Partial protective of chickens against Eimeria tenella challenge with recombinant EtMIC-1 antigen

Qi, Nanshan; Wang, Y. Y.; Liao, Shenquan; Wu, Caiyan; Lv, Minna; Li, J.; Tong, Zongxi; Sun, Mingfei

doi:10.1007/s00436-013-3389-0

Cited by 20 publications

(10 citation statements)

References 25 publications

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“…This needs to be further investigated. Qi et al (2013) found that EtMIC1 induced significant level of the IFN-c and IL-2 transcript profiles at first immunization, and reached the peak at third immunization, and Zhang et al (2014b) reported that EaMIC5 could provocate high level of the IL-4 and sCD8. However, in this research, we found that EaMIC3 induced significant level of the IFN-c, IL-10, IL-17, and TGF-b at first immunization, and reached the peak at second immunization.…”

Section: Discussionmentioning

confidence: 97%

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

Zhang

Liu

Yang

et al. 2016

J Eukaryotic Microbiology

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The gene of Eimeria acervulina microneme protein 3 (EaMIC3) was cloned and characterized. According to the conserved sequence, the 3'- and 5'-ends of EaMIC3 were amplified by the rapid amplification of cDNA ends (RACE). The full length cDNA of this gene was obtained by overlapping the sequences of 3'- and 5'-extremities and amplification by reverse transcription PCR. The sequence analysis revealed that the opening reading frame (ORF) of EaMIC3 was 2,607 bp and encoded a protein of 868 amino acids with 93.04 kDa. Western blotting assay showed that the recombinant protein was successfully recognized by the sera of chickens experimentally infected with E. acervulina, whereas the native protein in the somatic extract of sporozoites was as well detected by sera from rats immunized with the recombinant protein of EaMIC3. Immunofluorescence analysis indicated that EaMIC3 was expressed in the sporozoites and merozoites stages of E. acervulina. Animal challenge experiments demonstrated that the recombinant protein of EaMIC3 could significantly increase the average body weight gains, decrease the mean lesion scores and the oocyst outputs of the immunized chickens and presented anticoccidial index (ACI) more than 165. Moreover, EaMIC3 protein produced significantly high level of IgG antibody, IFN-γ, IL-10, IL-17 TGF-β, CD4 , and CD8 .

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Section: Discussionmentioning

confidence: 97%

“…Qi et al. () found that EtMIC1 induced significant level of the IFN‐γ and IL‐2 transcript profiles at first immunization, and reached the peak at third immunization, and Zhang et al. () reported that EaMIC5 could provocate high level of the IL‐4 and sCD8.…”

Section: Discussionmentioning

confidence: 99%

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

Zhang

Liu

Yang

et al. 2016

J Eukaryotic Microbiology

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“…But we found EaMIC5 could incite significant level of sCD8 while the level sCD4 could not be promoted 10 days post booster. Moreover, Qi et al [42] found that EtMIC1 induced significant level of the IFN-γ transcript profiles at first immunization (at days 14), and reached the peak at third immunization (at days 28). All of the above results indicated that different antigens might induce different cytokine and CD4 + or CD8 + cells at different time points.…”

Section: Discussionmentioning

confidence: 99%

Identification and Molecular Characterization of Microneme 5 of Eimeria acervulina

