Irradiated Cercariae Research Articles

Micro Array-Assisted Analysis of Anti-Schistosome Glycan Antibodies Elicited by Protective Vaccination With Irradiated Cercariae.

Baboons vaccinated with radiation-attenuated cercariae develop high levels of protection against schistosome infection, correlating to high antibody titres towards schistosome antigens with unknown molecular identity. Using a microarray consisting of glycans isolated from different life-stages of schistosomes, we studied the anti-glycan immunoglobulin (Ig) G and IgM responses in vaccinated and challenged baboons over a time course of 25 weeks. Anti-glycan IgM responses developed early after vaccination, but did not rise in response to later vaccinations. In contrast, anti-glycan IgG developed more slowly, but was boosted by all five subsequent vaccinations. High IgM and IgG levels against O-glycans and glycosphingolipid glycans of cercariae were observed. At the time of challenge, while most antibody levels decreased in the absence of vaccination, IgG towards a subset of glycans containing multiple-fucosylated motifs remained high until 6 weeks post-challenge during challenge parasite elimination, suggesting a possible role of this IgG in protection.

Induction of Protective Immune Responses Against Schistosomiasis haematobium in Hamsters and Mice Using Cysteine Peptidase-Based Vaccine.

One of the major lessons we learned from the radiation-attenuated cercariae vaccine studies is that protective immunity against schistosomiasis is dependent on the induction of T helper (Th)1-/Th2-related immune responses. Since most schistosome larval and adult-worm-derived molecules used for vaccination uniformly induce a polarized Th1 response, it was essential to include a type 2 immune response-inducing molecule, such as cysteine peptidases, in the vaccine formula. Here, we demonstrate that a single subcutaneous injection of Syrian hamsters with 200 μg active papain, 1 h before percutaneous exposure to 150 cercariae of Schistosoma haematobium, led to highly significant (P < 0.005) reduction of >50% in worm burden and worm egg counts in intestine. Immunization of hamsters with 20 μg recombinant glyceraldehyde 3-phosphate dehydrogenase (rSG3PDH) and 20 μg 2-cys peroxiredoxin-derived peptide in a multiple antigen peptide construct (PRX MAP) together with papain (20 μg/hamster), as adjuvant led to considerable (64%) protection against challenge S. haematobium infection, similar to the levels reported with irradiated cercariae. Cysteine peptidases-based vaccination was also effective in protecting outbred mice against a percutaneous challenge infection with S. haematobium cercariae. In two experiments, a mixture of Schistosoma mansoni cathepsin B1 (SmCB1) and Fasciola hepatica cathepsin L1 (FhCL1) led to highly significant (P < 0.005) reduction of 70% in challenge S. haematobium worm burden and 60% reduction in liver egg counts. Mice vaccinated with SmCB1/FhCL1/rSG3PDH mixture and challenged with S. haematobium cercariae 3 weeks after the second immunization displayed highly significant (P < 0.005) reduction of 72% in challenge worm burden and no eggs in liver of 8–10 mice/group, as compared to unimmunized mice, associated with production of a mixture of type 1- and type 2-related cytokines and antibody responses.

A Meta-Analysis of Experimental Studies of Attenuated Schistosoma mansoni Vaccines in the Mouse Model.

Schistosomiasis is a water-borne, parasitic disease of major public health importance. There has been considerable effort for several decades toward the development of a vaccine against the disease. Numerous mouse experimental studies using attenuated Schistosoma mansoni parasites for vaccination have been published since 1960s. However, to date, there has been no systematic review or meta-analysis of these data. The aim of this study is to identify measurable experimental conditions that affect the level of protection against re-infection with S. mansoni in mice vaccinated with radiation attenuated cercariae. Following a systematic review, a total of 755 observations were extracted from 105 articles (published 1963–2007) meeting the searching criteria. Random effects meta-regression models were used to identify the influential predictors. Three predictors were found to have statistically significant effects on the level of protection from vaccination: increasing numbers of immunizing parasites had a positive effect on fraction of protection whereas increasing radiation dose and time to challenge infection had negative effects. Models showed that the irradiated cercariae vaccine has the potential to achieve protection as high as 78% with a single dose vaccination. This declines slowly over time but remains high for at least 8 months after the last immunization. These findings provide insights into the optimal delivery of attenuated parasite vaccination and into the nature and development of protective vaccine induced immunity against schistosomiasis, which may inform the formulation of human vaccines and the predicted duration of protection and thus frequency of booster vaccines.

