Lung Schistosomula Research Articles

Schistosomiasis mansoni: A new therapeutic target for ubiquinol, a natural inhibitor of neutral magnesium-dependent sphingomyelinase in murine model.

Constituting the host-parasite interface and playing a censorious role in host immune response modulation and parasite survival, tegument represents a crucial target for many antischistosomal drugs. Sphingomyelin forms a stable outer leaflet of tegumental membrane-lipid bilayer. Neutral magnesium‏-dependent sphingomyelinase (Mg2+-nSMase) is a key enzyme in sphingomyelin breakdown was identified in schistosomes. We investigated the in vivo efficacy of ubiquinol, a natural inhibitor of Mg2+-nSMase, in free and niosomes-encapsulated forms, through five-day and 15-day regimens on the early and late Schistosoma mansoni parasitic stages, respectively, compared to PZQ. Oral administration of 300 mg/kg/day ubiquinol-encapsulated niosomes (U-N) showed significant deterioration of the parasitic growth and development in the term of reduction of lung schistosomula burden (39.12%), adult worm burden (50.81%), hepatic and intestinal tissue-egg counts (80.89% and 75.54%, respectively). PZQ and free ubiquinol regimens reported reductions in lung schistosomula counts (45.36% and 22.90%, respectively) and total worm burdens of 86.28% and 24.58%, respectively. U-N therapy revealed worms de-pairing and remarkable diminution in female worms’ perimeters and fecundity. Scanning electron microscope revealed disruption of tegumental ridges with excessive longitudinal corrugation. Transmission electron microscope showed testicular and ovarian parenchymal degeneration, signs of immaturity and cell apoptosis. Indirect immunofluorescence assay approved parasite's tegumental changes. Remarkable reduction of granulomas size with amelioration of hepatic pathology and fibrosis were assumed to be attributed to the anti-inflammatory and anti-oxidant properties of ubiquinol. These findings with the drug safety profile suggest that U-N could be a promising candidate for a new antischistosomal drug development.

Trials of vaccination by lung schistosomula and Biomphalaria alexandrina vaccines against experimental Schistosoma mansoni

Background Schistosomulum stage is believed to be the target of protective immunity. Over the past two decades, several investigators have demonstrated the antigenic communion between Schistosoma mansoni and Biomphalaria alexandrina. Objective The present study aimed to evaluate the effect of combining S. mansoni schistosomal lung antigen preparations and B. alexandrina antigen preparations for use as antischistosomal vaccination in murine models, and to compare their efficacy with and without the use of complete Freund’s adjuvant (CFA). Materials and methods Seventy laboratory-bred Swiss albino male mice were used in this study. They were classified into seven groups (10 mice each). Each mouse was sensitized with an initial subcutaneous injection of the extracted antigens. After 2 weeks, a second subcutaneous injection of the same antigen dose was given. Two weeks after the last dose of vaccination, all mice groups were infected with 100 S. mansoni cercariae subcutaneously. Mice were sacrificed by rapid decapitation 7 weeks post-infection for assessment of the inoculated antigens by parasitological (stool egg count, worm burden, tissue egg load, and oogram pattern) and histopathological (hepatic sections stained with hematoxylin and eosin for detection of granuloma number and diameter) studies. Results The data showed that vaccination with combined antigens (S. mansoni schistosomal lung antigen prepations + B. alexandrina antigen preparations with CFA) had the best protective effect. Conclusion The single antigen vaccination did not protect against infection by antigenically complexed S. mansoni. The cocktail vaccine apparently induced an agreeable immune response against many of the antigenic components. This new cocktail represents a promising approach toward the future development of vaccine strategy.

