Identification and characterisation of a dense granuleassociated protein in Neospora caninum tachyzoites

Hemphill, Andrew; Gajendran, N; Sonda, Sabrina; Fuchs, Nicole; Gottstein, Bruno; Hentrich, Brigitte; Jenkins, Mark C.

doi:10.1016/s0020-7519(97)00193-8

Cited by 35 publications

(23 citation statements)

References 26 publications

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“…LR White embedding and on-section labeling of N. caninum-infected Vero cell cultures were performed as previously described (11,16,18). Sections were labeled with affinity-purified NcMIC4 antibodies diluted 1:2 in PBS-0.1% BSA, and bound antibodies were detected by using goat anti-rabbit antibody conjugated to 10-nm gold particles (Amersham, Zürich, Switzerland).…”

Section: Methodsmentioning

confidence: 99%

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Identification and Characterization of a Neospora caninum Microneme-Associated Protein (NcMIC4) That Exhibits Unique Lactose-Binding Properties

et al. 2004

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Microneme proteins have been shown to play an important role in the early phase of host cell adhesion, by mediating the contact between the parasite and host cell surface receptors. In this study we have identified and characterized a lectin-like protein of Neospora caninum tachyzoites which was purified by ␣-lactose-agarose affinity chromatography. Upon separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, this lactose-binding protein migrated at 70 and 55 kDa under reducing and nonreducing conditions, respectively. Immunofluorescence and immunogold electron microscopy with affinity-purified antibodies showed that the protein was associated with the tachyzoite micronemes. Mass spectrometry analyses and expressed sequence tag database mining revealed that this protein is a member of the Neospora microneme protein family; the protein was named NcMIC4 (N. caninum microneme protein 4). Upon two-dimensional gel electrophoresis, NcMIC4 separated into seven distinct isoforms. Incubation of extracellular parasites at 37°C resulted in the secretion of NcMIC4 into the medium as a soluble protein, and the secreted protein exhibited a slightly reduced

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Section: Methodsmentioning

confidence: 99%

“…7A). As a control the same fractions were stained with anti-NcMIC1 (positive control [22]) and anti-NcGRA7 antibodies (negative control [18]) (Fig. 7B and C, respectively).…”

Section: Ncmic4-host Cell Interactionmentioning

confidence: 99%

Identification and Characterization of a Neospora caninum Microneme-Associated Protein (NcMIC4) That Exhibits Unique Lactose-Binding Properties

et al. 2004

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“…This was also found earlier in a comparative ultrastructural study by Speer et al (1999), which reported that in cells infected with T. gondii VEG strain tachyzoites, several host cell mitochondria were closely associated with the PVM, whereas only few host cell mitochondria were found in the vicinity to the PVM surrounding N. caninum tachyzoites. In addition, secreted NcGRA7 was found to be associated with the PVM (Hemphill et al 1998).…”

Section: Host Cell Modulation and Parasite-host Cell Crosstalkmentioning

confidence: 98%

“…On the molecular level, the initial low affinity host-parasite contact is mediated, at least in part, through the two major surface antigens of N. caninum tachyzoites, NcSAG1 and NcSRS2 (Howe et al 1998, Sonda et al 1998. In vitro studies have shown that both polyclonal and monoclonal antibodies directed against these two surface antigens inhibit host cell adhesion and invasion , Nishikawa et al 2000.…”

Section: Neospora Caninum Surface Components Implicated In Adhesion Amentioning

confidence: 99%

“…Of these, T. gondii ROP2 has been reported to insert into the vacuole membrane and mediate anchoring to host cell mitochondria (Sinai and Joiner 2001). Following invasion, the lumen of the vacuoles of both parasites as well as its membrane is extensively modified through secretory products, most likely originating from rhoptries and dense granules (Cesbron-Delauw 1994, Hemphill et al 1998. Since in T. gondii-infected cells, the PVM actively recruits host cell mitochondria and ER, protocols have been developed for the enrichment of this organellar membrane complex (Sinai et al 1997).…”

Section: Host Cell Modulation and Parasite-host Cell Crosstalkmentioning

confidence: 99%

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Neospora caninum and neosporosis — recent achievements in host and parasite cell biology and treatment

Hemphill

Gottstein

2006

Acta Parasitologica

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Neospora caninum is an apicomplexan parasite, which owes its importance to the fact that it represents the major infectious cause of bovine abortion worldwide. Its life cycle is comprised of three distinct stages: Tachyzoites, representing the proliferative and disease-causing stage, bradyzoites, representing a slowly replicating, tissue cyst-forming stage, and sporozoites, which represent the end product of a sexual process taking place within the intestinal tissue of the final canine host. Tachyzoites are capable of infecting a large variety of host cells in vitro and in vivo, while bradyzoites have been found mainly within the central nervous system. In order to survive, proliferate, and proceed in its life cycle, N. caninum has evolved some amazing features. First, the parasite profits immensely from its ability to interact with, and invade, a large number of host cell types. Secondly, N. caninum exploits its capability to respond to alterations in living conditions by converting into another stage (tachyzoite-to-bradyzoite or vice versa). Thirdly, this parasite has evolved mechanisms that modulate its host cells according to its own requirements, and these must, especially in the case of the bradyzoite stage, involve mechanisms that ensure long term survival of not only the parasite but also of the host cell. These three key events (host cell invasion -stage conversion -host cell modulation) represent potential targets for intervention. In order to elucidate the molecular and cellular bases of these important features of N. caninum, cell culture-based approaches and laboratory animal models are extensively exploited. In this review, we will summarize the present knowledge and achievements related to host cell and parasite cell biology.

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Function of Neospora caninum dense granule protein 7 in innate immunity in mice

Wang

Song

et al. 2020

Parasitol Res

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Identification and characterisation of a dense granuleassociated protein in Neospora caninum tachyzoites

Cited by 35 publications

References 26 publications

Identification and Characterization of a Neospora caninum Microneme-Associated Protein (NcMIC4) That Exhibits Unique Lactose-Binding Properties

Identification and Characterization of a Neospora caninum Microneme-Associated Protein (NcMIC4) That Exhibits Unique Lactose-Binding Properties

Neospora caninum and neosporosis — recent achievements in host and parasite cell biology and treatment

Function of Neospora caninum dense granule protein 7 in innate immunity in mice

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