Adult Worm Antigen Preparation Research Articles

Immunodiagnosis of cattle fascioliasis using a 27 kDa Fasciola gigantica antigen

Background and Aim:Diagnosis of fascioliasis depends on clinical symptoms and routine laboratory tests. Recently, antibodies and circulating antigens of Fasciola were used for detecting active infections. Therefore, this study aimed to identify Fasciola gigantica antigens in the sera of infected cattle using Western blotting and enzyme-linked immunosorbent assay (ELISA) for an accurate diagnosis of cattle infected with F. gigantica.Materials and Methods:Serum samples were obtained from 108, 23, and 19 cattle infected with Fasciola gigantica, Paramphistomum cervi, and Strongylids, respectively, including 57 non-infected cattle that were used as healthy cattle for the study. Western blotting and ELISA were then used to detect circulating Fasciola antigens at 27 kDa.Results:The target epitope was detected in an F. gigantica adult-worm antigen preparation, excretory/secretory products, and serum from cattle infected with F. gigantica. However, it was absent in sera from P. cervi, Strongylids, and healthy cattle. The purified 27 kDa F. gigantica (FPA-27) antigen was also detected in cattle serum using ELISA with high degrees of sensitivity and specificity (94% and 82%, respectively), and the area under the receiver operating characteristic curve was 0.89 with a highly significant correlation of p<0.0001.Conclusion:The FPA-27 is proposed to be a promising candidate for the serodiagnosis of fascioliasis in cattle.

Mycobacterium leprae–helminth co-infections and vitamin D deficiency as potential risk factors for leprosy: A case–control study in south-eastern Brazil

BackgroundEvidence suggests that biological mechanisms involved in helminth infections and vitamin deficiencies increase susceptibility to other infections. The aim of this study was to investigate the associations of helminth co-infection and select micronutrient deficiencies with leprosy using a case–control design. MethodsFrom 2016 to 2018, individuals aged ≥3 years were recruited at clinics in and around Governador Valadares, Minas Gerais, Brazil in three groups: cases of leprosy, household contacts and community-matched (non-contact) controls. Helminths were diagnosed through stool Kato Katz examination and serum reactivity to anti-soluble adult worm antigen preparation IgG4. Serum ferritin, 25-OH vitamin D and retinol concentrations were measured. Multi-variate logistic regression was conducted to identify associations with active leprosy. ResultsSeventy-nine cases of leprosy, 96 household contacts and 81 non-contact controls were recruited; 48.1% of participants were male with a median age of 40 years. Helminths were found in 7.1% of participants on Kato Katz test, all but one of which were Schistosoma mansoni, and 32.3% of participants were positive for S. mansoni serology. On multi-variate analysis, cases were more likely to be infected with helminths (diagnosed by stool) than household contacts [adjusted odds ratio (aOR) 8.69, 95% confidence interval (CI) 1.50–50.51]. Vitamin D deficiency was common, and was more likely in cases compared with non-contact controls (aOR 4.66, 95% CI 1.42,–15.33). Iron deficiency was not associated with leprosy, and vitamin A deficiency was not detected. ConclusionThese associations suggest that the immune consequences of schistosomiasis and vitamin D deficiency may increase the risk of active leprosy. Comorbid conditions of poverty deserve further study as addressing co-infections and nutritional deficiencies could be incorporated into programmes to improve leprosy control.

T cell-mediated immunity in CBA mice during Schistosoma japonicum infection

Characterisation of the cellular immune response to schistosomiasis is well established for Schistosoma mansoni but a comprehensive description of T cell-mediated immune responses against S. japonicum infection is lacking. Accordingly, 20 CBA mice were infected with cercariae of S. japonicum and the immune response at different time points was determined. Mouse spleen and liver lymphocytes were isolated from the mice and stimulated with schistosomal adult worm antigen preparation (SWAP) and schistosomal soluble egg antigen (SEA). There was a relatively higher Th1 immune response to SWAP compared to SEA at the early phase of infection (up to week 5 post challenge). However, a Th2 immune response directed against SEA was dominant at week 6 post-infection, a time point when the highest IgG response against both SWAP and, especially, SEA was generated. The regulatory immune response was highest at the early phase of the immune response (up to week 5 post challenge) followed by a rapid decline at week 6-post infection. Before egg-laying, S. japonicum induced a regulatory T cell immune response which may limit the early Th1-mediated immune response that is believed to be protective in murine schistosomiasis. Following egg laying, the immune response was polarized to a Th2 immune response mainly directed against the eggs and this may contribute to parasite survival.