Up-regulation of SUMO E3 ligases during lung schistosomula and adult worm stages

Small ubiquitin-like modifier (SUMO) conjugation of proteins occurs through a concert action of enzymes using a similar ubiquitination mechanism. After a C-terminal peptide is cleaved from the SUMO precursor by a protease to reveal a di-glycine motif, SUMO is activated by an E1 enzyme (Aos1/Uba2) and conjugated to target proteins by the sole E2 enzyme (Ubc9) guided to the appropriate substrates by the SUMO E3 ligase. Previous reports from our group showed that Schistosoma mansoni has two paralogs of SUMO: one E2 conjugation Ubc9 and two SUMO-specific proteases (SENPs). The differential gene expression profile observed for SUMO pathway genes throughout the S. mansoni life cycle attests for the distinct patterns of SUMO conjugates observed during parasite development particularly during the cercariae to schistosomula transition. To continue this investigation, we here analysed the repertoire of SUMO E3 ligases and their expression profiles during cercariae/schistosomula transition. In silico analysis through S. mansoni databases showed two conserved SUMO E3 ligases: protein inhibitor of activated STAT (PIAS) and Ran-binding protein 2 (RanBP2). Furthermore, expression levels of the SUMO E3 ligases were measured by qRT-PCR using total RNA from cercariae, adult worms and mechanically transformed schistosomula. Our data showed an up-regulation of expression in lung schistosomula and adult worm stages. In conclusion, the differential expression of SmPIAS and SmRanBP2 during schistosomula development was similar to the expression levels of all genes related to SUMO conjugation, thereby suggesting that the control of protein activity, localisation or stability during cercariae to schistosomula transition is SUMO-dependent.

β-Lapachone: A naphthoquinone with promising antischistosomal properties in mice

The activity of β-lapachone (3,4-dihydro-2,2-dimethyl-2H-naphthol[1,2-b]pyran-5,6-dione, β-lap) against different stages of Schistosoma mansoni was investigated in mice. Mice infected with 50 cercariae (BH strain) were intraperitoneally treated at a dose of 50mg/kg for 5 consecutive days, starting on the 1st, 14th, 28th and 45th days after infection, to evaluate the effect of β-lap on skin schistosomula, lung schistosomula, young worms (before oviposition) and adult worms (after oviposition), respectively. All animals were euthanized 60 days after infection. β-Lap significantly reduced (p<0.001) the number of worms in 29.78%, 37.2%, 24.2% and 40.22% when administered during the phases of skin schistosomula, lung schistosomula, young worms and adult worms, respectively. Significant reduction was also achieved in terms of female burden. In all groups, there was significant reduction in the number of eggs and granulomas in the hepatic tissue. When the intervention was performed during the phase of adult worms, β-lap reduced the size of hepatic granulomas and changed the oogram pattern, lowering the percentage of immature eggs and increasing the percentage of mature and dead eggs. Our data indicate that β-lap has moderate antischistosomal properties. Its molecule may also be used as a prototype for synthesis of new naphthoquinone derivatives with potential schistosomicidal properties. Further studies with different formulations containing β-lap are needed to clearly establish the best dose and route of administration and its mechanism of action against schistosomes.

Identification of Th1 epitopes within molecules from the lung-stage schistosomulum of Schistosoma japonicum by combining prediction analysis of the transcriptome with experimental validation

The lung-stage schistosomulum has been regarded as the main target of protective immunity induced by radiation-attenuated vaccines (RAV) in the mouse model of schistosomiasis, and immune mechanisms mediated by the CD4+ Th1 response play a major role in the RAV model. To identify Th1 epitopes rapidly within molecules from the lung schistosomulum of Schistosoma japonicum, in the present study we analyzed transcriptome data from normal and radiation-attenuated lung schistosomula of S. japonicum and Schistosoma mansoni. We selected six genes with high levels of expression of their transcripts as sample sequences from the lung schistosomula. From these six sequences, by using different algorithms, we predicted six promiscuous Th cell epitopes that are capable of binding to both murine and human MHC class II molecules. To validate our in silico prediction experimentally, first, the gene expressions of the six sequences in day 3 lung-stage schistosomula were assessed using reverse-transcription PCR (polymerase chain reaction) analysis. The result showed that all six sequences predicted can be expressed in normal day 3 schistosomula. Second, we measured the direct binding of the four peptides predicted above to APCs (Antigen Presenting Cells) from the BALB/c mouse strain using a fluorometric method, and found that the four peptides could bind to both I-Ad and I-Ed molecules of the mice. Finally, the proliferation and profiles of cytokine production by spleen lymphocytes from the BALB/c mice immunized with the six predicted peptides were detected in vitro using modified MTT (Methyl Thiazolyl Tetrazolium), and flow cytometry methods, respectively. The results showed that three of the six predicted peptides could induce a recall CD4+ Th1 response in vitro. These results demonstrate that potential Th1-type epitopes can be identified rapidly by a combination of in silico analysis of transcriptomes of lung-stage schistosomula with experimental validation.