Zhang

Huang

et al. 2014

PLoS ONE

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In the present study, the microneme 5 gene of Eimeria acervulina (E. acervulina) (EaMIC5) was cloned and characterized. Specific primers for the rapid amplification of cDNA ends (RACE) were designed based on the expressed sequence tag (EST, GenBank Accession No. EH386430.1) to amplify the 3′- and 5′-ends of EaMIC5. The full length cDNA of this gene was obtained by overlapping the sequences of 3′- and 5′-extremities and amplification by reverse transcription PCR. Sequence analysis revealed that the open reading frame (ORF) of EaMIC5 was 336 bp and encoded a protein of 111 amino acids with 12.18 kDa. The ORF was inserted into pET-32a (+) to produce recombinant EaMIC5. Using western blotting assay, the recombinant protein was successfully recognized by the sera of chicks experimentally infected with E. acervulina, while the native protein in the somatic extract of sporozoites was as well detected by sera from rats immunized with the recombinant protein of EaMIC5. Immunofluorescence analysis using antibody against recombinant protein EaMIC5 indicated that this protein was expressed in the sporozoites and merozoites stages of E. acervulina. Animal challenge experiments demonstrated that the recombinant protein of EaMIC5 could significantly increase the average body weight gains, decrease the mean lesion scores and the oocyst outputs of the immunized chickens, and presented anti-coccidial index (ACI) more than 160. All the above results suggested that the EaMIC5 was a novel E. acervulina antigen and could be an effective candidate for the development of a new vaccine against this parasite.

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“…in fowl. Meanwhile, both MIC1 and MIC3 proteins of E. tenella have been implicated in the process of invasion of the host cells and the merogony of parasites [ 44 , 46 , 47 ]. Therefore, both EsMIC1 and EsMIC3 may be the potential diagnostic and vaccine candidate antigens against hepatic coccidiosis.…”

Section: Discussionmentioning

confidence: 99%

Expression Analysis and Serodiagnostic Potential of Microneme Proteins 1 and 3 in Eimeria stiedai

Wei

Shen

Xiao

et al. 2020

Genes

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Eimeria stiedai is an apicomplexan protozoan parasite that invades the liver and bile duct epithelial cells in rabbits and causes severe hepatic coccidiosis, resulting in significant economic losses in the domestic rabbit industry. Hepatic coccidiosis lacks the typical clinical symptoms and there is a lack of effective premortem tools to timely diagnose this disease. Therefore, in the present study we cloned and expressed the two microneme proteins i.e., microneme protein 1 (EsMIC1) and microneme protein 3 (EsMIC3) from E. stiedai and used them as recombinant antigens to develop a serodiagnostic method for an effective diagnosis of hepatic coccidiosis. The cDNAs encoding EsMIC1 and EsMIC3 were cloned and the mRNA expression levels of these two genes at different developmental stages of E. stiedai were determined by quantitative real-time PCR analysis (qRT-PCR). The immunoreactivity of recombinant EsMIC1 (rEsMIC1) and EsMIC3 (rEsMIC3) proteins were detected by Western blotting, and indirect enzyme-linked immunosorbent assays (ELISAs) based on these two recombinant antigens were established to evaluate their serodiagnostic potential. Our results showed that the proteins encoded by the ORFs of EsMIC1 (711 bp) and EsMIC3 (891 bp) were approximately 25.89 and 32.39 kDa in predicted molecular weight, respectively. Both EsMIC1 and EsMIC3 showed the highest mRNA expression levels in the merozoites stage of E. stiedai. Western blotting analysis revealed that both recombinant proteins were recognized by E. stiedai positive sera, and the indirect ELISAs using rEsMIC1 and rEsMIC3 were developed based on their good immunoreactivity, with 100% (48/48) sensitivity and 97.9% (47/48) specificity for rEsMIC1 with 100% (48/48) sensitivity and 100% (48/48) specificity for rEsMIC3, respectively. Moreover, rEsMIC1- and rEsMIC3-based indirect ELISA were able to detect corresponding antibodies in sera at days 6, 8, and 10 post E. stiedai infection, with the highest positive diagnostic rate (62.5% (30/48) for rEsMIC1 and 66.7% (32/48) for rEsMIC3) observed at day 10 post infection. Therefore, both EsMIC1 and EsMIC3 can be used as potential serodiagnostic candidate antigens for hepatic coccidiosis caused by E. stiedai.

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Partial protective of chickens against Eimeria tenella challenge with recombinant EtMIC-1 antigen

Cited by 20 publications

References 25 publications

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

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

Identification and Molecular Characterization of Microneme 5 of Eimeria acervulina

Expression Analysis and Serodiagnostic Potential of Microneme Proteins 1 and 3 in Eimeria stiedai

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