Biomedical subjects

M J Doenhoff

Publications and source records attributed to M J Doenhoff.

At least 19 recordsLinked to original sources

PubMed2006

IPSE/alpha-1: a major immunogenic component secreted from Schistosoma mansoni eggs.

During infection with Schistosoma mansoni the egg stage of this parasite modulates the initial T helper (Th1) response into a Th2 response. This suggests that schistosome eggs contain factors responsible for that effect. We have recently described a glycoprotein (IPSE) from S. mansoni eggs that has a potent IL-4-inducing effect on human basophils. Here we demonstrate that IPSE is identical to a previously described molecule, the S. mansoni egg antigen alpha-1. We furthermore show that the expression of IPSE/alpha-1 at the level of both mRNA and protein is restricted to the egg stage. IPSE/alpha-1 is produced in and released from the subshell area of the egg and comes into close contact with inflammatory cells recruited to the vicinity of the egg surface. In line with this IPSE/alpha-1 is one of three major S. mansoni egg glycoproteins that induce pronounced antibody responses. Its IL-4-inducing capacity, moreover, suggests that IPSE/alpha-1 plays a role in initiating the Th2 response induced by patent S. mansoni infections.

Animals↗

PubMed2005

The role of acidic organelles in the development of schistosomula of Schistosoma mansoni and their response to signalling molecules.

The cercariae of Schistosoma mansoni become transformed into schistosomula during host skin penetration. We have found that large acidophilic compartments are detected in schistosomula but not in cercariae or in any other stages of the parasite by use of the fluorescent dye LysoTracker, a dye specific for mammalian lysosomes. Some of these large acidic compartments incorporated monodansylcadaverine, a specific dye for autophagosomes. We have used potent inhibitors (wortmannin and 3-methyladenine) and a potent inducer (starvation) of autophagy to show that the pathway to the formation of the acidic compartments requires specific molecular signals from the environment and from the genome. Certain doses of ultraviolet light inhibited significantly the formation of the acidic compartments, which may indicate disruption of the lysosome/autophagosome pathway. We have also defined two proteins that are commonly associated with lysosomes and autophagosomes in mammalian cells, the microtubule-associated membrane protein (MAP-LC3) and lysosome-associated membrane protein (LAMP-1), in extracts of schistosomula. We suggest that the autophagy pathway could be developed in transformed schistosomula.

Adenine↗

PubMed2005

Schistosoma japonicum reveals distinct reactivity with antisera directed to proteases mediating host infection and invasion by cercariae of S. mansoni or S. haematobium.

Serine proteases released from the acetabular glands of cercariae, also known as cercarial elastases, are key enzymes in the penetration process of schistosomes through the skin of the final host. Antisera against these enzymes secreted from Schistosoma mansoni or S. haematobium reveal differences in the patterns of elastase expression among schistosome species and among different developmental stages of the larvae. Immunolocalization studies showed that antisera raised against the enzyme s28 protease react with S. mansoni, S. haematobium and also S. japonicum, in developing as well as mature cercariae and in both pre- and post-acetabular glands. Antisera against the enzyme SmCE detect the respective antigen solely in the pre-acetabular glands. Remarkably, the SmCE-1a isoform is detectable with DNA-vaccinated mouse sera in S. mansoni and S. haematobium only, but is apparently absent from the acetabular glands of S. japonicum. These differences in immunoreactivity of cercarial enzymes may be related to the distinct infection process of S. japonicum.

Animals↗

PubMed2004

Echinococcus multilocularis metacestode extract triggers human basophils to release interleukin-4.

Infections with parasitic helminths are associated with a T helper 2 (Th2) immune response and IgE production. The underlying mechanism, however, is only partially understood. Recently we have isolated a protein from extracts of Schistosoma mansoni eggs that triggers human basophils from non-sensitized donors to release interleukin-4 (IL-4), the key cytokine of a Th2 response. We called this protein IPSE (for IL-4-inducing principle from Schistosoma mansoni eggs). Supposing that IPSE-like IL-4-inducing activities might be a general principle shared among different helminth species, we investigated extracts from the cestode E. multilocularis for its effect on human basophils. Our results showed that extracts from metacestodes of E. multilocularis cause basophil degranulation, as well as the secretion of histamine, IL-4 and IL-13, in a dose-dependent manner. IgE stripping and resensitization of basophils indicated that the mechanism of IL-4 induction requires the presence of IgE on the cells. Since analogous properties have been demonstrated earlier for IPSE, we think that S. mansoni and E. multilocularis may induce a Th2 response in their hosts via a related mechanism, namely, by the induction of IL-4 release from basophils.