The saga of schistosome migration and attrition

Schistosomes infect the mammalian host by direct penetration of the skin and must then undergo a protracted migration to the site of parasitization, for Schistosoma mansoni the hepatic portal vasculature. This article reviews the work published roughly between 1976 and 1986 that clarified our understanding of the process in the laboratory mouse. A combination of histopathology, larval injection experiments and autoradiographic tracking revealed that migration involved one to several circuits of the pulmonary-systemic vasculature before chance delivery in cardiac output to splanchnic arteries that lead indirectly to the portal tract. The kinetics of migration through different capillary beds was established, with the lungs of naïve mice not the skin proving the greatest obstacle; a proportion of schistosomula entered the alveoli from where they did not recover. The 'immunity' displayed by mice with a chronic infection was shown to be an artefact of a 'leaky' hepatic portal system, generated as a result of egg-induced hepatic pathology. The blockade of pulmonary migration was exacerbated in mice vaccinated with irradiated cercariae by immune-mediated inflammatory foci that developed around lung schistosomula thus decreasing the proportion that matured, but parasite elimination was a prolonged process, not an acute cytolytic 'hit.'

Developmental gene expression profiles of the human pathogen Schistosoma japonicum

BackgroundThe schistosome blood flukes are complex trematodes and cause a chronic parasitic disease of significant public health importance worldwide, schistosomiasis. Their life cycle is characterised by distinct parasitic and free-living phases involving mammalian and snail hosts and freshwater. Microarray analysis was used to profile developmental gene expression in the Asian species, Schistosoma japonicum. Total RNAs were isolated from the three distinct environmental phases of the lifecycle – aquatic/snail (eggs, miracidia, sporocysts, cercariae), juvenile (lung schistosomula and paired but pre-egg laying adults) and adult (paired, mature males and egg-producing females, both examined separately). Advanced analyses including ANOVA, principal component analysis, and hierarchal clustering provided a global synopsis of gene expression relationships among the different developmental stages of the schistosome parasite.ResultsGene expression profiles were linked to the major environmental settings through which the developmental stages of the fluke have to adapt during the course of its life cycle. Gene ontologies of the differentially expressed genes revealed a wide range of functions and processes. In addition, stage-specific, differentially expressed genes were identified that were involved in numerous biological pathways and functions including calcium signalling, sphingolipid metabolism and parasite defence.ConclusionThe findings provide a comprehensive database of gene expression in an important human pathogen, including transcriptional changes in genes involved in evasion of the host immune response, nutrient acquisition, energy production, calcium signalling, sphingolipid metabolism, egg production and tegumental function during development. This resource should help facilitate the identification and prioritization of new anti-schistosome drug and vaccine targets for the control of schistosomiasis.

Schistosoma mansoni ex vivo lung-stage larvae excretory–secretory antigens as vaccine candidates against schistosomiasis

Schistosoma mansoni lung-stage larvae are known to be the major target of innate and acquired immunity to schistosomiasis. Lung schistosomula cytosolic or surface membrane antigens are hidden, entirely inaccessible to the host immune system, and hence are not particularly important as vaccine candidates. Conversely, excretory–secretory (E–S) products released from intact, viable, elongated, and contractile schistosomula are ideal potential vaccines, as such molecules can readily play a central role in the induction of local primary and memory immune response effectors that would directly target, surround, and pursue the larvae while negotiating the lung capillaries. Therefore, 6-day-old ex vivo larvae were isolated from mouse or hamster lung cells and used for generation of E–S products, which were shown to elicit strong immune responses and significant ( P < 0.05) protection against challenge infection in BALB/c mice. Proteomic analysis of E–S molecules following 10× concentration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis identified peptides related to innumerable host and about 15 S. mansoni-specific proteins. Selected S. mansoni-specific E–S peptides prepared in a multiple antigen peptide (MAP) or recombinant form were shown to stimulate considerable specific antibody response and peripheral blood mononuclear cell expression of mRNA for several cytokines in immunized C57BL/6 and BALB/c mice. However, highly significant ( P < 0.05 to <0.005) reduction in challenge infection worm burden and egg load was recorded only when the immunization conditions in test mice provided the S. mansoni antigen-specific T helper (Th) type response milieu favorable for each immunogen. That was polarized Th1 for S. mansoni aldolase and thioredoxin peroxidase 1 MAPs, polarized Th2 for recombinant 14-3-3-like protein, mixed Th1/Th17 for calpain MAP, and mixed Th1/Th2 for recombinant p18 protein. The findings together indicated that the immune responses issue is as critical as the nature and source of the antigen for the development of vaccine against schistosomiasis.

