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ITRAQ-based comparative proteomic analysis of cells infected with Eimeria tenella sporozoites.

ABSTRACT: Eimeria tenella is an obligate intracellular parasite that actively invades cecal epithelial cells of chickens. When E. tenella infects a host cell, the host produces a corresponding change to deal with damage caused by this infection. To date, our knowledge on the mechanism of how the host cell responds to E. tenella infection is highly limited at both the molecular and cellular levels. In this study, isobaric tags for relative and absolute quantitation (iTRAQ) coupled with LC-MS/MS was used to screen the differentially expressed proteins (DEPs) in BHK-21 cells infected with E. tenella sporozoites for 24 h post infection. In total, 6139 non-redundant distinct proteins were identified and 195 of these were found to have a fold change ratio ?1.3 or ?0.7 and p < 0.05, including 151 up-regulated proteins and 44 down-regulated proteins. The reliability of the proteomic data was further validated with qPCR and western blot. Gene Ontology enrichment indicated that the up-regulated DEPs were mainly involved in binding and catalytic activity, whereas the down-regulated DEPs were catalytic activity and molecular function regulators. Furthermore, KEGG pathway analysis showed that the DEPs participated in the PI3K-Akt, chemokine, Ras, Wnt, and p53 signaling pathways and so on, and the up-regulated and down-regulated DEPs mainly related to the ribosome and mRNA surveillance pathway, respectively. The data in this study provide an important basis to further analyze E. tenella host cell interactions.

SUBMITTER: Zhao Z

PROVIDER: S-EPMC6383524 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature

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iTRAQ-based comparative proteomic analysis of cells infected with Eimeria tenella sporozoites.

Zhao Zongping Z Zhao Qiping Q Zhu Shunhai S Huang Bing B Lv Ling L Chen Ting T Yan Ming M Han Hongyu H Dong Hui H

Parasite (Paris, France) 20190220

Eimeria tenella is an obligate intracellular parasite that actively invades cecal epithelial cells of chickens. When E. tenella infects a host cell, the host produces a corresponding change to deal with damage caused by this infection. To date, our knowledge on the mechanism of how the host cell responds to E. tenella infection is highly limited at both the molecular and cellular levels. In this study, isobaric tags for relative and absolute quantitation (iTRAQ) coupled with LC-MS/MS was used to ...[more]

PMID: 30789155

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Project description:The re-emerging porcine epidemic diarrhea virus (PEDV) variant related diarrhea has been documented in China since late 2010 and now with global distribution. Currently, a virulent PEDV CH/YNKM-8/2013 and a CV777 vaccine strain-like AH-M have been successfully isolated from the clinical samples. To dissect out the underlying pathogenic mechanism of virulent PEDV and clarify the differences between virulent and CV777 vaccine strain-like PEDV infections, we performed an iTRAQ-based comparative quantitative proteomic study of Vero cells infected with both PEDV strains. A total of 661 and 474 differentially expressed proteins were identified upon virulent and CV777 vaccine strain-like isolates infection, respectively. Ingenuity Pathway Analysis was employed to investigate the canonical pathways and functional networks involved in both PEDV infections. Comprehensive studies have revealed that the PEDV virulent strain suppressed protein synthesis of Vero cells through down-regulating mTOR as well as its downstream targets 4EBP1 and p70S6K activities, which were validated by immunoblotting. In addition, the virulent strain could activate NF-κB pathway more intensively than the CV777 vaccine strain-like isolate, and elicit stronger inflammatory cascades as well. These data might provide new insights for elucidating the specific pathogenesis of PEDV infection, and pave the way for the development of effective therapeutic strategies.Biological significancePorcine epidemic diarrhea is now worldwide distributed and causing huge economic losses to swine industry. The immunomodulation and pathogenesis between PEDV and host, as well as the difference between virulent and attenuated strains of PEDV infections are still largely unknown. In this study, we presented for the first application of proteomic analysis to compare whole cellular protein alterations induced by virulent and CV777 vaccine strain-like PEDV infections, which might contribute to understand the pathogenesis of PEDV and anti-viral strategy development.

Project description:BackgroundAvian coccidiosis is an important parasitic disease that has serious adverse effects on the global poultry industry. The extensive use of anticoccidial drugs has resulted in an increase in drug resistance. Ethanamizuril (EZL) is a novel triazine with high anticoccidial activity.MethodsWe compared oocyst production and sporulation between EZL-sensitive (S) and EZL-resistant Eimeria tenella strains (R10 and R200) and used label-free quantitative proteomics to identify differentially expressed proteins (DEPs) between these strains.ResultsWe generated two EZL-resistant E. tenella strains: strain R10, which was induced using a constant dose of 10 mg EZL/kg poultry feed, and strain R200, which was generated by gradually increasing the EZL dosage to 200 mg EZL/kg poultry feed. With an increase in resistance, the total oocyst output decreased, but the percentage of sporulation did not change significantly. We identified a total of 7511 peptides and 1282 proteins, and found 152 DEPs in the R10 strain versus the S strain, 426 DEPs in the R200 strain versus the S strain and 494 DEPs in the R200 strain versus the R10 strain. When compared with the S strain, 86 DEPs were found to have consistent trends in both resistant strains. The DEPs were primarily involved in ATP and GTP binding, invasion, and membrane components. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses of the DEPs suggested that they are involved in transcription and translation processes. Protein-protein interaction network analysis of the 86 DEPs showed that 10 proteins were hubs in the functional interaction network (≥ 8 edges) and five of them were ribosomal proteins.ConclusionsThe results of the present study indicate that the resistance mechanisms of E. tenella against EZL might be related to the transcriptional and translational processes, especially in the factors that inhibit the growth of parasites. The DEPs found in this study provide new insights into the resistance mechanisms of E. tenella against EZL. Further research on these potential targets holds promise for new chemotherapeutic approaches for controlling E. tenella infections.

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ITRAQ-based comparative proteomic analysis of cells infected with Eimeria tenella sporozoites.

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iTRAQ-based comparative proteomic analysis of cells infected with Eimeria tenella sporozoites.

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