An in vitro assay utilising parasitic larval Haemonchus contortus to detect resistance to closantel and other anthelmintics

Rothwell, J.; Sangster, Nicholas C.

doi:10.1016/0020-7519(93)90162-r

Cited by 34 publications

(14 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Faecal cultures from goats were incubated at 27°C to recover infective third-stage larvae (L3) after 6-7 days [26]. L3 were exsheathed by exposure to 0.2% sodium hypochlorite (NaOCl) bubbled with air for 30 min at room temperature [27]. Exsheathed L3 (xL3) were separated from cuticular casts by migration through two 20 μm nylon meshes.…”

Section: Methodsmentioning

confidence: 99%

“…Exsheathed L3 (xL3) were separated from cuticular casts by migration through two 20 μm nylon meshes. xL3 were axenised in antibiotic solution (0.6 mg/ml penicillin, 1 mg/ml streptomycin, 40 μg/ml gentamycin and 10 μg/ml amphothericin B), then suspended in RPMI 1640 medium containing 20% (v/v) fetal bovine serum placed in culture flasks (175 cm 2 , vented cap, Corning) at a concentration of 1,000-2,000 larvae/ml and incubated at 40°C in 20% CO 2 for 7 days to produce early fourth-stage larvae (L4) [27]. The presence of a majority of individuals with L4 stage mouthparts, as described by Sommerville [28] and Mapes [29], was confirmed by microscopy.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Galectin Hco-gal-m from Haemonchus contortus modulates goat monocytes and T cell function in different patterns

Wang

Zhang

et al. 2014

Parasites Vectors

View full text Add to dashboard Cite

BackgroundMonocytes and T cells are two major subpopulations of peripheral blood mononuclear cells (PBMC) and play an essential role in the innate and adaptive immune systems. Different members of the galectin family show multiple and distinct regulatory effects on different cell types. Previous studies have demonstrated that the galectin from Haemonchus contortus (Hco-gal-m) performed immunomodulatory effects on goat PBMC, however, which subpopulation of PBMC is the primary target of Hco-gal-m and whether the immune modulations share the same mechanism remain unclear.MethodsIn this study, the developmental expression of Hco-gal-m was analyzed by RT-PCR and Western blot analysis. The distribution of Hco-gal-m in adult worm was detected by an immunohistochemical test. The binding activity of the recombinant Hco-gal-m (rHco-gal-m) on goat monocytes and T cells were assessed by flow cytometry. The immunomodulatory effects of Hco-gal-m on cytokine secretion, cell activation and apoptosis were observed by co-incubation of rHco-gal-m with goat monocytes and T cells.ResultsHco-gal-m was expressed in L4 as well as adult worms and predominantly localized at the internal surface of the worm guts. rHco-gal-m could bind to both monocytes and T cells. The engagement of rHco-gal-m decreased the production of IL-6, IL-10 and TNF-α in T cells, however, it significantly increased the secretion of IL-10 in monocytes. After rHco-gal-m exposure, the expression of MHC-II on monocytes and that of CD25 on T cells were restricted. Consequently, T cell proliferations were potently inhibited by rHco-gal-m. In addition, rHco-gal-m induced apoptosis in T cells, but not significantly in monocytes.ConclusionsOur results indicated that rHco-gal-m modulated goat monocytes and T cell function in different patterns.Electronic supplementary materialThe online version of this article (doi:10.1186/1756-3305-7-342) contains supplementary material, which is available to authorized users.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Galectin Hco-gal-m from Haemonchus contortus modulates goat monocytes and T cell function in different patterns

Wang

Zhang

et al. 2014

Parasites Vectors

View full text Add to dashboard Cite

show abstract

“…For infection, larvae were counted by a dilution technique. Exsheathment of L3 worms (xL3s) was carried out with NaOCl using the methods described by Rothwell and Sangster (1993). Haemonchus contortus diapause ZJ strains were obtained according to Blitz and Gibbs (1971).…”

Section: Propagation Of H Contortus and C Elegansmentioning

confidence: 99%

Hc-fau, a novel gene regulating diapause in the nematode parasite Haemonchus contortus

Yan

Guo

Zhou

et al. 2014

International Journal for Parasitology

View full text Add to dashboard Cite

“…The L1 were sterilized in RPMI 1640 medium (Biological Industries) containing 25 mM HEPES (Solarbio, Beijing, China), 1% fetal bovine serum and an antibiotic-antimycotic solution (10 μg/ml amphotericin B, 1 mg/ml streptomycin, 0.04 mg/ml gentamycin and 0.1 mg/ml carbenicillin) in a six-well plate (Corning, New York, USA) with constantly gentle shaking for 3 h at 28 °C. Unsheathment of L3s were performed with sodium hypochlorite as described previously [28]. The exsheathed L3s were washed in PBS supplemented with the antibiotic-antimycotic solution mentioned above in a 6-well plate and were kept in gentle shaking for 6 h at 37 °C.…”

Section: Heme Assaymentioning

confidence: 99%

Hc-hrg-2, a glutathione transferase gene, regulates heme homeostasis in the blood-feeding parasitic nematode Haemonchus contortus

Zhou

Zhao

et al. 2020

Parasites Vectors

View full text Add to dashboard Cite

Background: Haemonchus contortus, a blood-feeding parasite, is constantly surrounded by large quantities of heme released from the catabolism of host red blood cells. To cope with the toxicity of free heme, H. contortus needs to uptake and detoxify the heme, a process believed to be paramount for parasite survival. Methods: A heme-responsive gene Hc-hrg-2 was identified which is the homologue of Ce-hrg-2. The transcriptional levels in all developmental stages and heme-responsive ability of Hc-hrg-2 were analyzed by qRT-PCR. Immunofluorescence analysis and cell transfections were performed to analyze the expression pattern of Hc-HGR-2. Statistical analyses were performed with GraghPad Prism 6.0 using Student's t-test. Results: To investigate the heme homeostasis of H. contortus, we first identified a heme-responsive gene Hc-hrg-2, a homolog of Ce-hrg-2 that is involved in heme transport in the hypodermis of Caenorhabditis elegans. Using qRT-PCR, we showed that Hc-hrg-2 mRNA was expressed throughout all life-cycle stages of H. contortus with the highest level in the third-stage larvae (L3s). Notably, transcription of Hc-hrg-2 in the exsheathed L3s was significantly upregulated in the presence of high concentration of heme. We found that Hc-HRG-2 protein was mainly located in the hypodermal tissues of adult H. contortus in vivo and the endoplasmic reticulum in the transfected mammalian cells. Our in vitro assay demonstrated that Hc-HRG-2 is a heme-binding protein with glutathione S-transferase activity and heme had a significant effect on its enzymatic activity when a model substrate 1-chloro-2, 4-dinitrobenzene (CDNB) was used. Conclusions: Hc-hrg-2 is a heme-responsive gene and engaged in heme homeostasis regulation in hypodermal tissues during the free-living stages of H. contortus.

show abstract

An in vitro assay utilising parasitic larval Haemonchus contortus to detect resistance to closantel and other anthelmintics

Cited by 34 publications

References 18 publications

Galectin Hco-gal-m from Haemonchus contortus modulates goat monocytes and T cell function in different patterns

Galectin Hco-gal-m from Haemonchus contortus modulates goat monocytes and T cell function in different patterns

Hc-fau, a novel gene regulating diapause in the nematode parasite Haemonchus contortus

Hc-hrg-2, a glutathione transferase gene, regulates heme homeostasis in the blood-feeding parasitic nematode Haemonchus contortus

Contact Info

Product

Resources

About