Effect of Ultra Violet Attenuated Cercariae on Antibody Response in Rabbits

Ultraviolet (UV) light is an electromagnetic radiation below the visible wavelengths. Their use in cercariae attenuation needs to be examined. In the present work, Schistosoma mansoni cercariae was exposed to UV radiation. Few studies showed previously an inflicted damage seen on the adult schistosome worms developed from irradiated cercariae. The aim of the study was to find out whether this damage was attributed to direct effect of UV-irradiation on cercariae or due to the host's immunogenicity induced by UV- irradiated cercariae. Single as well as multiple exposures of cercariae to UV Light were examined. They were exposed to UV light, for 1, 2 and 3 hrs and then, were subjected to different treatments: the first one was for the assessment of cercarial viability after one hour of the 3 different treatments. The second one was for rabbit infection and the third one was for cercarial antigen preparation. The cercarial antigens (Ag) were recognized by protective antibodies (IgG1 fractions) which then, were separated and purified from vaccinated rabbit's serum. This cercarial Ag was identified as UVISmC1 gene encoding a protein showing 100% identity at the amino acid level with previously identified S. mansoni clones; theses clones are encoding 51.7 kDa antigens elicited as a result of direct effect of UV radiation on cercariae as well as host's immunogenicity induced by UV irradiated cercariae. This was verified by the recognition of this Ag prepared from E coli clones isolated from cDNA expression library. In conclusion, the data showed a remarkable potency of the UV-radiation-attenuated cercaria in eliciting differential high effectiveness in Ab response under laboratory conditions at 1, 2 and 3 hrs of UV light output at 254 nm. An understanding of the protective immune response elicited by RA cercaria may help in designing a candidate vaccine which is still needed.

A HIGH POTENT ANTIBODY RESPONSE IN RABBITS INFECTED WITH ULTRA VIOLET ATTENUATED CERCARIAE

Ultraviolet (UV) light is an electromagnetic radiation below the visible wavelengths. Their use in cercariae attenuation needs to be examined. In the present work, Schistosoma mansoni cercariae was exposed to UV radiation. Few studies showed previously an inflicted damage seen on the adult schistosome worms developed from irradiated cercariae. The aim of the study was to find out whether this damage was attributed to direct effect of UV-irradiation on cercariae or due to the host's immunogenicity induced by UV- irradiated cercariae. Single as well as multiple exposures of cercariae to UV Light were examined. They were exposed to UV light, for 1, 2 and 3 hrs and then, were subjected to different treatments: the first one was for the assessment of cercarial viability after one hour of the 3 different treatments. The second one was for rabbit infection and the third one was for cercarial antigen preparation. The cercarial antigens (Ag) were recognized by protective antibodies (IgG1 fractions) which then, were separated and purified from vaccinated rabbit's serum. This cercarial Ag was identified as UVISmC1 gene encoding a protein showing 100% identity at the amino acid level with previously identified S. mansoni clones; theses clones are encoding 51.7 kDa antigens elicited as a result of direct effect of UV radiation on cercariae as well as host's immunogenicity induced by UV irradiated cercariae. This was verified by the recognition of this Ag prepared from E coli clones isolated from cDNA expression library. In conclusion, the data showed a remarkable potency of the UV-radiation-attenuated cercaria in eliciting differential high effectiveness in Ab response under laboratory conditions at 1, 2 and 3 hrs of UV light output at 254 nm. An understanding of the protective immune response elicited by RA cercaria may help in designing a candidate vaccine which is still needed.

