cycle. These metabolites are correlated with the tricarboxylic acid (TCA) cycle and mainly are involved in purine, lipid, and glutathione metabolisms. Concurrently, the metabolites interact with signal receptors in A549 cells to participate in cellular energy metabolism signaling pathways. Metabonomic analyses have revealed that, in the first cycle, the virus not only hijacks cell metabolism for its own replication, but also affects innate immunity, indicating a need for further study of the complex relationship between IAV and host cells.

scholarly journals Aptamer Profiling of A549 Cells Infected with Low-Pathogenicity and High-Pathogenicity Influenza Viruses

Viruses ◽  
2019 ◽  
Vol 11 (11) ◽  
pp. 1028 ◽  
Author(s):  
Kevin M. Coombs ◽  
Philippe F. Simon ◽  
Nigel J. McLeish ◽  
Ali Zahedi-Amiri ◽  
Darwyn Kobasa
Keyword(s):  
Influenza A ◽  
Human Lung ◽  
A549 Cells ◽  
Influenza Viruses ◽  
Post Translational Modification ◽  
Emerging Pathogens ◽  
Influenza A Viruses ◽  
Human Lung Cells ◽  
Wide Range ◽  
High Pathogenicity

Influenza A viruses (IAVs) are important animal and human emerging and re-emerging pathogens that are responsible for yearly seasonal epidemics and sporadic pandemics. IAVs cause a wide range of clinical illnesses, from relatively mild infections by seasonal strains, to acute respiratory distress during infections with highly pathogenic avian IAVs (HPAI). For this study, we infected A549 human lung cells with lab prototype A/PR/8/34 (H1N1) (PR8), a seasonal H1N1 (RV733), the 2009 pandemic H1N1 (pdm09), or with two avian strains, an H5N1 HPAI strain or an H7N9 strain that has low pathogenicity in birds but high pathogenicity in humans. We used a newly-developed aptamer-based multiplexed technique (SOMAscan®) to examine >1300 human lung cell proteins affected by the different IAV strains, and identified more than 500 significantly dysregulated cellular proteins. Our analyses indicated that the avian strains induced more profound changes in the A549 global proteome compared to all tested low-pathogenicity H1N1 strains. The PR8 strain induced a general activation, primarily by upregulating many immune molecules, the seasonal RV733 and pdm09 strains had minimal effect upon assayed molecules, and the avian strains induced significant downregulation, primarily in antimicrobial response, cardiovascular and post-translational modification systems.

scholarly journals Cerium Oxide Nanoparticles Induced Toxicity in Human Lung Cells: Role of ROS Mediated DNA Damage and Apoptosis

2014 ◽  
Vol 2014 ◽  
pp. 1-14 ◽  
Author(s):  
Sandeep Mittal ◽  
Alok K. Pandey
Keyword(s):  
Dna Damage ◽  
Cell Death ◽  
Cerium Oxide ◽  
Oxidative Dna Damage ◽  
Apoptotic Cell ◽  
Morphological Changes ◽  
A549 Cells ◽  
Cerium Oxide Nanoparticles ◽  
Oxide Nanoparticles ◽  
Human Lung Cells

Cerium oxide nanoparticles (CeO2NPs) have promising industrial and biomedical applications. In spite of their applications, the toxicity of these NPs in biological/physiological environment is a major concern. Present study aimed to understand the molecular mechanism underlying the toxicity of CeO2NPs on lung adenocarcinoma (A549) cells. After internalization, CeO2NPs caused significant cytotoxicity and morphological changes in A549 cells. Further, the cell death was found to be apoptotic as shown by loss in mitochondrial membrane potential and increase in annexin-V positive cells and confirmed by immunoblot analysis of BAX, BCl-2, Cyt C, AIF, caspase-3, and caspase-9. A significant increase in oxidative DNA damage was found which was confirmed by phosphorylation of p53 gene and presence of cleaved poly ADP ribose polymerase (PARP). This damage could be attributed to increased production of reactive oxygen species (ROS) with concomitant decrease in antioxidant “glutathione (GSH)” level. DNA damage and cell death were attenuated by the application of ROS and apoptosis inhibitors N-acetyl-L- cysteine (NAC) and Z-DEVD-fmk, respectively. Our study concludes that ROS mediated DNA damage and cell cycle arrest play a major role in CeO2NPs induced apoptotic cell death in A549 cells. Apart from beneficial applications, these NPs also impart potential harmful effects which should be properly evaluated prior to their use.

