Molecular cloning, sequencing, and expression of Eimeria tenella HSP70 partial gene

Bogado, Alexey Leon Gomel; Martins, Guilherme Felippelli; Sasse, João Pedro; Guimarães, José da Silva; Garcia, João Luís

doi:10.4238/gmr16018964

Cited by 5 publications

(4 citation statements)

References 25 publications

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“…HSP-70 plays an important role during sporulation and it facilitates stabilization of transmembrane proteins during host cell invasion (Bogado et al, 2017). In T. gondii , HSP-70 expression is increased as a response to stress by the host immune attack (Miller et al, 1999).…”

Section: Discussionmentioning

confidence: 99%

In vitro activity of selected natural products against Eimeria tenella sporozoites using Reproduction Inhibition Assay

Thabet,

Alzuheir,

Alnassan

et al. 2024

Preprint

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Eimeria tenella is the causative agent of cecal coccidiosis characterized by weight loss, hemorrhagic diarrhea, and high mortality rates. Research into herbal candidates to control the disease increased in the last decades by studying plant extracts with possible anticoccidial activity. As alternative to animal experiments, an in vitro reproduction inhibition assay (RIA) was previously designed to determine the sensitivity of E. tenella isolates against ionophores. In this study, RIA was used to test the anticoccidial activity of nutmeg oil, cinnamon oil, and glabridin. The concentration of nutmeg oil used in this study ranged between 1.1 μg/ml and139.1 μg/ml. Nutmeg oil exhibited a moderate in vitro inhibitory activity ranging from 35.5% to 49.5%. In contrast, no inhibitory effect was detected by incubating E. tenella sporozoites for 24 h with cinnamon oil at concentrations of 0.3 μg/ml to80.5 μg/ml. Glabridin (0.08 - 41.7 μg/ml) impaired sporozoites to replicate at a rate of14.1% to81.7% of inhibition. The calculated minimum concentrations of glabridin needed to inhibit parasite replication by 75%, 50%, and 30% (MIC , MIC , and MIC ) were 21.43μg/ml, 5.28μg/ml, and 0.96 μg/ml, respectively. Further studies to assess the in vitro efficacy of glabridin were performed by studying mRNA gene expression of stress provoker genes (HSP-70, NADPH, and EtPP5) after exposure of E. tenella sporozoites to glabridin at MIC for 0.5 h, 1 h, 2 h, and 4 h (time-dependent experiment). Moreover, a dose-dependent experiment was performed using glabridin at a concentration resembling MIC , MIC , and MIC for 24 h. In the time-dependent experiment, NADPH and EtPP5 were significantly ( p<0.05) overexpressed after 4 h of incubation with glabridin at a concentration of 21.43 μg/ml. The dose-dependent experiment exhibited a gradual overexpression in all studied genes which indicates stress imposed by glabridin on E. tenella sporozoites. In our hands, RIA was suitable toassess the strength of anticoccidial activity exhibited by the tested natural products. Such in vitro assays help to identify novel anticoccidial candidates such as herbal extracts.

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Section: Discussionmentioning

confidence: 99%

In vitro activity of selected natural products against Eimeria tenella sporozoites using Reproduction Inhibition Assay

Thabet,

Alzuheir,

Alnassan

et al. 2024

Preprint

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“…2 ) suggests that specific protein could be secreted via two or more secretory pathways during host-parasite interactions. The secretion of Hsp70 by Eimeria species is associated with sporozoite formation, pathogenicity, protective immunity and response to stress [ 5 , 58 , 59 ]. Hsp70 has also been reported among EV proteins from C. parvum, N. caninum , T. gondii , Leishmania and Trypanosoma species [ 7 ].…”