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.'

Artemether Treatment of Prepatent Schistosoma japonicum Induces Resistance to Reinfection in Association with Reduced Pathology

Artemether (ART) is a well-described antimalarial with efficacy against juvenile schistosomes, with 7-day-old schistosomula being particularly susceptible. Both ART-affected worms and parasites developing from irradiated cercariae die at similar times after infection. Our aim was to determine if ART treatment of prepatent schistosomiasis japonica may result in the generation of a protective immune response. Female CBA mice were treated with a single dose of ART at defined time points after percutaneous infection with a virulent Chinese mainland strain of Schistosoma japonicum. Half of the mouse cohorts were subjected to homologous parasite strain reinfection after drug treatment to assess protective effects of ART therapy. Two independent trials demonstrated that a statistically significant (58% and 50%) reduction in challenge worm burden occurred after reinfection of those mice treated with ART at two weeks p.i. A reduction in the IL-4 response to soluble worm antigen preparation (SWAP) was also seen in ART-treated mice but with no correlation to reinfection resistance. In the Chinese mainland strain used, ART treatment of prepatent infection at the appropriate time point induced resistance to reinfection. There was also an anti-pathology effect observed in ART-treated mice that remained significant after reinfection.

Altered Patterns of Gene Expression Underlying the Enhanced Immunogenicity of Radiation-Attenuated Schistosomes

BackgroundSchistosome cercariae only elicit high levels of protective immunity against a challenge infection if they are optimally attenuated by exposure to ionising radiation that truncates their migration in the lungs. However, the underlying molecular mechanisms responsible for the altered phenotype of the irradiated parasite that primes for protection have yet to be identified.Methodology/Principal FindingsWe have used a custom microarray comprising probes derived from lung-stage parasites to compare patterns of gene expression in schistosomula derived from normal and irradiated cercariae. These were transformed in vitro and cultured for four, seven, and ten days to correspond in development to the priming parasites, before RNA extraction. At these late times after the radiation insult, transcript suppression was the principal feature of the irradiated larvae. Individual gene analysis revealed that only seven were significantly down-regulated in the irradiated versus normal larvae at the three time-points; notably, four of the protein products are present in the tegument or associated with its membranes, perhaps indicating a perturbed function. Grouping of transcripts using Gene Ontology (GO) and subsequent Gene Set Enrichment Analysis (GSEA) proved more informative in teasing out subtle differences. Deficiencies in signalling pathways involving G-protein–coupled receptors suggest the parasite is less able to sense its environment. Reduction of cytoskeleton transcripts could indicate compromised structure which, coupled with a paucity of neuroreceptor transcripts, may mean the parasite is also unable to respond correctly to external stimuli.Conclusions/SignificanceThe transcriptional differences observed are concordant with the known extended transit of attenuated parasites through skin-draining lymph nodes and the lungs: prolonged priming of the immune system by the parasite, rather than over-expression of novel antigens, could thus explain the efficacy of the irradiated vaccine.

The development of hepatic granulomas in 20 Krad irradiated Schistosoma mansoni cercaria vaccinated grivet monkeys ( Cercopithecus aethiops aethiops)

The absence of appropriate animal models is a challenge for those designing anti-schistosome vaccines. In order to observe the development of hepatic granulomata, 20 Krad irradiated cercaria vaccinated grivet monkeys ( Cercopithecus aethiops aethiops) were experimentally infected with Schistosoma mansoni. Most human clinical manifestations of schistosomiasis were noted in the infected monkeys. Intraperitonial immunization with 20 Krad irradiated cercaria resulted in a relatively less granulomatous reaction and lower egg mass ( P < 0.05) than in the controls. The mean total adult worms and egg production was also lower in the vaccinated group. This study underlines the significant role of irradiated cercariae vaccination on the establishment of female adults and consequent reduction of pathology in the grivet Cercopithecus aethiops aethiops.

