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      iTRAQ-based comparative proteomic analysis of cells infected with Eimeria tenella sporozoites Translated title: Analyse protéomique comparative basée sur iTRAQ de cellules infectées par les sporozoïtes d’ Eimeria tenella

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          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.

          Translated abstract

          Eimeria tenella est un parasite intracellulaire obligatoire qui envahit activement les cellules épithéliales cæcales des poulets. Quand E. tenella infecte une cellule, l’hôte produit un changement correspondant pour traiter les dommages causés par cette infection. À ce jour, nos connaissances sur le mécanisme de réponse de la cellule hôte à l’infection à E. tenella sont très limitées, tant au niveau moléculaire que cellulaire. Dans cette étude, des marqueurs isobares pour la quantification relative et absolue (iTRAQ) couplés à la LC-MS / MS ont été utilisés pour cribler les protéines exprimées de manière différentielle (PED) dans des cellules BHK-21 infectées par des sporozoïtes de E. tenella, 24 h après l’infection. Au total, 6139 protéines distinctes non redondantes ont été identifiées, et 195 d’entre elles présentaient un taux de variation de ≥ 1,3 ou ≤ 0,7 et un p < 0,05, dont 151 protéines régulées positivement et 44 protéines régulées négativement. La fiabilité des données protéomiques a ensuite été validée avec qPCR et Western Blot. L’enrichissement par l’ontologie des gènes a indiqué que les PED régulés positivement étaient principalement impliquées dans la liaison et l’activité catalytique, alors que les PED régulées négativement étaient celles de l’activité catalytique et des régulateurs de la fonction moléculaire. De plus, l’analyse de la voie de KEGG a montré que les PED participaient aux voies de signalisation de PI3K-Akt, de la chémokine, de Ras, de Wnt, de p53, et autres, et que les PED régulées positivement et négativement étaient liées respectivement aux voies de surveillance du ribosome et de l’ARNm, respectivement. Les données de cette étude fournissent une base importante pour une analyse plus poussée des interactions entre les cellules hôtes d’ E. tenella.

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          Most cited references 36

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          Translation control: bridging the gap between genomics and proteomics?

          mRNA profiling enables the expression levels of thousands of transcripts in a cell to be monitored simultaneously. Nevertheless, analyses in yeast and mammalian cells have demonstrated that mRNA levels alone are unreliable indicators of the corresponding protein abundances. This discrepancy between mRNA and protein levels argues for the relevance of additional control mechanisms besides transcription. As translational control is a major mechanism regulating gene expression, the use of translated mRNA in profiling experiments might depict the proteome more closely than does the use of total mRNA. This would combine the technical potential of genomics with the physiological relevance of proteomics.
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            Chasing the golden egg: vaccination against poultry coccidiosis.

            Eimeria species, of the Phylum Apicomplexa, cause the disease coccidiosis in poultry, resulting in severe economic losses every year. Transmission of the disease is via the faecal-oral route, and is facilitated by intensive rearing conditions in the poultry industry. Additionally, Eimeria has developed drug resistance against most anticoccidials used today, which, along with the public demand for chemical free meat, has lead to the requirement for an effective vaccine strategy. This review focuses on the history and current status of anticoccidial vaccines, and our work in developing the transmission-blocking vaccine, CoxAbic (Netanya, Israel). The vaccine is composed of affinity-purified antigens from the wall-forming bodies of macrogametocytes of Eimeria maxima, which are proteolytically processed and cross-linked via tyrosine residues to form the environmentally resistant oocyst wall. The vaccine is delivered via maternal immunization, where vaccination of laying hens leads to protection of broiler offspring. It has been extensively tested for efficacy and safety in field trials conducted in five countries and involving over 60 million offspring chickens from immunized hens and is currently the only subunit vaccine against any protozoan parasite to reach the marketplace.
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              Activation of Wnt/beta-catenin pathway mediates growth and survival in B-cell progenitor acute lymphoblastic leukaemia.

              This study investigated the response of acute lymphoblastic leukaemia (ALL) cells to Wnt proteins. Accumulation of beta-catenin was measured by Western blotting and immunofluorescence microscopy. Reverse transcription polymerase chain reaction (RT-PCR) analysis of B-cell progenitor acute lymphoblastic leukaemia (ALL) cells revealed expression of Wnt genes, including WNT2B in 33%, WNT5A in 42%, WNT10B in 58% and WNT16B in 25% of cases. The Wnt receptors, (Frizzled) FZD7 and FZD8 were also expressed in most cases while FZD3, FZD4 and FZD9 were occasionally detected. Stimulation of ALL cells with Wnt-3a activated canonical Wnt signalling with increased expression and nuclear translocation of beta-catenin. This resulted in a 1.7- to 5.3-fold increase in cell proliferation, which was associated with enhanced cell cycle entry. A significant increase in the survival of ALL cells under conditions of serum deprivation was also observed. Microarray analysis and quantitative RT-PCR revealed that activation of the Wnt/beta-catenin pathway led to altered expression of genes involved in cell cycle regulation and apoptosis in normal and leukaemic B-cell progenitors. Our results demonstrate that Wnt-3a provides proliferative and survival cues in ALL cells. This data suggests that targeting the Wnt signalling pathway may be a useful therapeutic strategy in ALL.
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                Author and article information

                Journal
                Parasite
                Parasite
                parasite
                Parasite
                EDP Sciences
                1252-607X
                1776-1042
                2019
                20 February 2019
                : 26
                : ( publisher-idID: parasite/2019/01 )
                Affiliations
                Key Laboratory of Animal Parasitology of Ministry of Agriculture, Shanghai Veterinary Research Institute Shanghai 200241 PR China
                Author notes
                Article
                parasite180108 10.1051/parasite/2019009
                10.1051/parasite/2019009
                6383524
                30789155
                © Z. Zhao et al., published by EDP Sciences, 2019

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                Page count
                Figures: 5, Tables: 2, Equations: 0, References: 38, Pages: 10
                Categories
                Research Article

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