The genome of the protozoan parasite Cystoisospora suis and a reverse vaccinology approach to identify vaccine candidates

Palmieri, Nicola; Shrestha, Aruna; Ruttkowski, Bärbel; Beck, Tomas; Vogl, Claus; Tomley, Fiona M.; Blake, Damer P.; Joachim, Anja

doi:10.1016/j.ijpara.2016.11.007

Cited by 31 publications

(21 citation statements)

References 78 publications

(95 reference statements)

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“…The SAGs protein of Eimeria tenella is capable of inducing an immune response against coccidiosis in chickens [ 12 ]. Therefore, surface antigens and cell adhesion proteins have been suggested as promising vaccine candidates against parasitic infections [ 13 , 14 ].…”

Section: Introductionmentioning

confidence: 99%

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: Introductionmentioning

confidence: 99%

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

et al. 2018

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“…Toxoplasma gondii is a member of the Coccidia, a subclass of apicomplexans known to form environmentally resistant oocysts, which, if sporulated and ingested via food or water, may provoke host infection ( Fritz et al 2012 ; Lindsay et al 1997 ). Besides T. gondii , Coccidia comprise other single-celled obligate parasites such as Eimeria , Isospora , Neospora , Cyclospora , Sarcocystis , and Hammondia , which usually infect vertebrates ( Blazejewski 2015 ; Liu et al 2016 ; Palmieri et al 2017 ). Similar to other Coccidia, T. gondii has a complex life cycle involving three different infectious stages: Tachyzoites, bradyzoites, and sporozoites ( Dubey et al 1998 ).…”

Section: Introductionmentioning

confidence: 99%

Genomic and Proteomic Evidence for the Presence of a Peroxisome in the Apicomplexan Parasite Toxoplasma gondii and Other Coccidia

Moog

Przyborski

Maier

2017

Genome Biology and Evolution

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Apicomplexans are successful parasites responsible for severe human diseases including malaria, toxoplasmosis, and cryptosporidiosis. For many years, it has been discussed whether these parasites are in possession of peroxisomes, highly variable eukaryotic organelles usually involved in fatty acid degradation and cellular detoxification. Conflicting experimental data has been published. With the age of genomics, ever more high quality apicomplexan genomes have become available, that now allow a new assessment of the dispute. Here, we provide bioinformatic evidence for the presence of peroxisomes in Toxoplasma gondii and other coccidians. For these organisms, we have identified a complete set of peroxins, probably responsible for peroxisome biogenesis, division, and protein import. Moreover, via a global screening for peroxisomal targeting signals, we were able to show that a complete set of fatty acid β-oxidation enzymes is equipped with either PTS1 or PTS2 sequences, most likely mediating transport of these factors to putative peroxisomes in all investigated Coccidia. Our results further imply a life cycle stage-specific presence of peroxisomes in T. gondii and suggest several independent losses of peroxisomes during the evolution of apicomplexan parasites.

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“…gondii and Neospora caninum, signifying the close relationship of C. suis with these two species [42], and more than 36% with Eimeria necatrix (Table 1).…”

Section: Genes Linked To Sexual Stagesmentioning

confidence: 97%

Characterization of Cystoisospora suis sexual stages in vitro

Feix¹,

Cruz‐Bustos²,

Ruttkowski³

et al. 2019

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Background: The porcine coccidium Cystoisospora suis is characterized by a complex life cycle during which asexual multiplication is followed by sexual development with two morphologically distinct cell types, the micro- and macrogametes. Genes related to the sexual stages and cell cycle progression were previously identified in related Apicomplexa. Here, the sexual stages of C. suis were characterized in vitro morphologically and for temporal expression changes of the mentioned genes to gain insight into this poorly known phase of coccidian development.Methods: Sexual stages of C. suis (micro- and macrogametes) developing in vitro in porcine intestinal epithelial cells were examined by light and electron microscopy. The transcriptional levels of genes related to merozoite multiplication and sexual development were evaluated by quantitative real-time PCR at different time points of cultivation. Dynein light chain type 1 and male gamete fusion factor HAP2 expression is presumably restricted to microgametes. Tyrosin-rich proteins are a part of the wall-forming bodies and oocyst wall proteins. The RAD51/Dmc1 protein plays a role during the cell division process, and Nima-related protein kinases regulate the cell cycle and are associated with male or female Plasmodium gametocytes. Transcription levels were compared for parasites in culture supernatants at 6–9 doc (day of cultivation) and intracellular parasites at 6–15 doc.Results: Sexual stage of C. suis could be found during 8–11 doc in vitro. Microgametes had a spherical body (3.0–5.0 µm) and two flagella (10.8–12.3 µm). Macrogametes were spherical with a diameter of 11.5–13.0 µm. Merozoite gene transcription peaked on 10 doc and then declined. Genes related to the sexual stages and cell cycle showed an upregulation with a peak on 13 doc, after which they declined. Conclusions: The present work linked gene expression changes to the detailed morphological description of C. suis sexual development in vitro, including fertilization, meiosis and oocyst formation in vitro. Following this process at the cellular and molecular level will elucidate details on potential bottlenecks of development of C. suis (and coccidian parasites in general) which could be exploited as novel targets for control.

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The genome of the protozoan parasite Cystoisospora suis and a reverse vaccinology approach to identify vaccine candidates

Abstract: Graphical abstract

Cited by 31 publications

References 78 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

Genomic and Proteomic Evidence for the Presence of a Peroxisome in the Apicomplexan Parasite Toxoplasma gondii and Other Coccidia

Characterization of Cystoisospora suis sexual stages in vitro

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