Antagonistic effect of co-exposure to short-multiwalled carbon nanotubes and benzo[a]pyrene in human lung cells (A549)

2019 ◽  
Vol 35 (6) ◽  
pp. 445-456
Author(s):  
Mansour Rezazadeh Azari ◽  
Yousef Mohammadian ◽  
Habibollah Peirovi ◽  
Meisam Omidi ◽  
Fariba Khodagholi ◽  
...  
Keyword(s):  
Carbon Nanotubes ◽  
Multiwalled Carbon Nanotubes ◽  
Human Lung ◽  
A549 Cells ◽  
In Vitro Toxicity ◽  
Ros Generation ◽  
Exposure Condition ◽  
Multiwalled Carbon ◽  
Human Lung Cells ◽  
Starting Point

In theenvironment, co-exposure to short-multiwalled carbon nanotubes (S-MWCNTs) and polycyclic aromatic compounds (PAHs) has been reported. In the co-exposure condition, the adsorption of PAHs onto MWCNTs may reduce PAHs toxic effect. The objective of this study was to investigate the cytotoxicity of S-MWCNTs and benzo[a]pyrene (B[a]P) individually, and in combination in human lung cell lines (A549). The adsorption of B[a]P onto MWCNTs was measured spectrometrically. In vitro toxicity was assessed through cell viability, reactive oxygen species (ROS) generation, apoptosis, and 8-hydroxy-2′-deoxyguanosine (8-OHdG) generation experiments. The S-MWCNTs demonstrated cytotoxicity through the generation of ROS, apoptosis, and 8-OHdG in A549 cells. Co-exposure to S-MWCNTs and B[a]P demonstrated a significant reduction in ROS generation and apoptosis compared with the sum of their separate toxic effects at the same concentrations. Decreasing the bioavailability of B[a]P by MWCNT interaction is the probable reason for the antagonistic effects of the co-exposure condition. The findings of this study will contribute to a better understanding of the health effects of co-exposures to air pollutants and could be a starting point for modifying future health risk assessments.

scholarly journals In Vitro Cytotoxicity Evaluation of the Magnéli Phase Titanium Suboxides (TixO2x−1) on A549 Human Lung Cells

2019 ◽  
Vol 20 (1) ◽  
pp. 196 ◽  
Author(s):  
Veno Kononenko ◽  
Damjana Drobne
Keyword(s):  
Photocatalytic Activity ◽  
Tio2 Nanoparticles ◽  
Human Lung ◽  
A549 Cells ◽  
Cellular Interactions ◽  
Biological Interactions ◽  
Lysosomal Membranes ◽  
Human Lung Cells ◽  
Magnéli Phase ◽  
Magneli Phase

The use of titanium suboxides, known as Magnéli phase TiOx, is expected to increase in the near future due to their desirable properties. In order to use Magnéli phase TiOx nanoparticles safely, it is necessary to know how nanoparticles interact with biological systems. In this study, the cytotoxicity of three different Magnéli TiOx nanoparticles was evaluated using human lung A549 cells and the results were compared with hazard data on two different TiO2 nanoparticles whose biological interactions have already been extensively studied. After A549 cells were exposed to nanoparticles, the metabolic activity was measured by the Resazurin assay, the amount of cellular proteins was measured by the Coomassie Blue assay, and lysosomal integrity was measured by the Neutral Red Uptake assay. In order to investigate possible modes of particle actions, intracellular Ca2+ level, reactive oxygen species (ROS) production, and photo-oxidative disruptions of lysosomal membranes were assessed. All experiments were performed in serum-containing and in serum-deprived cell culture mediums. In addition, the photocatalytic activity of Magnéli TiOx and TiO2 nanoparticles was measured. The results show that Magnéli TiOx nanoparticles increase intracellular Ca2+ but not ROS levels. In contrast, TiO2 nanoparticles increase ROS levels, resulting in a higher cytotoxicity. Although Magnéli TiOx nanoparticles showed a lower UV-A photocatalytic activity, the photo-stability of the lysosomal membranes was decreased by a greater extent, possibly due to particle accumulation inside lysosomes. We provide evidence that Magnéli TiOx nanoparticles have lower overall biological activity when compared with the two TiO2 formulations. However, some unique cellular interactions were detected and should be further studied in line with possible Magnéli TiOx application. We conclude that Magnéli phase nanoparticles could be considered as low toxic material same as other forms of titanium oxide particles.

