Schistosoma mansoni excretory–secretory products stimulate a p38 signalling pathway in Biomphalaria glabrata embryonic cells

Humphries, Judith E.; Yoshino, Timothy P.

doi:10.1016/j.ijpara.2005.08.009

Cited by 34 publications

(27 citation statements)

References 53 publications

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“…Bge cells and B. glabrata hemocytes were isolated as previously described (Humphries and Yoshino, 2006). Total RNA from both cell sources was extracted with Trizol (Invitrogen Corporation, Carlsbad, CA), followed by precipitation with isopropanol according to the manufacturer's protocol and stored at −80°C until further use.…”

Section: Rna Extraction and Rapid 5' And 3' Amplification Of Cdna Endmentioning

confidence: 99%

“…The recovered plasma was filtered (0.45 µm), concentrated by centrifugation using first a 100 kDa cut-off spin-filter to remove the majority of hemoglobin, and then a 10 kDa cut-off filter (Millipore) to concentrate any BgGal. All protein samples were subjected to SDS-PAGE gels (12.5%), and assayed for the presence of galectin by Western blot analysis (Humphries and Yoshino, 2006).…”

Section: Collection and Extraction Of Proteins From Snail Hemocytes Amentioning

confidence: 99%

“…Protocols used for Western immunoblotting were similar to those previously described (Humphries and Yoshino, 2006). Briefly, snail proteins were separated on a 12.5% SDS-PAGE gel, followed by transfer to a nitrocellulose membrane (Hybond ECL; Amersham Pharmacia Biotech) by semi-dry electroblotting, then blocked overnight in Tris-buffered saline (sTBS; 20 mM Tris, 45 mM NaCl, pH 7.4) containing 5% dry milk, and reacted with anti-rBgGal antibodies (diluted 1:5000 for hemocytes; 1:2000 for plasma proteins; 1:2500 for Bge cell extracts) or rabbit pre-immune serum (1:5000) for 16 hr at 4°C.…”

Section: Western Blot Analysismentioning

confidence: 99%

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Molecular and functional characterization of a tandem-repeat galectin from the freshwater snail Biomphalaria glabrata, intermediate host of the human blood fluke Schistosoma mansoni

Yoshino

Dinguirard

Kunert

et al. 2008

Gene

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In the present study, a tandem-repeat type galectin was characterized from an embryonic cell line (Bge) and circulating hemocytes of the snail Biomphalaria glabrata, intermediate host of the human blood fluke Schistosoma mansoni. The predicted B. glabrata galectin (BgGal) protein of 32 kDa possessed 2 carbohydrate recognition domains, each displaying 6 of 8 conserved amino acids involved in galactoside-binding activity. A recombinant BgGal (rBgGal) demonstrated hemagglutinating activity against rabbit erythrocytes, which was specifically inhibited by galactosecontaining sugars (lacNAc/lac > galNAc/gal). Although native galectin was immunolocalized in the cytoplasm of Bge cells and the plasma membrane of a subset of snail hemocytes (60%), it was not detected in cell-free plasma by Western blot analysis. The findings that rBgGal selectively recognizes the schistosome-related sugar, lacNAc, and strongly binds to hemocytes and the tegument of S. mansoni sporocysts in a sugar-inhibitable fashion suggest that hemocyte-bound galectin may be serving as pattern recognition receptor for this, or other pathogens possessing appropriate sugar ligands. Based on molecular and functional features, BgGal represents an authentic galectin, the first to be fully characterized in the medically-important molluscan Class Gastropoda.

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Section: Rna Extraction and Rapid 5' And 3' Amplification Of Cdna Endmentioning

confidence: 99%

Section: Collection and Extraction Of Proteins From Snail Hemocytes Amentioning

confidence: 99%

Section: Western Blot Analysismentioning

confidence: 99%

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Molecular and functional characterization of a tandem-repeat galectin from the freshwater snail Biomphalaria glabrata, intermediate host of the human blood fluke Schistosoma mansoni

Yoshino

Dinguirard

Kunert

et al. 2008

Gene

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“…Excretory-secretory products (ESP) used in initial experiments were harvested from the 48-hr in vitro cultured S. mansoni sporocysts as described in Humphries and Yoshino [11].…”

Section: Schistosoma Mansoni Parasite Culture and Harvesting Of Excrementioning

confidence: 99%

Regulation of hydrogen peroxide release in circulating hemocytes of the planorbid snail Biomphalaria glabrata