Serum cytokines of the 20 Krad-irradiated S. mansoni cercariae vaccinated, primary and superinfected Cercopethicus aethiops aethiops

Appropriate animal models are necessary to better understand the immune response in schistosomiasis. Schistosoma mansoni infection was established using irradiated cercariae in Cercopithecus aethiops aethiops ( Grivet monkey) to describe immune responses of the serum cytokines, IL-4, IL-10, IL-12, IFN- γ, and TNF- α. Intraperitoneal irradiated cercariae immunization on three occasions resulted in some differences of cytokine production. In primary infection, IL-4 was significantly raised ( p = 0.03) in the immunized monkeys, and there was an insignificant increase ( p > 0.05) in IL-10. However, ova excretion did not influence the cytokines, except in the controls where both IL-4 and IL-10 were significantly increased ( p < 0.05). In the controls, IL-12 and INF-γ levels were lower after ova excretion, but the inflammatory TNF-α increased ( p = 0.049) and these findings can be associated with more liver pathogenesis in the group. Thus, this work has indicated the potential importance of anti-schistosome vaccine studies on the grivet monkeys.

Comparison of Schistosoma mansoni irradiated cercariae and Sm23 DNA vaccines

Immunization with defined antigens is generally less effective at inducing host protection against experimental infection with Schistosoma mansoni than vaccination with attenuated infective cercariae. We predicted that quantitative and/or qualitative differences existed between the immune responses generated to attenuated cercariae and those induced by defined antigens. Thus, we compared immune responses typically associated with protection in the murine model between animals vaccinated with attenuated cercariae and mice immunized with DNA encoding Sm23, a schistosome integral membrane protein that has previously been shown to confer protection. Mice vaccinated three times with attenuated cercariae demonstrated higher levels of protection than Sm23-vaccinated animals but spleen cells from Sm23 DNA vaccinated mice produced significantly higher levels of schistosome antigen-specific IFN-gamma. Both vaccines induced similar levels of Sm23-specific antibody and post-challenge dermal inflammation. However, the pulmonary inflammatory responses following challenge were much less pronounced in DNA immunized animals compared to those receiving irradiated cercariae. Thus, although Sm23 DNA vaccination effectively induced parasite-specific IFN-gamma and antibody responses, it failed to evoke other critical responses needed for optimal vaccine efficacy.

TRIGGERING OF HIGH-LEVEL RESISTANCE AGAINST SCHISTOSOMA MANSONI REINFECTION BY ARTEMETHER IN THE MOUSE MODEL

Artemether, a methyl ether derivative of dihydroartemisinin, not only exhibits antimalarial properties, but also possesses strong activity against schistosomula, the immature stages of a parasitic worm that can cause schistosomiasis. To test if the effect would be similar to that of irradiation with respect to the induction of immunologic protective responses, groups of mice were infected with Schistosoma mansoni cercariae and treated with artemether at 1-3 weeks post-infection. Control mice were either infected with normal cercariae or with cercariae exposed to radiation that permitted early development but not maturation of the parasites. The mice were challenged six weeks after the initial infection, and the mean numbers of schistosomes recovered in the various groups were calculated upon dissection eight weeks post-challenge. The administration of artemether two weeks after the initial infection resulted in 58% protection, while giving the drug three weeks post-infection increased the level of protection to 81%. This level of protection is as high as that normally obtained by immunization with irradiated cercariae (84% in the present study) and is superior to the level of resistance obtained with any individual schistosome vaccine candidate antigen thus far reported.

Schistosoma mansoni: antigen-presenting cells emigrating from skin exposed to attenuated cercariae activate lymphoid cells and transfer protection in C57B1/6 mice.

