Regulation of oyster (Crassostrea virginica) hemocyte motility by the intracellular parasite Perkinsus marinus: A possible mechanism for host infection

Lau, Yuk-Ting; Gambino, Laura; Santos, Bianca Silva; Espinosa, Emmanuelle Pales; Allam, Bassem

doi:10.1016/j.fsi.2018.04.019

Cited by 14 publications

(8 citation statements)

References 46 publications

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“…In North America, P. marinus and Perkinsus chesapeaki can coexist in the same bivalve host (McLaughlin and Faisal, 1998;Coss et al, 2001a,b;Pecher et al, 2008;Reece et al, 2008;Arzul et al, 2012). In the oysters, the parasite is taken up by hemocytes and uses them as a vehicle for migration into other host tissues (Lau et al, 2018a;Schott et al, 2019;Yadavalli et al, 2020). Studies based on intracellular structures and phylogeny suggest P. marinus as a close relative to the apicomplexan, a lineage leading to intracellular parasitism having shared genomic and physiological affinities (Matsuzaki et al, 2008;Joseph et al, 2010;Bachvaroff et al, 2011;Fernández Robledo et al, 2011;Van Voorhis et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

CRISPR/Cas9 Ribonucleoprotein-Based Genome Editing Methodology in the Marine Protozoan Parasite Perkinsus marinus

Yadavalli

Umeda

Waugh

et al. 2021

Front. Bioeng. Biotechnol.

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Perkinsus marinus (Perkinsozoa), a close relative of apicomplexans, is an osmotrophic facultative intracellular marine protozoan parasite responsible for “Dermo” disease in oysters and clams. Although there is no clinical evidence of this parasite infecting humans, HLA-DR40 transgenic mice studies strongly suggest the parasite as a natural adjuvant in oral vaccines. P. marinus is being developed as a heterologous gene expression platform for pathogens of medical and veterinary relevance and a novel platform for delivering vaccines. We previously reported the transient expression of two rodent malaria genes Plasmodium berghei HAP2 and MSP8. In this study, we optimized the original electroporation-based protocol to establish a stable heterologous expression method. Using 20 μg of pPmMOE[MOE1]:GFP and 25.0 × 106P. marinus cells resulted in 98% GFP-positive cells. Furthermore, using the optimized protocol, we report for the first time the successful knock-in of GFP at the C-terminus of the PmMOE1 using ribonucleoprotein (RNP)-based CRISPR/Cas9 gene editing methodology. The GFP was expressed 18 h post-transfection, and expression was observed for 8 months post-transfection, making it a robust and stable knock-in system.

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Section: Introductionmentioning

confidence: 99%

CRISPR/Cas9 Ribonucleoprotein-Based Genome Editing Methodology in the Marine Protozoan Parasite Perkinsus marinus

Yadavalli

Umeda

Waugh

et al. 2021

Front. Bioeng. Biotechnol.

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“…So far, there are many reports on other tetraspanins in nematode, mollusk, arthropod, and ascidian (24)(25)(26)(27), which could offer useful leads to the investigation of CD63 subfamily for invertebrates. For instance, invertebrate tetraspanins could be significantly induced by the stimulations of various pathogens (28)(29)(30)(31)(32)(33), and they could function as mediators of innate immune response (34,35). Tetraspanin D76 from insect (Manduca sexta) was found to play an important role in cell-mediated immune responses by influencing the encapsulation and clearance of bacteria (35).…”