Section: Discussionmentioning

confidence: 99%

Eimeria falciformis secretes extracellular vesicles to modulate proinflammatory response during interaction with mouse intestinal epithelial cells

et al. 2022

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Background Protozoan parasite secretions can be triggered by various modified media and diverse physicochemical stressors. Equally, host-parasite interactions are known to co-opt the exchange and secretion of soluble biochemical components. Analysis of Eimeria falciformis sporozoite secretions in response to interaction with mouse intestinal epithelial cells (MIECs) may reveal parasite secretory motifs, protein composition and inflammatory activities of E. falciformis extracellular vesicles (EVs). Methods Eimeria falciformis sporozoites were allowed to interact with inactivated MIECs. Parasite secretions were separated into EV and vesicle-free (VF) fractions by discontinuous centrifugation and ultracentrifugation. Secreted EVs were purified in an iodixanol density gradient medium and the protein composition of both EV and VF fractions were analyzed by liquid chromatoraphy-tandem mass spectroscopy. The inflammatory activities of E. falciformis sporozoite EV on MIECs were then investigated. Results During the interaction of E. falciformis sporozoites with inactivated MIECs, the parasite secreted VF and vesicle-bound molecules. Eimeria falciformis vesicles are typical pathogenic protozoan EVs with a mean diameter of 264 ± 2 nm, and enclosed heat shock protein (Hsp) 70 as classical EV marker. Refractile body-associated aspartyl proteinase (or eimepsin), GAP45 and aminopeptidase were the main components of E. falciformis sporozoite EVs, while VF proteins include Hsp90, actin, Vps54 and kinases, among others. Proteomic data revealed that E. falciformis EV and VF proteins are aggregates of bioactive, antigenic and immunogenic molecules which act in concert for E. falciformis sporozoite motility, pathogenesis and survival. Moreover, in MIECs, E. falciformis EVs induced upregulation of gene expression and secretion of IL-1β, IL-6, IL-17, IL-18, MCP1 as well as pyroptosis-dependent caspase 11 and NLRP6 inflammasomes with the concomitant secretion of lactate dehydrogenase. Conclusions Eimeria falciformis sporozoite interaction with MIECs triggered the secretion of immunogenic and antigenic proteins. In addition, E. falciformis sporozoite EVs constitute parasite-associated molecular pattern that induced inflammatory response and cell death. This study offers additional insight in the secretion and protein composition of E. falciformis secretomes as well as the proinflammatory functions of E. falciformis sporozoite EVs. Graphical Abstract

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“…However, heterogeneity or nucleotide diversity of protein genes could vary in different isolates [ 60 ]. Other prominent protein genes that have been mapped are hsp90 [ 57 ], hsp70 [ 61 ] as well as protease genes in which > 40 are already identified in E. tenella genome [ 62 ]. Although eimerian developmental stages share many transcriptional and translational products [ 63 ], each stage possibly has varying threshold of gene expression [ 64 ] and translational profiles [ 55 ].…”

Section: A Glimpse Into Protein Gene Profilesmentioning

confidence: 99%

“…Despite this, Hsp70 plays an important role in the formation of sporocysts and sporozoites [ 61 ] (Table 1 ). A dose-dependent inhibition of Hsp70 by quercetin inhibited the formation of syneptonema complex and haploidy in E. tenella sporozoite suggesting that Hsp70 could act as sentinel for assembly and disassembly of other proteins during developmental transition [ 141 ].…”

Section: Cytoplasmic Proteinsmentioning

confidence: 99%

Eimeria proteins: order amidst disorder

Olajide

Yang

et al. 2022

Parasites Vectors

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Apicomplexans are important pathogens that cause severe infections in humans and animals. The biology and pathogeneses of these parasites have shown that proteins are intrinsically modulated during developmental transitions, physiological processes and disease progression. Also, proteins are integral components of parasite structural elements and organelles. Among apicomplexan parasites, Eimeria species are an important disease aetiology for economically important animals wherein identification and characterisation of proteins have been long-winded. Nonetheless, this review seeks to give a comprehensive overview of constitutively expressed Eimeria proteins. These molecules are discussed across developmental stages, organelles and sub-cellular components vis-à-vis their biological functions. In addition, hindsight and suggestions are offered with intention to summarise the existing trend of eimerian protein characterisation and to provide a baseline for future studies. Graphical Abstract

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Molecular cloning, sequencing, and expression of Eimeria tenella HSP70 partial gene

Cited by 5 publications

References 25 publications

In vitro activity of selected natural products against Eimeria tenella sporozoites using Reproduction Inhibition Assay

In vitro activity of selected natural products against Eimeria tenella sporozoites using Reproduction Inhibition Assay

Eimeria falciformis secretes extracellular vesicles to modulate proinflammatory response during interaction with mouse intestinal epithelial cells

Eimeria proteins: order amidst disorder

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