Patterns of gene expression in schistosomes: localization by whole mountin situhybridization

As a consequence of comprehensive transcriptome analysis followed by sequencing and draft assembly of the genome, the emphasis of schistosome research is shifting from the identification of genes to the characterization of their functions and interactions. Developmental biologists have long used whole mount in situ hybridization (WISH) to determine gene expression patterns, as a vital tool for formulating and testing hypotheses about function. This paper describes the application of WISH to the study of gene expression in larval and adult schistosomes. Fixed worms were permeablized by proteinase K treatment for hybridization with digoxygenin-labelled RNA probes, with binding being detected by alkaline phosphatase-coupled anti-digoxygenin antibodies, and BM Purple substrate. Discrete staining patterns for the transcripts of the molecules Sm29, cathepsin L, antigen 10.3 and chorion were observed in the tegument cell bodies, gut epithelium, oesophageal gland and vitelline lobules, respectively, of adult worms. Transcripts of the molecules SGTP4, GP18-22 and cathepsin L were localized to tegument cell bodies and embryonic gut, respectively, of lung schistosomula. We also showed that Fast Red TR fluorescent substrate can refine the pattern of localization permitting use of confocal microscopy. We believe that method of WISH will find broad application, in synergy with other emerging post-genomic techniques, such as RNA interference, to studies focused at increasing our molecular understanding of schistosomes.

Update on paramyosin in parasitic worms

Paramyosin was first identified as a structural component of invertebrate muscle. Analysis of crude, native, adult schistosome worm preparations identified a highly immunogenic protein which was later identified as paramyosin. Early vaccination/challenge studies with native paramyosin produced encouraging levels of protective efficacy against schistosomes, which led to the question as to how a sub-tegumental (muscular) protein could provide a target for vaccine-mediated immunological attack. Immunolocalisation studies of schistosomes confirmed the presence of paramyosin within the post-acetabular glands of cercariae and on the tegumental surface of lung schistosomula. Here we present an update on the more recent research on paramyosin in parasitic worms that has focused primarily in two directions: (i) further testing of the vaccine potency of paramyosin against schistosomes and other parasitic worms; and (ii) characterisation of the protein at the molecular and biochemical levels.

Schistosoma mansoni induces the synthesis of IL-6 in pulmonary microvascular endothelial cells: role of IL-6 in the control of lung eosinophilia during infection.

The nature of the interactions between the intravascular parasite Schistosoma mansoni and the host pulmonary vasculature is critical in determining the outcome of infection. In this report, we show that lung schistosomula selectively induce the synthesis of IL-6 mRNA and protein in cultured human and mouse lung microvascular endothelial cells (EC) and that parasite excretory/secretory lipophilic compounds, particularly prostaglandin E(2), are responsible for this effect. In vivo, a striking increase of IL-6 expression is observed in the pulmonary microvasculature of S. mansoni-infected C57BL/6 mice suggesting that, in vivo, parasites also induce the synthesis of IL-6 in lung EC. In infected mice, IL-6 deficiency results in an accelerated mobilization of eosinophils into the lung tissue and in a dramatic increased number of recruited leukocytes, particularly eosinophils, in the airway. This effect is associated with an enhanced production of eotaxin (CCL11) and IL-5 in the lungs of IL-6 knockout (KO) animals. Finally, compared to wild-type mice, we detect a dramatic increased level of parasite mortality in the lungs of IL-6 KO mice. Taken together, we suggest that parasite larvae activate EC to produce IL-6 to escape the inflammatory reaction that develops in the lungs of infected hosts. Finally, we show that the parasite-induced IL-6 synthesis is mediated by a protein kinase A-dependent pathway that principally targets the cAMP-response element and the nuclear factor-kappaB sites from the -256/+20 region of the IL-6 promoter.

Survival of bird schistosomes in mammalian lungs

Bird schistosome cercariae have a low specificity to vertebrate skin and, thus, they are also able to penetrate into mammals. As a consequence, a hypersensitive skin response—cercarial dermatitis—develops. It was thought that the parasites die in the skin soon after penetration. Our results on Trichobilharzia szidati and Bilharziella polonica in the non-specific murine host confirm that some of the penetrating bird schistosomes may fully transform to schistosomula and migrate to the lungs. They persist there for up to 10 days post exposure. In a duck, the worms grow and feed rapidly, but in a mouse the lung schistosomula seem to be inhibited in their development. However, TEM results show that there is no damage to the tegument of these larvae and no immune effector cells attack the parasites. These results suggest that the parasite's failure in the murine host might be caused by some immunologically unrelated factors.