Animals↗

PubMed2004

Serological speciation of human schistosome infections by ELISA with a panel of three antigens.

AIMS: To find out whether serology can reliably speciate human schistosomiasis using a simple enzyme linked immunosorbent assay (ELISA) technique. METHODS: Stored sera from 66 patients with microscopically confirmed schistosomiasis were subjected to ELISA using a panel of three antigens, namely: unfractionated Schistosoma mansoni soluble egg antigen (SEA); CEF6, a cationic fraction of SEA; and crude S margrebowiei egg antigen, prepared from an animal schistosome closely related to S haematobium. RESULTS: The optical densities (ODs) obtained using CEF6 as antigen were significantly higher in sera from S mansoni infected patients than in sera from S haematobium infected patients (median OD, 0.810 v 0.595). Using S margrebowiei egg antigen, the optical densities were significantly higher in S haematobium sera than in S mansoni sera (median OD, 0.794 v 0.544). There was no significant difference in optical densities between S mansoni and S haematobium sera using SEA (median OD, 0.725 v 0.737). The ratio of ODs (CEF6 to S margrebowiei egg antigen) was calculated: a ratio of >1 indicated S mansoni infection (sensitivity, 88%) and a ratio of <1 indicated S haematobium infection (sensitivity, 84%). The odds ratio for S haematobium having an OD ratio of <1 was 36.8 (95% confidence interval, 7.0 to 194). CONCLUSIONS: The identity of the infecting species of schistosome can be determined using the panel of antigens described. SEA should be used to screen serum samples, and the CEF6 : S margrebowiei egg antigen ELISA optical density ratio can be used where serological speciation is required.

Animals↗

PubMed2003

Identification of a fibrinolytic enzyme in Schistosoma mansoni eggs and modulated blood fibrinogen metabolism in S. mansoni-infected mice.

Aqueous extracts of Schistosoma mansoni eggs have been shown to have fibrinolytic activity inhibitable by a serine protease inhibitor. Fibrinolytic activity was not present in extracts of either adult worms or cercariae. A 27 kDa enzyme that was proteolytically active on fibrinogen in zymography and that degraded fibrinogen in a pattern similar to that of plasmin, is presumed to be responsible for the schistosome egg fibrinolytic activity. Anti-human fibrinogen antisera were shown to have antibodies that cross-reacted with mouse fibrinogen in Western immunoblots. Electroblotted sera from S. mansoni-infected and control uninfected mice displayed different antigenic profiles when probed with the cross-reactive anti-human fibrinogen antibodies, suggesting an alteration in mouse host fibrinogen metabolism as a result of the parasitic infection. We discuss the possibility that modulation of fibrinogen metabolism is a factor in a recently discovered anti-atherogenic effect exerted by schistosomes.

Animals↗

PubMed2003

Immune-dependent thrombocytopaenia in mice infected with Schistosoma mansoni.

As has been shown previously, immunologically intact mice with patent Schistosoma mansoni infections had a significantly lower mean platelet number than intact uninfected mice (P<0.0001). However, platelet numbers in T-cell deprived mice with patent infections were not significantly different from those in uninfected T-cell deprived mice. Also, platelet counts in both the infected and uninfected T-cell deprived groups were not significantly different from those in intact uninfected mice. The S. mansoni-induced thrombocytopaenia in mice is thus seemingly immune dependent. Immunologically intact mice with chronic 12-week-old S. mansoni infections had IgG antibodies that were reactive in an ELISA-type assay with whole fixed platelets of both mouse and human origin. In Western immunoblots the IgG antibodies from chronically-infected mice reacted in particular against mouse and human platelet antigens of 90, 37 and 30 kDa. Antisera raised from 2 rabbits, immunized respectively with mouse and human platelet antigens, cross-reacted with antigens of the larval, adult worm and egg stages of S. mansoni. These results support the hypothesis that an anti-platelet antibody response may be the cause of the thrombocytopaenia observed in mice with patent schistosome infections.

Animals↗

PubMed2003

The properties of acidic compartments in developing schistosomula of Schistosoma mansoni.