A potential prophylactic strategy of anti-schistosomal immunization using nanotechnology in murine models

Background:The emergence of chemotherapy resistant schistosomes necessitates finding new prophylaxis strategies based on immunization by using a modified vaccine candidate. Objective: To assess thepotential of an anti-schistosomal vaccine modified by nanotechnology. Materials and Methods: Soluble adult worm antigen preparation (SWAP) either crude or/and loaded on chitosan nanoparticles (CSNPs)were used. The anti-schistosomal impact was assessed in terms of stools egg count,worm burden and ultrastructural studyof juvenile and adult schistosomes. Results:Chitosan nanoparticles potentiated SWAP efficacy to exceed the impact of crude SWAP.The combination of SWAP loaded on CSNPs and crude SWAP showed the best immunization effects regarding adult worm number andfecundity,as well as morphological and tegumental distortion of juvenile than adult stages. Conclusion:Modified SWAP on CSNPs gave better results in anti-schistosome immunization thancrude SWAP.Immunization by combining different antigens gave the best results.

B10 cells induced by Schistosoma japonicum soluble egg antigens modulated regulatory T cells and cytokine production of T cells.

A distinct subset of B cells, also known as regulatory B cells, can negatively regulate T cell immune responses, but the role of these cells in schistosomiasis has not been clarified. Soluble egg antigen (SEA) and soluble adult worm antigen preparation (SWAP), which are two important antigen sources during Schistosoma japonicum infection, both can induce Th1, Th2, Th17, and Treg cells and the corresponding cytokines. However, whether they can induce the production of regulatory B cells and the regulatory function of schistosome-induced regulatory B cells remains unclear. In our studies, we first analyzed the production of regulatory B cells stimulated by SEA or SWAP using flow cytometry and enzyme-linked immunosorbent assay, and observed these cells in mice immunized by SEA or SWAP. Then, B10 cells sorted by MicroBeads were co-cultured with CD4(+) T cells, and the proportion of Treg cells were detected. At the same time, the IFN-γ, IL-4, and IL-17 levels in the culture supernatant were measured. The results showed that B10 cells were preferentially induced by SEA in vitro, and B10 could also be induced in mice immunized by SEA. SEA-induced B10 cells promoted the expansion of regulatory T cells and induced IL-4 secretion, but inhibited IL-17 production. These findings reveal that the generation of B10 cells is determined by parasitic antigen, and suggest the function of B10 cell induced by SEA. This study significantly contributes to the understanding of the immune regulatory role in schistosomiasis and may help protect hosts from infection.

Association of Schistosoma mansoni-Specific IgG and IgE Antibody Production and Clinical Schistosomiasis Status in a Rural Area of Minas Gerais, Brazil

BackgroundStudies in murine models and human populations have indicated that the collagen-rich granulomatous response against parasite eggs trapped in the liver is associated with the development of severe hepatosplenic schistosomiasis, characterized by periportal fibrosis and portal hypertension. The role of the humoral response in parasite susceptibility has been well established, but its participation in disease severity remains poorly understood. In this work, we evaluated the relationship between parasite-reactive IgE and IgG levels and schistosomiasis morbidity in infected patients with similar parasite burdens.Methodology/Principal FindingsNinety-seven Schistosoma mansoni-infected individuals were subjected to clinical examination and abdominal ultrasound analysis. IgG reactivity and IgE concentration against Schistosoma mansoni soluble egg antigens (SEA) and adult worm antigen preparation (SWAP) were evaluated by ELISA assay. Multivariable linear regression models were used to evaluate the relationship between parasite-reactive antibodies and the co-variables investigated. The study population showed low parasite burden (median 30 eggs/g feces), constant re-infection, and signs of fibrosis was detected in more than 30% of individuals. Most infected individuals showed IgG reactivity, and the median concentrations of IgE anti-SEA and anti-SWAP antibodies were 1,870 and 1,375 ng/mL, respectively. There was no association between parasite burden and antibody response or any parameter of disease severity. However, IgG anti-SWAP level was positively associated with morbidity parameters, such as spleen size and thickness of portal vein at the entrance and secondary branch. In contrast, the data also revealed independent inverse correlations between concentration of parasite-reactive IgE and gallbladder wall thickness, a marker of fibrosis in schistosomiasis.Conclusions/SignificanceThe data indicate that IgG anti-SWAP is positively associated with severe schistosomiasis, independently of parasite burden, while high production of parasite-specific IgE is associated with mild disease in the human population. Antibody profiles are good correlates for schistosomiasis severity and could be tested as biomarkers of disease severity.