scholarly journals Global Human-Kinase Screening Identifies Therapeutic Host Targets against Influenza

2014 ◽  
Vol 19 (6) ◽  
pp. 936-946 ◽  
Author(s):  
Colm Atkins ◽  
Carrie W. Evans ◽  
Brian Nordin ◽  
Matthew P. Patricelli ◽  
Robert Reynolds ◽  
...  
Keyword(s):  
Influenza Virus ◽  
A549 Cells ◽  
Adenosine Diphosphate ◽  
Human Cells ◽  
Time Dependent ◽  
Small Interfering Rnas ◽  
Human Influenza Virus ◽  
Reverse Transcription Pcr ◽  
Novel Host ◽  
Dependent Signal

During viral infection of human cells, host kinases mediate signaling activities that are used by all viruses for replication; therefore, targeting of host kinases is of broad therapeutic interest. Here, host kinases were globally screened during human influenza virus (H1N1) infection to determine the time-dependent effects of virus infection and replication on kinase function. Desthiobiotin-labeled analogs of adenosine triphosphate and adenosine diphosphate were used to probe and covalently label host kinases in infected cell lysates, and probe affinity was determined. Using infected human A549 cells, we screened for time-dependent signal changes and identified host kinases whose probe affinities differed significantly when compared to uninfected cells. Our screen identified 10 novel host kinases that have not been previously shown to be involved with influenza virus replication, and we validated the functional importance of these novel kinases during infection using targeted small interfering RNAs (siRNAs). The effects of kinase-targeted siRNA knockdowns on replicating virus levels were measured by quantitative reverse-transcription PCR and cytoprotection assays. We identified several novel host kinases that, when knocked down, enhanced or reduced the viral load in cell culture. This preliminary work represents the first screen of the changing host kinome in influenza virus–infected human cells.

scholarly journals Antiandrogens Target TMPRSS2 and Reduce SARS-CoV-2 Virus Entry in Lung Cells

2021 ◽  
Vol 5 (Supplement_1) ◽  
pp. A60-A61
Author(s):  
Damien A Leach ◽  
Mohr Andrea ◽  
Ralf Zwacka ◽  
Stathis Giottis ◽  
Laura Yates ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Cell Surface ◽  
Human Lung ◽  
Surface Proteins ◽  
Host Cells ◽  
Mouse Lung ◽  
Lung Cells ◽  
Sequencing Data ◽  
Cell Surface Proteins ◽  
Human Lung Cells

Abstract The SARS-CoV-2 coronavirus is the cause of the COVID-19 pandemic. Entry of the virus into host cells, most destructively lung cells, requires two host cell surface proteins, ACE2 and TMPRSS2, downregulation of which is thus a potential therapeutic approach for COVID-19. Both of these cell surface proteins are steroid regulated: TMPRSS2 is a well-characterised androgen-regulated target in prostate cancer. Analysis of sequencing data shows co-expression of the androgen receptor (AR) and TMPRSS2 in key human lung cell types that are targeted by SARS- CoV-2. We show that treatment with antiandrogens such as enzalutamide (a well-tolerated drug widely used in advanced prostate cancer) significantly reduces TMPRSS2 levels in human lung cells and in vivo in mouse lung. We demonstrate that AR binding in the region of the TMPRSS2 gene differs between lung and prostate, identifying distinct regulatory regions. Together, the data and evidence presented supports clinical trials to assess the efficacy of antiandrogens as a treatment option for COVID-19.

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