Section: Introductionmentioning

confidence: 99%

A CD63 Homolog Specially Recruited to the Fungi-Contained Phagosomes Is Involved in the Cellular Immune Response of Oyster Crassostrea gigas

et al. 2020

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Cluster of differentiation 63 (CD63), a four-transmembrane glycoprotein in the subfamily of tetraspanin, has been widely recognized as a gateway from the infection of foreign invaders to the immune defense of hosts. Its role in Pacific oyster Crassostrea gigas is, however, yet to be discovered. This work makes contributions by identifying Cg CD63H, a CD63 homolog with four transmembrane domains and one conservative CCG motif, and establishing its role as a receptor that participates in immune recognition and hemocyte phagocytosis. The presence of Cg CD63H messenger RNA (mRNA) in hepatopancreas, labial palps, gill, and hemocytes is confirmed. The expression level of mRNA in hemocytes is found significantly ( p < 0.01) upregulated after the injection of Vibrio splendidus . Cg CD63H protein, typically distributed over the plasma membrane of oyster hemocytes, is recruited to the Yarrowia lipolytica -containing phagosomes after the stimulation of Y. lipolytica . The recombinant Cg CD63H protein expresses binding capacity to glucan (GLU), peptidoglycan (PGN), and lipopolysaccharide (LPS) in the presence of lyophilized hemolymph. The phagocytic rate of hemocytes toward V. splendidus and Y. lipolytica is significantly inhibited ( p < 0.01) after incubation with anti-CgCD63H antibody. Our work further suggests that Cg CD63H functions as a receptor involved in the immune recognition and hemocyte phagocytosis against invading pathogen, which can be a marker candidate for the hemocyte typing in C. gigas .

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“…We should also note that to date, all the Perkinsus spp. have been described as parasites of mollusks and they have evolved to adhere to, enter the oyster, and survive inside the defense cells of mollusks [35,36,39] (reviewed in [40]). Consequently, the protozoan detected in the oysters can only come from infected oysters.…”

Section: Discussionmentioning

confidence: 99%

A qPCR-Based Survey of Haplosporidium nelsoni and Perkinsus spp. in the Eastern Oyster, Crassostrea virginica in Maine, USA

et al. 2020

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Eastern oyster (Crassostrea virginica) aquaculture is increasingly playing a significant role in the state of Maine’s (USA) coastal economy. Here, we conducted a qPCR-based survey for Haplosporidium nelsoni, Perkinsus marinus, and Perkinsus chesapeaki in C. virginica (n = 1440) from six Maine sites during the summer–fall of 2016 and 2017. In the absence of reported die-offs, our results indicated the continued presence of the three protozoan parasites in the six sites. The highest H. nelsoni qPCR-prevalence corresponded to Jack’s Point and Prentiss Island ( x ¯ = 40 and 48% respectively), both located in the Damariscotta River Estuary. Jack’s Point, Prentiss Island, New Meadows River, and Weskeag River recorded the highest qPCR-prevalence for P. marinus (32–39%). While the P. marinus qPCR-prevalence differed slightly for the years 2016 and 2017, P. chesapeaki qPCR-prevalence in 2016 was markedly lower than 2017 (<20% at all sites versus >60% at all sites for each of the years, respectively). Mean qPCR-prevalence values for P. chesapeaki over the two-year study were ≥40% for samples from Jack’s Point (49%), Prentiss Island (44%), and New Meadows River (40%). This study highlights that large and sustained surveys for parasitic diseases are fundamental for decision making toward the management of the shellfish aquaculture industry, especially for having a baseline in the case that die-offs occur.

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Regulation of oyster (Crassostrea virginica) hemocyte motility by the intracellular parasite Perkinsus marinus: A possible mechanism for host infection

Cited by 14 publications

References 46 publications

CRISPR/Cas9 Ribonucleoprotein-Based Genome Editing Methodology in the Marine Protozoan Parasite Perkinsus marinus

CRISPR/Cas9 Ribonucleoprotein-Based Genome Editing Methodology in the Marine Protozoan Parasite Perkinsus marinus

A CD63 Homolog Specially Recruited to the Fungi-Contained Phagosomes Is Involved in the Cellular Immune Response of Oyster Crassostrea gigas

A qPCR-Based Survey of Haplosporidium nelsoni and Perkinsus spp. in the Eastern Oyster, Crassostrea virginica in Maine, USA

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