The Molecular Characterization and Immunity Identification of Microneme 3 of <i>Eimeria acervulina</i>

Zhang, Zhenchao; Liu, Xinchao; Yang, XinChao; Liu, Lianrui; Wang, Shuai; Lu, Mingmin; Ehsan, Muhammad; Gadahi, Javaid Ali; Song, Xiaoling; Xu, Lixin; Ren, Yan; Li, Xiangrui

doi:10.1111/jeu.12318

Cited by 17 publications

(14 citation statements)

References 49 publications

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“…Jenkins et al [ 66 ] showed the immune-mapped protein 1 could be detected in the sporozoites. Zhang et al [ 31 ] found EaMIC3 on the apical tip of E. acervulina sporozoites. Our findings suggest that EmSAG is expressed in the sporozoite and merozoite stages of E. maxima , and might play an important role in the host invasion mechanism.…”

Section: Discussionmentioning

confidence: 99%

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Molecular characterisation and the protective immunity evaluation of Eimeria maxima surface antigen gene

et al. 2018

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BackgroundCoccidiosis is recognised as a major parasitic disease in chickens. Eimeria maxima is considered as a highly immunoprotective species within the Eimeria spp. family that infects chickens. In the present research, the surface antigen gene of E. maxima (EmSAG) was cloned, and the ability of EmSAG to stimulate protection against E. maxima was evaluated.MethodsProkaryotic and eukaryotic plasmids expressing EmSAG were constructed. The EmSAG transcription and expression in vivo was performed based on the RT-PCR and immunoblot analysis. The expression of EmSAG in sporozoites and merozoites was detected through immunofluorescence analyses. The immune protection was assessed based on challenge experiments. Flow cytometry assays were used to determine the T cell subpopulations. The serum antibody and cytokine levels were evaluated by ELISA.ResultsThe open reading frame (ORF) of EmSAG gene contained 645 bp encoding 214 amino acid residues. The immunoblot and RT-PCR analyses indicated that the EmSAG gene were transcribed and expressed in vivo. The EmSAG proteins were expressed in sporozoite and merozoite stages of E. maxima by the immunofluorescence assay. Challenge experiments showed that both pVAX1-SAG and the recombinant EmSAG (rEmSAG) proteins were successful in alleviating jejunal lesions, decreasing loss of body weight and the oocyst ratio. Additionally, these experiments possessed anticoccidial indices (ACI) of more than 170. Higher percentages of CD4+ and CD8+ T cells were detected in both EmSAG-inoculated birds than those of the negative control groups (P < 0.05). The EmSAG-specific antibody concentrations of both the rEmSAG and pVAX1-EmSAG groups were much higher than those of the negative controls (P < 0.05). Higher concentrations of IL-4, IFN-γ, TGF-β1 and IL-17 were observed more in both the rEmSAG protein and pVAX1-SAG inoculated groups than those of negative controls (P < 0.05).ConclusionsOur findings suggest that EmSAG is capable of eliciting a moderate immune protection and could be used as an effective vaccine candidate against E. maxima.

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Section: Discussionmentioning

confidence: 99%

“…The counts of T cells in the treatment groups were evaluated by flow cytometry analysis as previously described [ 30 , 31 ]. Spleens were extracted from 5 chickens of each group at pre-, first-, and second-vaccination days.…”

Section: Methodsmentioning

confidence: 99%

Molecular characterisation and the protective immunity evaluation of Eimeria maxima surface antigen gene

et al. 2018

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“…Chapman and Shirley (102) used a modified version of the former protocol and assigned cut-offs for drug sensitivity; >160 = sensitive, partially resistant = 120-160 and <120 = resistant. Some vaccine studies have used ACI as a method to evaluate vaccine efficacy and assigned different cut-off scores for efficacy with some variation between studies; an ACI of <160 is generally considered to be ineffective (103)(104)(105)(106)(107). However, whether this adapted scoring method is overly simplistic to evaluate the efficacy and usefulness of vaccines in the field has yet to be determined.…”