C57B1/6 mice develop significant levels of protection to a challenge infection after percutaneous exposure to irradiated Schistosoma mansoni cercariae. Although some circumstantial evidence has suggested that antigen-presenting cells (APCs) within the skin play a role in priming anti-schistosomula effector mechanisms, no direct evidence has been presented. In this study, we describe efforts to directly test whether skin-resident APCs exposed to irradiated cercariae are capable of mediating responses consistent with previously proposed mechanisms associated with delayed-type hypersensitivity reactions. We demonstrate that a population of APCs emigrates from the skin after percutaneous vaccination and that these cells are able to induce proliferation of S. mansoni-specific lymphocytes. We describe our experiments conducted to confirm that proliferation is dependent on major histocompatibility complex (MHC) Class-II interactions and cell-to-cell contact between APCs and lymphocytes. Immunohistological staining of emigrating cells revealed a population of large MHC Class-II+ cells with a morphology characteristic of mature dendritic cells. On recovery and adoptive transfer into naive mice, these cells demonstrated the ability to mediate protection to a challenge infection at levels similar to those in percutaneously vaccinated controls. This confirms that cutaneous APCs can initiate anti-schistosomula effector mechanisms in C57B1/6 mice after percutaneous vaccination.

Dominant antibody responses to Fucα1-3GalNAc and Fucα1-2Fucα1-3GlcNAc containing carbohydrate epitopes in Pan troglodytes vaccinated and infected with Schistosoma mansoni

The development of the humoral anti-glycan immune response of chimpanzees, either or not vaccinated with radiation-attenuated Schistosoma mansoni cercariae, was followed during 1 year after infection with S. mansoni. During the acute phase of infection both the vaccinated and the control chimpanzees produce high levels of immunoglobulin G (IgG) antibodies against carbohydrate structures that are characteristic for schistosomes carrying the Fucα1-3GalNAc and Fucα1-2Fucα1-3GlcNAc motifs, but not to the more widespread occurring structures GalNAcβ1-4GlcNAc, GalNAcβ1-4(Fucα1-3)GlcNAc, and Galβ1-4(Fucα1-3)GlcNAc (Lewis x). In addition, high levels of IgM antibodies were found against the trimeric Lewis x epitope. Apparently, the schistosome-characteristic carbohydrate structures are dominant epitopes in the anti-glycan humoral immune response of the chimpanzees. All chimpanzees showed an increase in the level of antibodies against most of the carbohydrate structures tested directly after vaccination, peaking at challenge time and during the acute phase of infection. With the exception of anti-F-LDN antibody responses, the anti-carbohydrate antibody responses upon schistosome infection of the vaccinated animals were muted in comparison to the control animals. Index Descriptors and Abbreviations: Anti-carbohydrate antibodies, glycoconjugates, humoral immune response, infection, Schistosoma mansoni, synthetic glycan epitopes, trematode, vaccination, irradiated cercariae. Circulating cathodic antigen (CCA), Fucα1-3GalNAc(Fucα1-3) 0/1GlcNAcαGal (F-LDN(-F) 0/1), goat anti-human IgG (GaHuIgG), goat anti-human IgM (GaHuIgM), Hepes-buffered saline (HBS), immunoglobulin (Ig), keyhole limpet haemocyanin (KLH), GalNAcβ1-4GlcNAc (LDN), GalNAcβ1-4(Fucα1-3)GlcNAc (LDN-F), GalNAcβ1-4(Fucα1-2Fucα1-3)GlcNAc (LDN-DF), Galβ1-4(Fucα1-3)GlcNAc (Lewis x), peripheral blood mononuclear cells (PBMCs), response units (RUs), surface plasmon resonance (SPR).

Cloning and characterization of a Schistosoma mansoni 1H and 30S clones as two tegumental vaccine candidate antigens.

