Proteins mediating the Neospora caninum-host cell interaction as targets for vaccination

Hemphill, Andrew; Debache, Karim; Monney, Thierry; Schorer, Michelle; Guionaud, Christophe; Alaeddine, Ferial; Mueller, Norbert; Mueller, Joachim D.

doi:10.2741/e593

Cited by 37 publications

(37 citation statements)

References 96 publications

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“…Those most relevant for farm animals are Babesia , Besnoitia , Cryptosporidium , Eimeria , Neospora , Sarcocystis , Theileria , and Toxoplasma , all causing diseases of great socio-economic impact worldwide ( Müller and Hemphill, 2013 ). Neospora caninum is phylogenetically closely related to Toxoplasma gondii , but distinct from Toxoplasma with regard to several biological features including the life cycle, host range and pathogenicity ( Hemphill et al., 2006, 2013 ). Canids, namely dogs, wolves, dingoes and coyotes, represent definitive hosts of N. caninum .…”

Section: Introductionmentioning

confidence: 99%

Buparvaquone is active against Neospora caninum in vitro and in experimentally infected mice

Müller

Aguado‐Martínez

Manser

et al. 2015

International Journal for Parasitology: Drugs and Drug Resistan

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Section: Introductionmentioning

confidence: 99%

Buparvaquone is active against Neospora caninum in vitro and in experimentally infected mice

Müller

Aguado‐Martínez

Manser

et al. 2015

International Journal for Parasitology: Drugs and Drug Resistan

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“…The lytic cycle has been extensively studied in the closely related parasite Toxoplasma gondii [ 4 , 5 ], but only scarcerly investigated in N. caninum . However, preliminary studies showed that the molecular mechanisms that control this process are highly conserved among T. gondii and N. caninum [ 6 ].…”

Section: Introductionmentioning

confidence: 99%

The tandemly repeated NTPase (NTPDase) from Neospora caninum is a canonical dense granule protein whose RNA expression, protein secretion and phosphorylation coincides with the tachyzoite egress

et al. 2016

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BackgroundNTPases (also NTPDases) are enzymes with apyrase activity. They are widely distributed among eukaryotes, and also among members of the family Sarcocystidae. In Toxoplasma gondii, the TgNTPase accumulates in the dense granules, and has been commonly associated with the strain virulence. In the closely related Neospora caninum, the NcNTPase lacks nucleoside diphosphate hydrolase activity and appears to be more abundant in virulent isolates, indicating that it may contribute to the pathogenicity of neosporosis. However, so far no additional information on NcNTPase has been provided.MethodsHerein, the NcNTPase coding sequences were analysed by different in silico and de novo sequencing approaches. A comparative analysis of NcNTPase and NcGRA7 in terms of protein dynamics, secretion, phosphorylation, and mRNA expression profiles during the tachyzoite lytic cycle was also carried out. Moreover, NcNTPase immunolocalization was analysed by confocal microscopy techniques over a set number of time-points.ResultsWe describe the presence of three different loci containing three copies of the NcNTPase within the Nc-Liv genome, and report the existence of up to four different NcNTPase alleles in Nc-Liv. We also provide evidence for the occurrence of diverse protein species of the NcNTPase by two-dimensional gel electrophoresis. Both NcNTPase and NcGRA7 were similarly up-regulated and secreted during the egress and/or early invasion phases, and were phosphorylated. However, its secretion was not affected by the addition of calcium modulators such as A23187 and ethanol. NcNTPase and NcGRA7 localized in dense granules and parasitophorous vacuole membrane throughout the lytic cycle, although differed in their inmunolocalization during early invasion and egress.ConclusionsThe present study reveals the complexity of the NcNTPase loci in N. caninum. We hypothesize that the expression of different isoforms of the NcNTPase protein could contribute to parasite virulence. Our findings showed regulation of expression, secretion and phosphorylation of NcNTPase suggesting a potential role for progression through the tachyzoites lytic cycle.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-016-1620-4) contains supplementary material, which is available to authorized users.

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“…Because invasion is considered a requirement for pathogen survival and replication within the host, the identification of secreted proteins (secretome) involved in invasion may be useful to reveal interesting targets for therapeutic intervention [ 8 , 11 ].…”

Section: Introductionmentioning

confidence: 99%

Unravelling the Neospora caninum secretome through the secreted fraction (ESA) and quantification of the discharged tachyzoite using high-resolution mass spectrometry-based proteomics

et al. 2013

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BackgroundThe apicomplexan parasite Neospora caninum causes neosporosis, a disease that leads to abortion or stillbirth in cattle, generating an economic impact on the dairy and beef cattle trade. As an obligatory intracellular parasite, N. caninum needs to invade the host cell in an active manner to survive. The increase in parasite cytosolic Ca2+ upon contact with the host cell mediates critical events, including the exocytosis of phylum-specific secretory organelles and the activation of the parasite invasion motor. Because invasion is considered a requirement for pathogen survival and replication within the host, the identification of secreted proteins (secretome) involved in invasion may be useful to reveal interesting targets for therapeutic intervention.MethodsTo chart the currently missing N. caninum secretome, we employed mass spectrometry-based proteomics to identify proteins present in the N. caninum tachyzoite using two different approaches. The first approach was identifying the proteins present in the tachyzoite-secreted fraction (ESA). The second approach was determining the relative quantification through peptide stable isotope labelling of the tachyzoites submitted to an ethanol secretion stimulus (discharged tachyzoite), expecting to identify the secreted proteins among the down-regulated group.ResultsAs a result, 615 proteins were identified at ESA and 2,011 proteins quantified at the discharged tachyzoite. We have analysed the connection between the secreted and the down-regulated proteins and searched for putative regulators of the secretion process among the up-regulated proteins. An interaction network was built by computational prediction involving the up- and down-regulated proteins. The mass spectrometry proteomics data have been deposited to the ProteomeXchange with identifier PXD000424.ConclusionsThe comparison between the protein abundances in ESA and their measure in the discharged tachyzoite allowed for a more precise identification of the most likely secreted proteins. Information from the network interaction and up-regulated proteins was important to recognise key proteins potentially involved in the metabolic regulation of secretion. Our results may be helpful to guide the selection of targets to be investigated against Neospora caninum and other Apicomplexan organisms.

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Proteins mediating the Neospora caninum-host cell interaction as targets for vaccination

Cited by 37 publications

References 96 publications

Buparvaquone is active against Neospora caninum in vitro and in experimentally infected mice

Buparvaquone is active against Neospora caninum in vitro and in experimentally infected mice

The tandemly repeated NTPase (NTPDase) from Neospora caninum is a canonical dense granule protein whose RNA expression, protein secretion and phosphorylation coincides with the tachyzoite egress

Unravelling the Neospora caninum secretome through the secreted fraction (ESA) and quantification of the discharged tachyzoite using high-resolution mass spectrometry-based proteomics

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