Proteosome delivery of a protective 9B-antigen against Schistosoma mansoni

We have previously characterized a stage specific, partially protective protein denoted 9B-antigen. This antigen is of 450 kDa in its native form but upon SDS–PAGE in reducing conditions it exhibits two subunits of 30 kDa and 45 kDa. The 9B-antigen is localized at the surface of schistosomula and persists at the surface of lung schistosomula. The 9B-antigen is also localized in internal organs of a vital function in the parasite such as flame cells and cytoplasmic tubes. Infected individuals or mice vaccinated with irradiated cercariae recognize the 9B-antigen. We have previously shown that when injected with complete Freunds adjuvant, the 9B-antigen can induce 40% protection against challenge infection. In this study we have used a more effective delivery system for this antigen. The 9B-antigen was coupled to proteosomes derived from meningoccocal outer membrane proteins. Vaccination of mice with this complex increased the protection level to 60%. Sera from these vaccinated mice induced high levels of complement mediated cytotoxicity of the parasite. Since the proteosomes are approved for human use, these results are promising towards the development of a vaccine against schistosomiasis.

Recruitment of lymphocytes to the lung through vaccination enhances the immunity of mice exposed to irradiated schistosomes.

The effector mechanism, which operates against challenge parasites in the lungs of C57BL/6 mice vaccinated once with irradiated cercariae of Schistosoma mansoni, is mediated by CD4+ T helper lymphocytes. However, adoptive transfer of immunity from vaccinated donors to naive recipients by using sensitized T cells has not proved successful. One explanation may be that the recruitment of sensitized T lymphocytes to the lungs by vaccinating parasites to arm that organ is not reproduced by transfer protocols. We have used the technique of parabiosis, as a means of adoptive transfer, to demonstrate the relevance of pulmonary T cells to protection. Sensitized and naive partners were joined surgically for a 28-day period, coincident with priming of the immune system. A vascular union rapidly developed, and sensitized T cells were detected in the spleens of the naive partners. When parabionts were challenged percutaneously 10 days after separation, the level of immunity transferred to the naive partners was approximately two-thirds that of their vaccinated counterparts. The naive partners, unlike the vaccinated animals, did not recruit lymphocytes to the lungs during the priming period. In contrast, after percutaneous challenge, schistosome-specific lymphocytes were recruited to the lungs of both separated parabionts. The importance of lymphocytes recruited to the lungs during the primary response was revealed by an intravenous challenge with lung schistosomula; this eliminates the opportunity for secondary immune responses prior to parasite arrival in the lungs. In this situation, the vaccinated partners showed 47% immunity while the naive partners were not protected. We conclude that the presence of specific T cells in the lungs at the time of challenge confers a significant advantage, permitting a more effective recall response than in animals lacking such resident cells.

Schistosoma japonicum: immunolocalization of paramyosin during development.

This paper describes the localization of paramyosin immunoreactivity in Schistosoma japonicum and represents the first comparative immunolocalization study among schistosome adult, cercariae and lung schistosomula by electron microscopy. A polyclonal antibody was utilized to immunolabel paramyosin or paramyosin-like proteins. Paramyosin was localized within the muscle layer of all 3 developmental stages. Furthermore, paramyosin was localized within granules of the post-acetabular glands of cercariae, and within the tegument matrix and surface of lung schistosomules. Adults and cercariae did not display any detectable paramyosin on the surface or within the tegument. The possible functions of paramyosin within S. japonicum and the relevance of these findings in relation to the reported protective properties of paramyosin as an anti-schistosome vaccine target molecule are discussed.

Comparison of Two Strains of Schistosoma mansoni with Respect to the Sites and Kinetics of Immune Elimination in Irradiated Cercaria-Immunized Mice

A comparison was made of the sites of elimination of NMRI and Mill Hill strains of Schistosoma mansoni in C57BL/6J mice previously immunized with 50 krad gamma-irradiated cercariae of the homologous strain. In the first experiment, the fate of percutaneous challenge infections with 75Se-labeled cercariae was evaluated by autoradiography of tissue squashes and hepatic portal perfusion. For both strains of parasite, migration from skin to lungs was delayed but not reduced in immunized mice relative to controls, with immune elimination taking place at some point after migration to the lungs. In a second experiment, resistance to the Mill Hill strain of S. mansoni was compared in mice challenged by percutaneous infection with cercariae and by intravenous injection with lung schistosomula. Both types of challenge were shown to be vulnerable to immune elimination. We conclude that under the conditions employed in this study, there is no significant difference between the NMRI and Mill Hill strains of S. mansoni in the patterns of migration and elimination, with most or all elimination in both control and immunized mice taking place after migration from the skin.