Two Schistosoma mansoni cDNA clones 30S and 1H were identified by immunoscreening of sporocyst lambdagt11 library and by random sequencing of clones from lambdaZap libraries, respectively. Clone 30S was one of 30 clones identified by an antibody raised against tegument of 3-h schistosomules. The clone was found to encode an 81 amino-acid protein fragment. It was expressed in Escherichia coli as a fusion protein of calculated molecular mass of about 35 kDa with C-terminus of Schistosoma japonicum glutathione-S-transferase (Sj26; about 26 kDa). The recombinant fusion protein was specifically recognized by serum of rabbits immunized with irradiated cercariae. Clone 1H is one of 76 expressed sequence tags derived from an adult worm library. It encodes the complete sequence of a tegumental membrane protein, Sm13. The 104 amino-acid open reading frame encodes a protein with a calculated molecular mass of about 11.9 kDa. Clone 1H was expressed in E. coli as an insoluble fusion protein with Sj26 of about 40 kDa. In Western blots, the fusion protein was recognized by serum from rabbits vaccinated with irradiated cercariae but not by preimmune rabbit sera. The cloning, characterization and expression of those proteins are therefore potentially usefull for vaccine development.

Cloning and characterization of a Schistosoma mansoni 1H and 30S clones as two tegumental vaccine candidate antigens.

Two Schistosoma mansoni cDNA clones 30S and 1H were identified by immunoscreening of sporocyst lambdagt11 library and by random sequencing of clones from lambdaZap libraries, respectively. Clone 30S was one of 30 clones identified by an antibody raised against tegument of 3-h schistosomules. The clone was found to encode an 81 amino-acid protein fragment. It was expressed in Escherichia coli as a fusion protein of calculated molecular mass of about 35 kDa with C-terminus of Schistosoma japonicum glutathione-S-transferase (Sj26; about 26 kDa). The recombinant fusion protein was specifically recognized by serum of rabbits immunized with irradiated cercariae. Clone 1H is one of 76 expressed sequence tags derived from an adult worm library. It encodes the complete sequence of a tegumental membrane protein, Sm13. The 104 amino-acid open reading frame encodes a protein with a calculated molecular mass of about 11.9 kDa. Clone 1H was expressed in E. coli as an insoluble fusion protein with Sj26 of about 40 kDa. In Western blots, the fusion protein was recognized by serum from rabbits vaccinated with irradiated cercariae but not by preimmune rabbit sera. The cloning, characterization and expression of those proteins are therefore potentially usefull for vaccine development.

Comparison of the vaccine efficacy of γ-irradiated Schistosoma japonicum cercariae with the defined antigen Sj62(IrV-5) in pigs

Development of a vaccine against Schistosoma japonicum which can protect both man and the domestic animal zoonotic reservoirs of infection would be an invaluable tool in attempts to control this infection in those areas in which conventional control methods have failed to break transmission. The pig is a natural host of S. japonicum and because of its anatomical and immunological similarities to humans, it is a potentially valuable host for studies on S. japonicum in particular and schistosomes in general. Radiation-attenuated cercariae are highly effective in inducing immunity in experimental schistosomosis and there are promising reports of partial protection against schistosomes with recombinant-derived individual antigens. In the present study we have set out to establish a protocol for inducing protection with γ-irradiated cercariae in pigs and to assess the protective capacity of recombinant and naked DNA formulations of Sj62, a 62 kDa region of S. japonicum myosin. The corresponding S. mansoni version or Sj62, recombinant IrV-5, has previously been implicated in irradiated vaccine immunity in S. mansoni infections and has been shown to induce high levels of immunity in a variety of hosts. Groups of pigs were immunised three times at 2-week intervals with 2000 cercariae irradiated at 20 krad, with Sj62 as a recombinant (rSj62) incorporated in Freund’s adjuvant, a micellar preparation, or as a naked DNA construct. Vaccination with irradiated cercariae did not induce significant anti-Sj62 antibody but following intramuscular challenge with 2000 cercariae, the vaccinated pigs showed >95% resistance as assessed by reduced faecal egg output, worm tissue egg burdens and also reduced septal fibrosis. Immunisation with each of the Sj62 formulations induced significant anti-Sj62 antibody responses, the highest titre (>12,800) being with the Freund’s preparation but none of the Sj62-immunised groups showed significant resistance to challenge. The data suggest that Sj62 shows little promise as a vaccine candidate for schistosomosis.

Characterization, cloning and immunogenicity of antigens released by transforming cercariae of Schistosoma mansoni.