A variety of fluorescent probes have been used to study the acidic compartments in cercariae and schistosomula of Schistosoma mansoni. Freshly transformed schistosomula treated with the LysoTracker Red dye specific for lysosomes showed large acid-containing compartments (0.5-10 microm in size). The uptake of the dye is an energy-dependent process that depends on the metabolic activity of schistosomula. The compartments were quantified individually with respect to area, quantity of fluorescence and the total number/schistosomulum. Under normal conditions these compartments were not found in untreated cercariae, but appeared in cercariae slightly damaged by poly-L-lysine. The formation of these compartments seemed to be related to the development of cercariae into schistosomula as the number of compartments and uptake of fluorescence increased with time after transformation. Also, the method of transformation as well as the in vitro incubation of the parasite affected the percentage area of compartments/schistosomulum. Acid phosphatase enzyme activity was assessed using an endogenous phosphatase probe. Living and fixed schistosomula displayed the presence of enzyme activity in compartments of the same size and distribution as the acid-rich compartments. This was confirmed by histochemical staining showing deposition of enzyme-generated lead at the sites of phosphatase activity. We suggest that the development of acidic compartments is important during the transformation process or as a consequence of damage.

Acid Phosphatase↗

PubMed2003

Phylogeography of Biomphalaria glabrata and B. pfeifferi, important intermediate hosts of Schistosoma mansoni in the New and Old World tropics.

The historical phylogeography of the two most important intermediate host species of the human blood fluke Schistosoma mansoni, B. glabrata in the New World, and B. pfeifferi in the Old World, was investigated using partial 16S and ND1 sequences from the mitochondrial genome. Nuclear sequences of an actin intron and internal transcribed spacer (ITS)-1 were also obtained, but they were uninformative for the relationships among populations. Phylogenetic analyses based on mtDNA revealed six well-differentiated clades within B. glabrata: the Greater Antilles, Venezuela and the Lesser Antilles, and four geographically overlapping Brazilian clades. Application of a Biomphalaria-specific mutation rate gives an estimate of the early Pleistocene for their divergence. The Brazilian clades were inferred to be the result of fragmentation, due possibly to climate oscillations, with subsequent range expansion producing the overlapping ranges. Within the Venezuela and Lesser Antilles clade, lineages from each of these areas were estimated to have separated approximately 740 000 years ago. Compared to B. glabrata, mitochondrial sequences of B. pfeifferi are about 4x lower in diversity, reflecting a much younger age for the species, with the most recent common ancestor of all haplotypes estimated to have existed 880 000 years ago. The oldest B. pfeifferi haplotypes occurred in southern Africa, suggesting it may have been a refugium during dry periods. A recent range expansion was inferred for eastern Africa less than 100 000 years ago. Several putative species and subspecies, B. arabica, B. gaudi, B. rhodesiensis and B. stanleyi, are shown to be undifferentiated from other B. pfeifferi populations.

Africa↗

PubMed2003

The detection of antibodies against Schistosoma mansoni soluble egg antigens (SEA) and CEF6 in ELISA, before and after chemotherapy.

Circulating IgG antibody reactivity and excreted egg counts were investigated in 489 Kenyans given chemotherapy for schistosomiasis mansoni. Antibody reactivity was measured in ELISA, using either unfractionated aqueous soluble constituents of Schistosoma mansoni eggs (SEA) or CEF6 (a soluble fraction of S. mansoni eggs containing two cationic antigens) as the antigen source. Antibody reactivity for each antigen source was strongly associated with egg counts, both pre- and post-treatment. Approximately 6 months after chemotherapy, egg counts were zero in 84% of the subjects. The mean optical densities (OD) measured in the post-treatment ELISA were 60% (CEF6) or 45% (SEA) lower than the pre-treatment values, the reduction in the OD with CEF6 as antigen source being significantly greater than that observed with SEA (P <0.001). The usefulness of an assay for antibody reactivity in monitoring the effects of the treatment of schistosomiasis is discussed.

Adolescent↗

PubMed2002

An anti-atherogenic effect of Schistosoma mansoni infections in mice associated with a parasite-induced lowering of blood total cholesterol.