Localization of the A1.12/9 antigen family to the neurones, putative sensory receptors and tegument of Schistosoma mansoni

The A1.12/9 antigen family were localized to the periphery of vesicle-like structures in neurones and in the putative sensory receptors of cercariae by immuno-electron microscopy. After transformation into schistosomula, the antigens were rapidly lost until, after 18 h, no immunoreactivity could be detected. Expression resumed at approximately 36 h post-transformation and had returned to high levels by 4 days post-infection. This level was maintained through to the adult worm stage. In 4 day lung schistosomula and in 14 day and 21 day liver schistosomula immunolabelling was observed within the matrix of the tegument itself and also in association with the tegumental membrane and vesicles within the tegument. These properties suggest that the A1.12/9 antigens may be involved in neuropeptide processing pathways. In particular, this family may represent the schistosome homologue of the granin family with which they share common properties and some sequence homology.

Evidence that both normal and immune elimination of Schistosoma mansoni take place at the lung stage of migration prior to parasite death.

The number and distribution of autoradiographic foci observed in this and previous studies following percutaneous infection with 75Se-labeled Schistosoma mansoni cercariae indicate that the lungs are the principal site of worm elimination in both normal mice and mice immunized with irradiated cercariae. It was observed in the present study, however, that the intensities of the autoradiographic foci produced in the lungs during both the normal (early) and immune (late) phases of elimination were identical to those of foci produced in the livers of the same mice by larvae shown to be alive. In contrast, foci produced in the lungs by heat-killed, intravenously injected, lung schistosomula became smaller and fainter with time, disappearing completely between seven and 10 days after injection in normal mice and between four and six days in immunized mice. These results indicate that although the targets of both normal and immune elimination do not proceed beyond the lung stage of migration, they do not die in the lungs. A possible explanation for this paradoxical situation, for which there is some experimental evidence, is that unsuccessful migrators leave the blood stream, enter alveoli, pass up the trachea, and are eventually digested in the gastrointestinal tract or eliminated from the body intact.

Temporal variation in the carbohydrate and peptide surface epitopes in antibody‐dependent, eosinophil‐mediated killing of Schistosoma mansoni schistosomula

Changes in the surface antigenicity and susceptibility to eosinophil-dependent killing during in vitro development of schistosomula of Schistosoma mansoni, were examined using sera from rabbits and mice immunized with antigens that are shed from the schistosomulum in vitro (shed antigen), a carbohydrate extract of shed antigen (SAg/CHO) or a periodate-insensitive fraction of shed antigen (SAg/PEP). Anti-SAg/CHO antisera recognised mainly carbohydrate epitopes on the parasite surface, whilst anti-SAg/PEP antisera bound to periodate-insensitive, putative peptide, surface epitopes. Anti-SAg/PEP antibodies failed to recognise the surface of newly transformed schistosomula unless the parasite was first treated with sodium periodate, suggesting that these epitopes may be masked by periodate sensitive (i.e., carbohydrate) epitopes. There was an increase in anti-SAg/PEP antibody binding to the larval surface with age of the parasite in vitro; five-day-old lung schistosomula were also recognised by anti-SAg/PEP antisera. In contrast, anti-SAg/CHO antibody binding declined with parasite age, and failed totally to recognise lung schistosomula. This change in epitope expression was reflected in eosinophil-dependent cytotoxicity assays, with anti-SAg/CHO antisera killing young larvae and anti-SAg/PEP antisera only killing older larvae. Lung worms were not killed by either antisera. The difference in epitopes recognised by the antisera was also reflected in the antigens identified by immunoprecipitation and SDS-PAGE.

Schistosoma japonicum (Chinese strain): characterization of two monoclonal antibodies which recognized common epitopes in the tegument of adult worms and schistosomula

Two monoclonal antibodies have been produced that bind to the common epitopes on adult and schistosomula of S. japonicum (Chinese strain). The isotypes of these monoclonal antibodies were determined to be IgM (designated 8G9-5) and IgG2a (designed 9E7) respectively. The target epitopes of 8G9-5 and 9E7, as analysed by immunoblotting assay, are at Mr of 64 kDa and 54 kDa respectively. Indirect immunofluorescence assay using frozen sections of adult worms, intact mechanically transformed schistosomula and 5-day-old lung schistosomula indicated that both epitopes are located in the tegument of both stages of parasite. These epitopes appear to be two of several major immunogenic proteins on the tegument of both young and adult worms.