A schistosome infection is initiated when the parasite penetrates the skin of a susceptible host. Relatively large quantities of protein are released by transforming cercariae compared to later larval stages. This represents the first parasite material to which the host's immune system is exposed, yet little is known about the proteins which are released during the first few hours post-transformation. We have shown that antiserum raised against such molecules was capable of imparting protection against a schistosome challenge infection upon passive transfer to naïve mice. By screening a cercarial cDNA library with this serum, 38 positive clones were identified. Sequence analysis showed these to represent 8 different molecules which included Schistosoma mansoni 21-7 kDa antigen, calcium-binding-protein and the vaccine candidate glutathione S-transferase (Sm28GST). In addition, 5 clones were isolated, 1 of which had significant homology to many cytochrome C proteins, another with leukocyte elastase inhibitors and 3 which represented novel molecules. Four clones were expressed in a prokaryotic high-level expression vector, sera produced against each purified recombinant protein and used subsequently to probe Western blots and parasite sections. The leukocyte elastase inhibitor homologue and 2 unknowns induced significant proliferation by lymph node cells recovered from mice vaccinated with irradiated cercariae. More strikingly, the 2 novel proteins stimulated very high levels of interferon gamma (IFNgamma) secretion both by lymph node cells and those recovered by broncho-alveolar lavage from the lungs of vaccinated mice. Such results will be discussed in the context of vaccine development.

Schistosomiasis Vaccines: A Response to a Devils’ Advocate’s View

work, even on non-human primates, will not improve our knowledge of human protective immune reactions. While humans should not be subjected to harmful experimentation, it is equally important not to delay efforts to alleviate the problem of schistosomiasis in endemic areas. We feel that schistosomiasis cannot be defeated with chemotherapy alone, but rather than arguing the advantages of drugs versus vaccines, we suggest that these approaches are not mutually exclusive, as drugs provide short-term reduction of morbidity, while vaccines hold the promise of long-term prevention. The development of a human vaccine is a long-term endeavour, but there are reasons to be optimistic. The strong protection reached with irradiated cercariae in experimental animals is the proof of principle, while practice is supported by studies showing human protective immune responses resulting in various degrees of resistance to infection. Gryseels feels that antigens eliciting natural immune responses are unlikely vaccine candidates and remains unimpressed by the notion of protective immunity. His first objection may well be correct, but it is also possible that the immune system is modulated in some general way, or by the schistosomes

Schistosoma mansoni phosphofructokinase: immunolocalization in the tegument and immunogenicity.

Schistosoma mansoni depends for its survival on glycolysis. Two glycolytic enzymes, glyceraldehyde-3P-dehydrogenase and triose-phosphate dehydrogenase, found in both the adult and schistosomular tegument, have been reported to confer partial protection against cercarial infection. This paper describes the immunogenic properties of phosphofructokinase (PFK), a rate-limiting enzyme of glycolysis, and its localization in the tegument and adjacent tissues. Recombinant schistosome PFK was used as antigen. A polyclonal antibody against purified PFK from Fasciola hepatica was affinity purified using recombinant PFK and used in combination with immunogold labelling to identify PFK by transmission electron microscopy in cryosections. In both adult worms and in schistosomula most immunogold label localized in the cytoplasmic syncytial region with less being found in the tegument. There was no significant PFK localization within or external to the outer membrane. Sera from mice immunized with recombinant S. mansoni PFK with Freund's adjuvant or alum plus rIL-12 demonstrated high titres of anti-PFK IgG, but no protection against cercarial infection. Sera from mice that were acutely or chronically infected or multiply exposed to irradiated cercariae did not recognize recombinant schistosome PFK in either Western blotting or ELISA. Similarly, sera from humans infected with S. mansoni did not recognize PFK. We conclude that in spite of the high immunogenicity of rPFK in mice, it is not a significant immunogen during the course of infection and does not confer protection from schistosomiasis. One main difference between PFK and the other 2 glycolytic enzymes seems to be the inaccessibility of PFK to the outside surface of the tegument.