In affluent societies the prevalences of so-called 'Western' diseases such as atherosclerosis, allergies and autoimmune disorders appear to have increased, while many diseases caused by communicable infections are now relatively less common. To test whether there may be a causal relationship we examined the effects of Schistosoma mansoni infections in mice that develop cardiovascular pathology as a result of a genetic deficiency in apolipoprotein E (apoE-/-). The development of atherosclerotic lesions in the aortic arch and brachiocephalic artery of the apoE-/- mice was reduced by approximately 50% in mice with the parasitic infection, when comparison was made with uninfected control mice fed the same diet. Observations on S. mansoni-infected conventional laboratory mice indicate that patent schistosome infections could be counteracting the effects of an atherogenic diet by modulating host lipid metabolism and inducing a reduction in blood total cholesterol concentrations.

Animals↗

PubMed2002

Adult worm tegumental damage and egg-granulomas in praziquantel-resistant and -susceptible Schistosoma mansoni treated in vivo.

The effect of treatment with praziquantel (PZQ) on the tegument of adult Schistosoma mansoni worms and on liver egg-granulomas has been examined in mice infected with PZQ-resistant and -susceptible parasite isolates. Two PZQ-resistant S. mansoni isolates, one selected by passage in the laboratory under drug pressure and one from Senegal established from eggs excreted by an uncured patient, were compared with PZQ-susceptible control isolates. Scanning electron microscopic observations on the tegument of Schistosoma adult worms treated in vivo with PZQ showed that more severe damage was inflicted by PZQ on susceptible worms than on drug-resistant worms. Observations on the pathology of Schistosoma egg-granulomas in the liver of infected mice after treatment with PZQ indicated that eggs from susceptible control isolates were more sensitive to PZQ than those from drug-resistant isolates.

Animals↗

PubMed2001

Acidic vesicles of Schistosoma mansoni.

The fluorescent probe LysoTracker Red was used to examine for the presence of acidic vesicles in cercariae and schistosomula of Schistosoma mansoni. Acidic vesicles were widely distributed throughout the body of freshly transformed schistosomula and 24-h-old schistosomula but were absent from cercariae. The vesicles of freshly transformed schistosomula were undetectable after incubation with drugs that affect the functionality of acidic vesicles including monensin, ouabain, primaquine, and amiloride. In 24-h transformed schistosomula, the same effect was observed with monensin but not with ouabain, primaquine or amiloride. Praziquantel also affected the acidic vesicles of the schistosomula. We suggest that these acidic vesicles could be large lysosome-like organelles.

Animals↗

PubMed2001

In vitro responses of praziquantel-resistant and -susceptible Schistosoma mansoni to praziquantel.

The resistance status of five praziquantel-susceptible and five praziquantel-resistant isolates was confirmed by chemotherapy in CD(1) mice with 3 x 200mg/kg micronised praziquantel. Micronised praziquantel had higher efficacy than two other praziquantel formulations (prepared without milling). The five resistant isolates were less responsive to praziquantel than the five susceptible isolates (59-74% reduction in worm burden in resistant isolates compared with 92-100% in susceptible isolates). Observations were made on the in vitro responses of different stages of 10 isolates to praziquantel. There were different in vitro responses to praziquantel at the egg, miracidial, cercarial and adult stages of Schistosoma mansoni between praziquantel-resistant and praziquantel-susceptible isolates. There were differences in the response of resistant and susceptible isolates following exposure of freshly hatched miracidia to 10(-6)M praziquantel for 1 min and observing the percent change in shape. Using this test it should be possible to determine whether failed therapy in patients infected with S. mansoni is due to the presence of praziquantel-resistant worms. Similarly, by exposing freshly shed cercariae to 4 x 10(-7)M praziquantel and observing the percent of tail shedding over 80 min it should be possible to monitor for the presence of praziquantel-resistant worms in snails collected in the field.

Animals↗

PubMed2001

Infection induces antibodies against the cercarial secretions, but not against the cercarial elastases of Schistosoma mansoni, Schistosoma haematobium, Schistosoma japonicum and Trichobilharzia ocellata.

Cercarial secretions from different species of the parasite Schistosoma and from Trichobilharzia ocellata contain a proteolytic activity, cercarial elastase, which was demonstrated by a 30 kDa band in gelatin gels. Sera of patients infected with Schistosoma mansoni, Schistosoma haematobium or Schistosoma japonicum contain immunoglobulin G which react in ELISA with cercarial secretions from all schistosomes and cross-react among the different parasite species. In Western blots, however, infection sera from patients, as well as heavily infected mice or rabbits, did not react with a 30-kDa protein. Moreover, when sections from infected snails (Biomphalaria, Bulinus and Lymnaea) were analysed by immunofluorescence using the same infection sera, only the tegument of the developing cercariae was recognized, but not the acetabular glands. In contrast, when antisera against purified cercarial elastase from either S. mansoni or S. haematobium were tested with sections of infected Biomphalaria or Bulinus, fluorescence was strong in the preacetabular glands of the cercariae of either species, but undetectable with the tegument. Cross-reactivity of both antisera extended to T. ocellata-infected Lymnaea, but not to S. japonicum-infected Oncomelania. In conclusion, although immunization with purified cercarial elastase results in antibody production, the enzyme does not induce an apparent antibody response following natural infection.