Humphries

Yoshino

2008

Developmental & Comparative Immunology

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Biomphalaria spp. serve as obligate intermediate hosts for the human blood fluke Schistosoma mansoni. Following S. mansoni penetration of Biomphalaria glabrata, hemocytes of resistant snails migrate towards the parasite, encasing the larva in a multicellular capsule resulting in its destruction via a cytotoxic reaction. Recent studies have revealed the importance of hydrogen peroxide and nitric oxide (H 2 O 2 , NO) in parasite killing [1,2]. It is assumed H 2 O 2 and NO production is tightly regulated although the specific molecules involved remain largely unknown. Consequently, the potential role of cell signaling pathways in B. glabrata hemocyte H 2 O 2 production was investigated by evaluating the effects of specific inhibitors of selected signaling proteins. Results suggest that both ERK and p38 MAPKs are involved in the regulation of B. glabrata H 2 O 2 release in response to stimulation by PMA and galactose-conjugated BSA. However, the involvement of the signaling proteins PKC, PI 3 kinase and PLA 2 differs between PMA-and BSA-gal-induced H 2 O 2 production.

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“…Better understanding the fundamental mechanism of snail internal defense and associated immune signaling pathways will provide new insights that could lead to innovative control strategies for blocking the transmission of schistosomiasis within the snail host and, at the same time, broaden our knowledge of the evolution of innate immunity. Recently, considerable progress has been made in exploring the mechanisms of snail internal defense at the molecular level Vergote et al 2005;Goodall et al 2006;Humphries and Yoshino 2006;Raghavan and Knight 2006;Lockyer et al 2007), but still much remains unknown. This paper focuses on our recent studies of B. glabrata genes encoding peptidoglycan (PGN) recognition proteins (PGRPs; including long and short forms) and gram-negative bacteria binding protein (GNBP).…”

Section: Introductionmentioning

confidence: 99%

Characterization of immune genes from the schistosome host snail Biomphalaria glabrata that encode peptidoglycan recognition proteins and gram-negative bacteria binding protein

2007

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Peptidoglycan (PGN) recognition proteins (PGRPs) and gram-negative bacteria binding proteins (GNBPs) play an essential role in Toll/Imd signaling pathways in arthropods. The existence of homologous pathways involving PGRPs and GNBPs in other major invertebrate phyla such as the Mollusca remains unclear. In this paper, we report four full-length PGRP cDNAs and one fulllength GNBP cDNA cloned from the snail Biomphalaria glabrata, the intermediate host of the human blood fluke Schistosoma mansoni, designated as BgPGRPs and BgGNBP, respectively. Three transcripts are generated from a long form PGRP gene (BgPGRP-LA) by alternative splicing and one from a short form PGRP gene (BgPGRP-SA). BgGNBP encodes a putative secreted protein. Northern blots demonstrated that expression of BgPGRP-SA and BgGNBP was down-regulated in B. glabrata at 6 h after exposure to three types of microbes. No significant changes in expression were observed in snails at 2 days post-exposure (dpe) to the trematodes Echinostoma paraensei or S. mansoni. However, up-regulation of BgPGRP-SA in M line snails at later time points of infection with E. paraensei (i.e., 12 and 17 dpe) was observed. Our study revealed that exposure to either microbes or trematodes did not alter the expression levels of BgPGRP-LAs, which were consistently low. This study provides new insights into the potential pathogen recognition capabilities of molluscs, indicates that further studies of the Toll/Imd pathways in this phylum are in order, and provides additional ways to judge the importance of this pathway in the evolution of internal defense across the animal phyla.

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Schistosoma mansoni excretory–secretory products stimulate a p38 signalling pathway in Biomphalaria glabrata embryonic cells

Cited by 34 publications

References 53 publications

Molecular and functional characterization of a tandem-repeat galectin from the freshwater snail Biomphalaria glabrata, intermediate host of the human blood fluke Schistosoma mansoni

Molecular and functional characterization of a tandem-repeat galectin from the freshwater snail Biomphalaria glabrata, intermediate host of the human blood fluke Schistosoma mansoni

Regulation of hydrogen peroxide release in circulating hemocytes of the planorbid snail Biomphalaria glabrata

Characterization of immune genes from the schistosome host snail Biomphalaria glabrata that encode peptidoglycan recognition proteins and gram-negative bacteria binding protein

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