The production of toxic oxygen metabolites by hemocytes of different snail species

Dikkeboom, Ronald; Knapp, Wil P.W. van der; Bovenkamp, Wim van den; Tijnagel, Jolanda M.G.H.; Bayne, Christopher J.

doi:10.1016/0145-305x(88)90068-7

Cited by 54 publications

(13 citation statements)

References 27 publications

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“…SOD is known to catalyse the decomposition of superoxide anion (0;) into O2 and H 2 0 2 (McCord & Fridovich 1969). The CL inhibition observed when SOD was used at 80 ,pg ml-l (50'0) (as opposed to no effect at 10 pg ml-') is consistent with data concerning luminolenhanced CL for human neutrophils (De Chatelet et al 19821, murine polymorphonuclear granulocytes and macrophages (Muller-Peddinghaus 1984, Vincendeau et al 1988) and gastropod hemocytes (Dikkeboom et al 1987(Dikkeboom et al , 1988. NaN3 inhibits myeloperoxidase (MPO) (Nakagawara et al 1981) and catalase, and has a quencher effect on singlet oxygen ('O2) (Rosen & Klebanoff 1977).…”

Section: Discussionsupporting

confidence: 73%

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Chemiluminescence analysis of the activity of Pecten maximua hemocytes stimulated with zymosan and host-specific Rickettsiales-like organisms

Gall¹,

Bachère²,

Mialhe³

1991

Dis. Aquat. Org.

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ABSTRACT. During a study of Rickettsiales-like organisms (RLO) in Pecten maximus gill, histological observation of free RLO in hemolymph led us to consider interactions between the pathogen and the hemocytes. Firstly, the production of oxygen radicals by hemocytes was established using a luminolenhanced chemiluminescence method with zymosan as stimulant. The effects of several inh~bitors of oxidative metabolism were investigated. The internalization of RLO into hemocytes was demonstrated by electron microscopy but was not related to chemiluminescent activity. The possible involvement of RLO acid phosphatase as the inhibitor of respiratory burst was assessed.

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Section: Discussionsupporting

confidence: 73%

“…By refemng to vertebrate phagocytic cells (Roos 1980), this methodology has been developed for several gastropod species -Lyrnnaea stagnalis (Dikkeboom et al 1987), Planorborius corneus, Helix aspersa (Dikkeboom et al 1988) and Biomphalaria glabrata (Shozawa 1986) -and recently for oysters (Larson et al 1989, Bachgre et al 1991). …”

Section: Introductionmentioning

confidence: 99%

Chemiluminescence analysis of the activity of Pecten maximua hemocytes stimulated with zymosan and host-specific Rickettsiales-like organisms

Gall¹,

Bachère²,

Mialhe³

1991

Dis. Aquat. Org.

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“…As also demonstrated in several gastropod species (Dikkeboom et al 1988b), w e showed by the CL technique that oyster hemocytes display metabolic events associated with the phagocytosis of zymosan particles. In practice, the study was not a simple extrapolation of gastropod studies, since the establishment of the CL technique required the definition of several controlled experimental conditions, which had to be adapted to oyster hemocytes.…”

Section: Discussionmentioning

confidence: 94%

Lumhol-dependent chemiluminescence by hemocytes of two marine bivalves, Ostrea edulis and Crassostrea gigas

Bachère¹,

Hervio²,

Mialhe³

1991

Dis. Aquat. Org.

107

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Microbicidal phagocytic function and oxidative metabolism of the hemocytes of 2 oyster species, Ostrea edulis and Crassostrea gigas, were investigated using a luminol-enhanced chemiluminescence (CL) technique. First, experimental parameters adapted to marine bivalve hemocytes were established on hemolymph pools of C. gigas in order to obtain qualitatively and quantitatively homogeneous samples and so to perform statistically comparable assays. The use of Modified Alsever Solution (MAS) allowed hemocytes to b e kept non-aggregated and in a non-stimulated state until starting the assays. A number of aliquots of 2 X lo5 hemocytes with a n MAS final concentration of 3.5 % showed high chemiluminescent response after stimulation by zymosan particles A partic1e:hemocyte ratio of 80:1 gave optimal CL activity. Activity was inhibited by cytochalasin B, a phagocyt~c inhibitor, and by sodlum azide, the latter indicating the involvement of oxygen products. Having thus defined suitable parameter values, the CL protocol was then applled to 0. edulis for qualitative a n d quantitative analyses of respiratory burst capacity at the species and individual levels. Results suggested inter-and intra-specific variability of CL responses. Under the same experimental conditions, 0. eduhs hemocytes generally displayed higher CL activities than C. gigas. Moreover, clear individual vanabllity was demonstrated. The same experimental number of hemocytes showed great differences in CL responses between individuals suggesting a posslble correlation with hemogram characteristics such a s cell type percentages.