Animals↗

PubMed2001

Biological characteristics of praziquantel-resistant and -susceptible isolates of Schistosoma mansoni.

If there is a change in the biological characteristics of schistosomes associated with the development of resistance to praziquantel, this could affect the transmission and pathology of the diseases they cause. To investigate this possibility, the host-parasite relationships of five praziquantel-resistant and five praziquantel-susceptible isolates of Schistosoma mansoni were compared in Biomphalaria glabrata snails and outbred CD(1) albino mice. Whether praziquantel-resistant or -susceptible, all the laboratory-selected isolates gave similar results in B. glabrata. However, the snails infected with any of three field-collected isolates from Senegal, each of which has been shown to be praziquantel-resistant, shed fewer cercariae and survived longer than the snails infected with the drug-susceptible or laboratory-selected, drug-resistant isolates. There were no differences between isolates in terms of their infectivity to mice. However, mice infected with any of four of the five drug-resistant isolates shed more eggs in their faeces than mice carrying the drug-susceptible parasites, and mice infected with any of the resistant isolates also had larger numbers of eggs in their tissues. Although granuloma sizes were generally similar for all isolates, the praziquantel-resistant isolates may be more pathogenic in mice than the susceptible ones because of their relatively high egg productions.

Animals↗

PubMed2000

Stage-associated expression of ceramide structures in glycosphingolipids from the human trematode parasite Schistosoma mansoni.

Glycosphingolipids of Schistosoma mansoni adults, cercariae and eggs comprise ceramide monohexosides (CMH) with glucose or galactose and ceramide dihexosides (CDH) with the schistosome-specific structure GalNAc(beta1-4)Glc(1-1)ceramide. Ceramide analysis revealed C18- and C20-phytosphingosines in egg CMH, C18-sphinganine as well as C18-, C19- and C20-phytosphingosines in cercarial CMH, and C18- and C20-phytosphingosines as well as C18-sphingosine and C18-sphinganine in adult CMH. For all three life cycle stages, the predominant fatty acid was C16h:0. As a characteristic feature, a range of saturated, unsaturated and hydroxylated long-chain fatty acids with 24-28 carbon atoms were additionally found in minor cercarial CMH species. The corresponding ceramides represented major constituents in cercarial CDH, while adult and egg CDH were dominated by ceramides with short fatty acid chains. The resultant ceramide patterns could be correlated with the differential expression of carbohydrate antigens on schistosomal glycolipids at various stages. A possible impact of ceramide structure on the biosynthesis of the carbohydrate moieties is discussed.

Animals↗

PubMed2000

A fucose-containing epitope is shared by keyhole limpet haemocyanin and Schistosoma mansoni glycosphingolipids.

The glycolipids of Schistosoma mansoni adult worms, cercariae and eggs are recognised by schistosome infection serum and the monoclonal antibody M2D3H. The haemocyanin of the keyhole limpet, Megathura crenulata, is known to be immunoreactive to schistosomal infection sera and is, therefore, under investigation for the diagnosis of and vaccination against schistosomiasis. By dot-blot, inhibition-ELISA and inhibition-HPTLC immunostaining we have demonstrated that the M2D3H epitope is shared by both S. mansoni glycolipids and keyhole limpet haemocyanin (KLH). Analogously to the established epitopic importance of fucose to the immunorecognition of S. mansoni glycolipids, we have similarly defined the significance of the fucose residue(s) for the immunoreactivity between KLH and schistosomal infection serum and the monoclonal antibody M2D3H. Fucose was specifically removed from KLH by partial hydrolysis, monitored by ultrafiltration and carbohydrate component analysis. On removal of the fucose residue(s) the serological and immunological reactivity of KLH was greatly diminished, which implied that the fucose-containing M2D3H antigenic determinant was common to both S. mansoni glycolipids and KLH.

Animals↗