Section: Composite Measures Of Efficacymentioning

confidence: 99%

Poultry Coccidiosis: Design and Interpretation of Vaccine Studies

et al. 2020

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Eimeria infection impacts upon chicken welfare and economic productivity of the poultry sector. Live coccidiosis vaccines for chickens have been available for almost 70 years, but the requirement to formulate blends of oocysts from multiple Eimeria species makes vaccine production costly and logistically demanding. A multivalent vaccine that does not require chickens for its production and can induce protection against multiple Eimeria species is highly desirable. However, despite the identification and testing of many vaccine candidate antigens, no recombinant coccidiosis vaccine has been developed commercially. Currently, assessment of vaccine efficacy against Eimeria, and the disease coccidiosis, can be done only through in vivo vaccination and challenge experiments but the design of such studies has been highly variable. Lack of a "standard" protocol for assessing vaccine efficacy makes comparative evaluations very difficult, complicating vaccine development, and validation. The formulation and schedule of vaccination, the breed of chicken and choice of husbandry system, the species, strain, magnitude, and timing of delivery of the parasite challenge, and the parameters used to assess vaccine efficacy all influence the outcomes of experimental trials. In natural Eimeria infections, the induction of strong cell mediated immune responses are central to the development of protective immunity against coccidiosis. Antibodies are generally regarded to be of lesser importance. Unfortunately, there are no specific immunological assays that can accurately predict how well a vaccine will protect against coccidiosis (i.e., no "correlates of protection"). Thus, experimental vaccine studies rely on assessing a variety of post-challenge parameters, including assessment of pathognomonic lesions, measurements of parasite replication such as oocyst output or quantification of Eimeria genomes, and/or measurements of productivity such as body weight gain and feed conversion rates. Understanding immune responses to primary and secondary infection can inform on the most appropriate immunological assays. The discovery of new antigens for different Eimeria species and the development of new methods of vaccine antigen delivery necessitates a more considered approach to assessment of novel vaccines with robust, repeatable study design. Careful consideration of performance and welfare factors that are genuinely relevant to chicken producers and vaccine manufacturers is essential.

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“…Interference of c-Cbl expression can promote apoptosis by upregulating the expression of Fas and DR4/5 ( 33 , 34 ). Also, c-Cbl can degrade caspase-8 by ubiquitination and then inhibit cell apoptosis in gastric cancer ( 35 ). This result suggests that c-Cbl can inhibit host cell apoptosis via substrate protein ubiquitination.…”

Section: Discussionmentioning

confidence: 99%

Eimeria acervulina Microneme Protein 3 Inhibits Apoptosis of the Chicken Duodenal Epithelial Cell by Targeting the Casitas B-Lineage Lymphoma Protein

et al. 2021

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Eimeria acervulina (E. acervulina) causes coccidiosis in poultry which persists as economic pain worldwide. Most damage to the intestinal mucosa results from apoptosis of the infected intestinal epithelial cells. The Microneme protein 3 (MIC3) protein is a key virulence factor in some parasites involved in host cell apoptosis inhibition. Here, we studied whether and how MIC3 affects the apoptosis in E. acervulina infected chicken duodenal epithelial cells. Through flow cytometry (FCM), we found that the presence of merozoites and the overexpression of MIC3 significantly decreased apoptosis and the activity of caspase-3 in chicken duodenal epithelial cells at 4, 6, and 8 h post merozoite infection (P < 0.01). Silencing the Casitas B-lineage lymphoma (CBL) protein, a host receptor for MIC3 with shRNA was shown to promote apoptosis in the chicken duodenal epithelial cells. The early apoptotic rate of host cells in the lentiviral-MIC3 group was significantly lower than that in the lentiviral-MIC3 + shRNA CBL group at 4 h after MIC3 expression (P < 0.01), and it was moderately decreased in the lentiviral-MIC3 + shRNA CBL group compared with that in the shRNA CBL group. Our data indicated that MIC3 inhibited early apoptosis of E. acervulina infected chicken duodenal epithelial cells by targeting host receptor-CBL protein. These findings unveiled one of the mechanisms of how intracellular parasites affect the apoptosis of infected host cells, which provided a deeper understanding of their pathogenesis.

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The Molecular Characterization and Immunity Identification of Microneme 3 of Eimeria acervulina

Cited by 17 publications

References 49 publications

Molecular characterisation and the protective immunity evaluation of Eimeria maxima surface antigen gene

Molecular characterisation and the protective immunity evaluation of Eimeria maxima surface antigen gene

Poultry Coccidiosis: Design and Interpretation of Vaccine Studies

Eimeria acervulina Microneme Protein 3 Inhibits Apoptosis of the Chicken Duodenal Epithelial Cell by Targeting the Casitas B-Lineage Lymphoma Protein

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