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“…Interestingly, it has been suggested that the means by which resistant individuals of the host snail kill sporocysts include the production of reactive oxygen species (29). Despite this sensitivity, sporocyst growth and embryogenesis could continue under normoxic conditions (air) when 2-mercaptoethanol and Bge cells were both present.…”

Section: Discussionmentioning

confidence: 99%

Continuous in vitro propagation and differentiation of cultures of the intramolluscan stages of the human parasite Schistosoma mansoni

Ivanchenko

Lerner²,

McCormick³

et al. 1999

Proc. Natl. Acad. Sci. U.S.A.

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The metazoan parasitic blood f lukes, Schistosoma spp., infect over 200 million people worldwide and cause extensive human morbidity and mortality. Research strategies for development of anti-schistosomal agents are impeded by the organism's complex molluscan-mammalian life cycle, which limits experimental approaches and availability of material. We derived long-term continuously proliferative cultures of Schistosoma mansoni sporocysts capable of generating cercariae in vitro. Cultured organisms retained the ability to parasitize the host, and they exhibited developmental regulation of candidate stage-specific genes in the host-free culture system. Evidence for expression of a reverse transcriptase also was found in the cultured organisms, pointing to this activity as a possible mechanistic contributor to the dynamic relationship between the parasite and its hosts. Continuous in vitro propagation of the asexual sporocyst stage allows isolation of clonally derived parasite populations and provides a means to study schistosomal molecular genetics, metabolism, and evasion of host defenses.Schistosoma mansoni, Schistosoma haematobium, and Schistosoma japonicum create widespread disease in tropical developing countries (1). These parasites thwart attempts to produce practical and effective vaccines, and pharmacological approaches are problematic (2). Schistosome eggs are laid by females paired with males and residing in the vasculature of parasitized vertebrate hosts. To be successfully infective, eggs must penetrate the intestinal wall and pass in feces to fresh water. From these released eggs hatch free-swimming miracidia, the first larval form. Successful miracidia encounter and penetrate the appropriate species of snail and develop into sporocysts, the second larval from. In the following weeks, these primary sporocysts asexually generate first-generation and second-generation daughter sporocysts in the snail. From the later-generation sporocysts develop the third larval form, free-swimming cercaria. To be successfully infective in the human or other vertebrate host, cercaria must encounter and penetrate the skin of the host and undergo further changes to form schistosomula. These move through several further developmental stages, ultimately leading to sexual pairing in the veins of the parasitized host and egg production.Approaches to elucidate the mechanisms schistosomes employ to escape host immunity, as well as molecular genetic and pharmacological investigations, have been limited by the lack of an in vitro system for the continuous proliferative culture of the parasite in the absence of the host. Recently, Yoshino and colleagues (3, 4) have reported significant success in limited culture of the intramolluscan stages of S. mansoni and S.japonicum, but these techniques do not allow extended culture of proliferating sporocysts.Here we describe long-term cultures of S. mansoni in vitro and the development from the snail-infective miracidial stage, through generations of mother and daughter sporocysts, t...

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The production of toxic oxygen metabolites by hemocytes of different snail species

Cited by 54 publications

References 27 publications

Chemiluminescence analysis of the activity of Pecten maximua hemocytes stimulated with zymosan and host-specific Rickettsiales-like organisms

Chemiluminescence analysis of the activity of Pecten maximua hemocytes stimulated with zymosan and host-specific Rickettsiales-like organisms

Lumhol-dependent chemiluminescence by hemocytes of two marine bivalves, Ostrea edulis and Crassostrea gigas

Continuous in vitro propagation and differentiation of cultures of the intramolluscan stages of the human parasite Schistosoma mansoni

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