Transfection of the protozoan parasite Perkinsus marinus

Fernández-Robledo, José A.; Lin, Zhuoer; Vasta, Gerardo R.

doi:10.1016/j.molbiopara.2007.09.007

Cited by 38 publications

(51 citation statements)

References 55 publications

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“…The trophozoites proliferate in phagosome-like structures, eventually lysing the host hemocytes, and causing systemic infection and death of the oyster, which by releasing large numbers of parasites into the water column spreads the infection to neighboring oysters (20, 35). Thus, the expression of CvGal2, as well as CvGal1 (22, 23), is associated with the cells and tissues that have been proposed as the entry sites for P. marinus trophozoites (20, 35, 37). Further, although CvGal2 is abundant in the hemocyte cytoplasm, it is also secreted into the extracellular space and binds to the hemocyte membrane glycocalyx.…”

Section: Resultsmentioning

confidence: 99%

“…Clonal cultures of P. marinus and P. chesapeaki strains both established in our laboratory [ P. marinus strain PRA 240 (37), P. chesapeaki (= P. andrewsi ) (14)], and obtained from ATCC were propagated as reported earlier (38). Cultures in log phase of growth were harvested and fixed with PFA as previously described (22, 23) for staining and flow cytometry analysis.…”

Section: Methodsmentioning

confidence: 99%

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Galectin CvGal2 from the Eastern Oyster (Crassostrea virginica) Displays Unique Specificity for ABH Blood Group Oligosaccharides and Differentially Recognizes Sympatric Perkinsus Species

et al. 2015

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Galectins are highly conserved lectins that are key to multiple biological functions, including pathogen recognition and regulation of immune responses. We previously reported that CvGal1, a galectin expressed in phagocytic cells (hemocytes) of the eastern oyster (Crassostrea virginica), is “hijacked” by the parasite Perkinsus marinus to enter the host, where it causes systemic infection and death. A screening of an oyster hemocyte cDNA library revealed a novel galectin, which we designated CvGal2, with four tandemly arrayed carbohydrate recognition domains (CRDs). A phylogentic analysis of the CvGal2 CRDs suggests close relationships with homologous CRDs from CvGal1. A glycan array analysis, however, revealed that unlike CvGal1 that preferentially binds to the blood group A tetrasaccharide, CvGal2 recognizes both blood group A and B tetrasaccharides and related structures, suggesting that CvGal2 has broader binding specificity. Further, SPR analysis demonstrated significant differences in the binding kinetics of CvGal1 and CvGal2, and structural modeling revealed substantial differences in their interactions with the oligosaccharide ligands. CvGal2 is homogeneously distributed in the hemocyte cytoplasm, is released to the extracellular space, and binds to the hemocyte surface. CvGal2 binds to P. marinus trophozoites in a dose-dependent and β-galactoside-specific manner. Strikingly, negligible binding of CvGal2 was observed for P. chesapeaki, a sympatric parasite species mostly prevalent in the clams Mya arenaria and Macoma balthica. The differential recognition of Perkinsus species by the oyster galectins is consistent with their relative prevalence in oyster and clam species, and supports their role in facilitating parasite entry and infectivity in a host-preferential manner.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Galectin CvGal2 from the Eastern Oyster (Crassostrea virginica) Displays Unique Specificity for ABH Blood Group Oligosaccharides and Differentially Recognizes Sympatric Perkinsus Species

et al. 2015

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“…Parasite (P. marinus) Trophozoites-Clonal cultures of P. marinus established in our laboratory (P. marinus strain PRA 240) (22) were propagated as reported earlier (23). Cultures in log phase of growth were harvested, washed three times in HBS, suspended at 4 ϫ 10 7 cells/ml in HBS, and treated with glutaraldehyde (10 mM) for 30 min on wet ice.…”

Section: Methodsmentioning

confidence: 99%

The Galectin CvGal1 from the Eastern Oyster (Crassostrea virginica) Binds to Blood Group A Oligosaccharides on the Hemocyte Surface*

Feng

Ghosh

Amin

et al. 2013

Journal of Biological Chemistry

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120

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Background:The carbohydrate specificity of the oyster galectin CvGal1 for endogenous and exogenous glycans was unresolved. Results: CvGal1 recognizes blood group A tetrasaccharides on oyster hemocytes, which are absent on the surface of the P. marinus parasite. Conclusion: Oyster hemocytes and P. marinus display structurally distinct ligands for CvGal1. Significance: Galectins may function as pattern recognition receptors by binding microbial glycans structurally different from endogenous ligands.

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“…This is especially relevant for diseases caused by apicomplexan parasites for which there are not effective vaccines [62]. Based on the large number of genes and pathways shared by Perkinsus and the apicomplexan [13], [14], and the availability of a transfection system [63], this approach has the potential to become an alternative for delivering vaccines against the apicomplexan parasites.…”

Section: Discussionmentioning

confidence: 99%

Humanized HLA-DR4 Mice Fed with the Protozoan Pathogen of Oysters Perkinsus Marinus (Dermo) Do Not Develop Noticeable Pathology but Elicit Systemic Immunity

et al. 2014

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Perkinsus marinus (Phylum Perkinsozoa) is a marine protozoan parasite responsible for “Dermo” disease in oysters, which has caused extensive damage to the shellfish industry and estuarine environment. The infection prevalence has been estimated in some areas to be as high as 100%, often causing death of infected oysters within 1–2 years post-infection. Human consumption of the parasites via infected oysters is thus likely to occur, but to our knowledge the effect of oral consumption of P. marinus has not been investigated in humans or other mammals. To address the question we used humanized mice expressing HLA-DR4 molecules and lacking expression of mouse MHC-class II molecules (DR4.EA0) in such a way that CD4 T cell responses are solely restricted by the human HLA-DR4 molecule. The DR4.EA0 mice did not develop diarrhea or any detectable pathology in the gastrointestinal tract or lungs following single or repeated feedings with live P. marinus parasites. Furthermore, lymphocyte populations in the gut associated lymphoid tissue and spleen were unaltered in the parasite-fed mice ruling out local or systemic inflammation. Notably, naïve DR4.EA0 mice had antibodies (IgM and IgG) reacting against P. marinus parasites whereas parasite specific T cell responses were undetectable. Feeding with P. marinus boosted the antibody responses and stimulated specific cellular (IFNγ) immunity to the oyster parasite. Our data indicate the ability of P. marinus parasites to induce systemic immunity in DR4.EA0 mice without causing noticeable pathology, and support rationale grounds for using genetically engineered P. marinus as a new oral vaccine platform to induce systemic immunity against infectious agents.

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Transfection of the protozoan parasite Perkinsus marinus

Cited by 38 publications

References 55 publications

Galectin CvGal2 from the Eastern Oyster (Crassostrea virginica) Displays Unique Specificity for ABH Blood Group Oligosaccharides and Differentially Recognizes Sympatric Perkinsus Species

Galectin CvGal2 from the Eastern Oyster (Crassostrea virginica) Displays Unique Specificity for ABH Blood Group Oligosaccharides and Differentially Recognizes Sympatric Perkinsus Species

The Galectin CvGal1 from the Eastern Oyster (Crassostrea virginica) Binds to Blood Group A Oligosaccharides on the Hemocyte Surface*

Humanized HLA-DR4 Mice Fed with the Protozoan Pathogen of Oysters Perkinsus Marinus (Dermo) Do Not Develop Noticeable Pathology but Elicit Systemic Immunity

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