A method for the isolation of schistosome eggs and 

miracidia free of contaminating host tissues

Dalton, John P.; Day, Sharon R.; Drew, Angela F.; Brindley, Paul J.

doi:10.1017/s0031182097001091

Cited by 142 publications

(136 citation statements)

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“…Worms were prepared as described previously 32 and snap frozen in liquid nitrogen. S. haematobium eggs were isolated from the livers from infected hamsters 33 and washed extensively in saline. All samples were frozen at −80 °C.…”

Section: Methodsmentioning

confidence: 99%

Whole-genome sequence of Schistosoma haematobium

et al. 2012

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Schistosomiasis is a neglected tropical disease caused by blood flukes (genus Schistosoma; schistosomes) and affecting 200 million people worldwide 1 . No vaccines are available, and treatment relies on one drug, praziquantel 2 . Schistosoma haematobium has come into the spotlight as a major cause of urogenital disease, as an agent linked to bladder cancer 1,3 and as a predisposing factor for HIV/AIDS 4,5 . The parasite is transmitted to humans from freshwater snails 1 . Worms dwell in blood vessels and release eggs that become embedded in the bladder wall to elicit chronic immune-mediated disease 6 and induce squamous cell carcinoma 7 . Here we sequenced the 385-Mb genome of S. haematobium using Illumina-based technology at 74-fold coverage and compared it to sequences from related parasites 8,9 . We included genome annotation based on function, gene ontology, networking and pathway mapping. This genome now provides an unprecedented resource for many fundamental research areas and shows great promise for the design of new disease interventions.

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Section: Methodsmentioning

confidence: 99%

Whole-genome sequence of Schistosoma haematobium

et al. 2012

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“…For this, ~500 schistosomules in ~ 200 μl Modified Eagle's Medium were injected into the thigh muscle of mice using a 22G syringe, as described [18]. Sixty days later, the mice were euthanized, adult worms recovered by portal vein perfusion, gross pathology of livers examined, and schistosome eggs recovered from the livers by digestion with bacterial collagenase [21].…”

Section: Electroporation Of Schistosomulesmentioning

confidence: 99%

RNA interference of Schistosoma mansoni cathepsin D, the apical enzyme of the hemoglobin proteolysis cascade

Morales

Rinaldi

Gobert

et al. 2008

Molecular and Biochemical Parasitology

119

111

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The aspartic protease cathepsin D (Clan AA, Family A1) is expressed in the schistosome gut where it plays an apical role in the digestion of hemoglobin released from ingested erythrocytes. In this report, RNA interference approaches were employed to investigate the effects of knockdown of schistosome cathepsin D. Cultured schistosomules of Schistosoma mansoni were exposed by square wave electroporation to double stranded RNA (dsRNA) specific for cDNA encoding S. mansoni cathepsin D. RNAi mediated reductions in transcript levels led to phenotypic changes including significant growth retardation in vitro and suppression of aspartic protease enzyme activity. In addition, black-pigmented heme, the end point by-product of normal hemoglobin proteolysis that accumulates in the schistosome gut, was not apparent within the guts of the treated schistosomules. Rather, their guts appeared to be red in color, rather than black, apparently indicating the presence of intact rather than digested host hemoglobin. These phenotypic effects were apparent when either of two forms of dsRNA, a long form spanning the entire target transcript or a short form specific for the 3'-region were employed. Off-target effects were not apparent in transcript levels of the gut-localized cysteine protease cathepsin B1. Finally, cathepsin D may be an essential enzyme in the mammal-parasitic stages of schistosomes because schistosomules treated ex vivo with dsRNA did not survive to maturity after transfer into Balb/c mice. These and earlier findings suggest that, given its essential function in parasite nutrition, schistosome cathepsin D could be developed as a target for novel anti-schistosomal interventions.

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“…Six-to 12-wkold females were used as bone marrow donors for DC culture, for in vivo immunizations, or as splenocyte donors for CD4 T cell purification for use in some in vitro experiments. SEA was prepared from isolated schistosome eggs, as previously described (33,34). Care was used to prevent endotoxin contamination of the Ag preparation, and using a timed gel endotoxin detection kit (Sigma, St. Louis, MO), we found the SEA to be contamination free.…”

Section: Animals and Reagentsmentioning

confidence: 99%

CD8− Dendritic Cell Activation Status Plays an Integral Role in Influencing Th2 Response Development

MacDonald

Straw

Bauman

et al. 2001

The Journal of Immunology

255

292

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Whether dendritic cells (DC) play a passive or active role in Th2 response induction is poorly understood. In this study, we show that CD8− DC pulsed with Th2-polarizing Ag (soluble egg Ag (SEA)) from Schistosoma mansoni potently stimulate Th2 responses in vivo and in vitro while failing to undergo a conventional maturation process. Thus, in contrast to DC pulsed with the Th1 response inducing Ag Propionebacterium acnes, SEA-exposed DC exhibit a phenotype that is most similar to that of immature DC, failing to up-regulate expression of CD40, CD54, CD80, CD86, or OX40L; producing no detectable IL-4, IL-10, or IL-12; and displaying only a minor increase in MHC class II expression. Importantly, in vitro derived DC exposed to SEA were phenotypically similar to CD8− DC isolated from active S. mansoni infection. By discriminating between different types of pathogen and responding appropriately, CD8− DC play a major role in the decision process to mount either a Th1 or Th2 response.

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A method for the isolation of schistosome eggs and miracidia free of contaminating host tissues

Cited by 142 publications

References 0 publications

Whole-genome sequence of Schistosoma haematobium

Whole-genome sequence of Schistosoma haematobium

RNA interference of Schistosoma mansoni cathepsin D, the apical enzyme of the hemoglobin proteolysis cascade

CD8− Dendritic Cell Activation Status Plays an Integral Role in Influencing Th2 Response Development

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