Protective immunity induced by Eimeria common antigen 14–3-3 against Eimeria tenella, Eimeria acervulina and Eimeria maxima

Tian

et al. 2021

Vet Res

Self Cite

Rhomboid-like proteases (ROMs) are considered as new candidate antigens for developing new-generation vaccines due to their important role involved in the invasion of apicomplexan protozoa. In prior works, we obtained a ROM2 sequence of Eimeria maxima (EmROM2). This study was conducted to evaluate the immunogenicity and protective efficacy of EmROM2 recombinant protein (rEmROM2) and EmROM2 DNA (pVAX1-EmROM2) against infection by Eimeria maxima (E. maxima). Firstly, Western blot assay was conducted to analyze the immunogenicity of rEmROM2. The result showed that rEmROM2 was recognized by chicken anti-E. maxima serum. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot assay revealed apparent transcription and expression of EmROM2 at the injection site. qRT-PCR (quantitative real-time PCR), flow cytometry and indirect ELISA indicated that vaccination with rEmROM2 or EmROM2 DNA significantly upregulated the transcription level of cytokines (IFN-γ, IL-2, IL-4, IL-10, IL-17, TGF-β and TNF SF15), the proportion of CD8+ and CD4+ T lymphocytes and serum IgG antibody response. Ultimately, a vaccination-challenge trial was performed to evaluate the protective efficacy of rEmROM2 and pVAX1-EmROM2 against E. maxima. The result revealed that vaccination with rEmROM2 or pVAX1-EmROM2 significantly alleviated enteric lesions, weight loss, and reduced oocyst output caused by challenge infection of E. maxima, and provided anticoccidial index (ACI) of more than 160, indicating partial protection against E. maxima. In summary, vaccination with rEmROM2 or pVAX1-EmROM2 activated notable humoral and cell-mediated immunity and provided partial protection against E. maxima. These results demonstrated that EmROM2 protein and DNA are promising vaccine candidates against E. maxima infection.

Section: Discussionmentioning

confidence: 99%

Rhomboid protein 2 of Eimeria maxima provided partial protection against infection by homologous species

Tian

et al. 2021

Vet Res

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“…Liu et al [29] identified immunodominant proteins of three Eimeria species (E. tenella, E. maxima and E. acervulina), in which five groups of the protein orthologs were detected as common antigens. The E. acervulina 14-3-3 antigen was shown to induce significant immune responses and provided protection against E. tenella, E. acervulina and E. maxima infection [35]. The results of the present study added more putative, common, immunoreactive proteins to the list of crossprotective antigens, which should aid in facilitating the development of multivalent vaccines against co-infections by pathogenic Eimeria species.…”

Section: Discussionmentioning

confidence: 73%

Immunoproteomic and mass spectrometric analysis of Eimeria acervulina antigens recognized by antisera from chickens infected with E. acervulina, E. tenella or E. necatrix

Liu

Tuo

et al. 2020

Parasites Vectors

Background: Coccidiosis is caused by Eimeria spp. and can result in severe economic losses to the global poultry industry. Due to anticoccidial drug resistance rapidly developing in the parasites and drug residues in poultry products, efficacious and safe alternative coccidia control measures are needed. The objective of the present study was to identify common protective antigens which may be used as vaccine candidates in the development of subunit, multivalent, cross-protective vaccines against most of the economically important Eimeria species.Methods: Whole sporozoite proteins of Eimeria acervulina were prepared and analyzed by 2-dimensional gel electrophoresis (2-DE) followed by western blotting using immune sera specific to E. tenella, E. acervulina, or E. necatrix. The protein spots detected by all three immune sera were then excised from the preparative gel and protein ID was performed by MALDI-TOF-MS/MS. Results:Approximately 620 E. acervulina sporozoite protein spots were demonstrated by 2-DE with silver staining, among which 23 protein spots were recognized by immune sera specific to all three Eimeria species. The results showed that 21 putative E. acervulina proteins were identified, which include proteins with known enzymatic properties, and those which are involved in protein translation, transport and trafficking, and ribosomal biogenesis and functions. There is one protein which may be involved in transcription and one heat-shock protein. Two proteins contain predicted domains, but with no apparent functions known. There were 2 protein spots which had no detectable proteins. None of the proteins has a predicted signal peptide or a transmembrane domain; however, 6 of the 21 putative proteins were predicted to be potentially secretory through the non-classical pathway. Conclusions:Our study identified a diverse group of antigens immunologically common to all three Eimeria species, none of which was previously characterized and tested as a vaccine candidate. Further research on immunogenicity and cross-protective potential of these individual proteins as vaccine candidates will aid the development of vaccines against the most common and pathogenic Eimeria spp.

“…In recent years, couples of antigens of E. maxima were tested as candidate antigens for subunit or DNA vaccines and showed promising protective e cacy, such as gam56 and gam82 [29], EmMIC2 and EmMIC7 [30,31] and some Eimeria common antigens (e.g. GAPDH and 14-3-3) [8,32]. Since ROMs are involved in the invasion of apicomplexan protozoa, they were considered as a new candidate antigen for developing new-generation vaccine [13,14].…”

Section: Discussionmentioning

confidence: 99%

Rhomboid Protein 2 of Eimeria Maxima Provided Partial Protection Against Infection by Homologous Species

Tian

et al. 2020

Preprint

Self Cite

Background: Rhomboid-like proteases (ROMs) are considered as a new candidate antigen for developing new-generation vaccine due to their important role involved in the invasion of apicomplexan protozoa. In prior works, we obtained a ROM2 sequence of Eimeria maxima (EmROM2) which is the homologous gene with ROM2 of Toxoplasma gondii. This study was conducted to evaluate the immunogenicity and protective efficacy of EmROM2 recombinant protein (rEmROM2) and EmROM2 DNA (pVAX1-EmROM2) against infection by Eimeria maxima (E. maxima).Methods: Western blot assay was conducted to analyze the immunogenicity of rEmROM2. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot assay were performed to determine the transcription and expression of pVAX1-EmROM2 recombinant plasmid. EmROM2-induced changes in transcriptional level of cytokines, T lymphocytes subsets and specific serum IgG antibody were detected through qPCR (quantitative real-time PCR), flow cytometry and indirect ELISA, respectively. Ultimately, a vaccination-challenge trial was performed to evaluate the protective efficacy of rEmROM2 and pVAX1-EmROM2 against infection with E. maxima. Results: The purified rEmROM2 was recognized with chicken anti-E. maxima serum. After vaccination with pVAX1-EmROM2, apparent transcription and translation of EmROM2 were observed in the vaccinated chickens. Vaccination with rEmROM2 and EmROM2 DNA significantly upregulated the proportion of CD8+ and CD4+ T lymphocytes, the transcription level of cytokines (IFN-γ, IL-2, IL-4, IL-10, IL-17, TGF-β and TNF SF15) and serum IgG antibody response. Meanwhile, the vaccination significantly alleviated enteric lesions, weight loss, and reduced oocyst output caused by challenge infection of E. maxima, and provided anticoccidial index (ACI) of more than 160, indicating partial protection against E. maxima.Conclusions: Vaccination with rEmROM2 and pVAX1-EmROM2 activated notable humoral and cell-mediated immunity and provided partial protection against infection by E. maxima. These results demonstrated that EmROM2 protein and DNA are promising vaccine candidates against E. maxima infection.