Adult worm‐specific IgE/IgG4 balance is associated with low infection levels of Schistosoma mansoni in an endemic area

Field studies have suggested an immune-mediated mechanism associated with resistance to Schistosoma mansoni infection. Overall, levels of specific IgE have been correlated with resistance to infection, whereas levels of IgG4 have been associated with susceptibility. This study aimed to evaluate serum levels of soluble adult worm antigen preparation (SWAP)-specific IgE and IgG4 in relation to current infection in a large casuistic of individuals living in an endemic area of schistosomiasis in Bahia, Brazil. The prevalence of S.mansoni infection was 37·7% and the mean parasite burden was 55·4 (0-2100) epg/faeces. There was no significant difference in the levels of SWAP-specific IgE in individuals with different parasite burden, whereas high producers of parasite-specific IgG4 presented higher parasite burden when compared to low IgG4 producers. Additionally, S.mansoni parasite load was positively correlated with the levels of specific IgG4 or total IgE. No significant correlation was observed between parasite burden and SWAP-specific IgE. Nevertheless, SWAP-specific IgE/IgG4 ratio was higher in uninfected or lightly infected individuals (1-99 epg/faeces) than in heavily infected ones (≥400 epg/feces). These findings highlight the important role of IgE/IgG4 ratio in the resistance to infection, which could be useful for further studies in schistosomiasis vaccine candidates.

Identification and Characterization of Argonaute Protein, Ago2 and Its Associated Small RNAs in Schistosoma japonicum

BackgroundThe complex life cycle of the genus Schistosoma drives the parasites to employ subtle developmentally dependent gene regulatory machineries. Small non-coding RNAs (sncRNAs) are essential gene regulatory factors that, through their impact on mRNA and genome stability, control stage-specific gene expression. Abundant sncRNAs have been identified in this genus. However, their functionally associated partners, Argonaute family proteins, which are the key components of the RNA-induced silencing complex (RISC), have not yet been fully explored.Methodology/Principal FindingsTwo monoclonal antibodies (mAbs) specific to Schistosoma japonicum Argonaute protein Ago2 (SjAgo2), but not SjAgo1 and SjAgo3, were generated. Soluble adult worm antigen preparation (SWAP) was subjected to immunoprecipitation with the mAbs and the captured SjAgo2 protein was subsequently confirmed by Western blot and mass spectrometry (MS) analysis. The small RNA population associated with native SjAgo2 in adult parasites was extracted from the immunoprecipitated complex and subjected to library construction. High-through-put sequencing of these libraries yielded a total of ≈50 million high-quality reads. Classification of these small RNAs showed that endogenous siRNAs (endo-siRNAs) generated from transposable elements (TEs), especially from the subclasses of LINE and LTR, were prominent. Further bioinformatics analysis revealed that siRNAs derived from ten types of well-defined retrotransposons were dramatically enriched in the SjAgo2-specific libraries compared to small RNA libraries constructed with total small RNAs from separated adult worms. These results suggest that a key function of SjAgo2 is to maintain genome stability through suppressing the activities of retrotransposons.Conclusions/SignificanceIn this study, we identified and characterized one of the three S. japonicum Argonautes, SjAgo2, and its associated small RNAs were found to be predominantly derived from particular classes of retrotransposons. Thus, a major function of SjAgo2 appears to associate with the maintenance of genome stability via suppression of retroelements. The data advance our understanding of the gene regulatory mechanisms in the blood fluke.

Protection Against Schistosoma Mansoni In Mice By using UV-Irradiation In Comparing With Soluble Adult Worm Antigen (SWAP)

Background: The present study aimed to investigate the dynamics of immunoregulatory factors implicated in the murine model of soluble adult worm antigen preparation (SWAP)and UV-irradiated adult worm extract-induced granulomatous hypo-responsivness. Material and Methods: Native male mice were classified into four groups. The first negative control (non-infected) group, the second positive control (infected) group, the third SWAP-vaccinated group and the fourth UV-irradiated adult worm extract-vaccinated group. The last two groups were injected subcutaneous by 0.2 ml (contained 100 µg protein) of the two used vaccines SWAP or UV-irradiated adult worm extract respectively, four weeks prior to infection. The 2nd, 3rd and 4th mice groups were infected by subcutaneous injection with 100 cercariae of Egyptian strain of S. mansoni. All mice groups were sacrificed 8 weeks post-infection. Results:Parasitological parameters such as worms count, eggs/gram tissues of liver and intestine were studied. Biochemical parameters such as the activities of liver function tests (AST, ALT, ALP and γGT) were measured. The cellular immune responses were assessed by studying the cytokine measurement. Hepatic histopathological criteria and the morphological changes of adult worms were studied through the scanning electron microscope. Conclusion: The present results revealed that the UV-irradiated adult worm extract have high efficacy than SWAP as immunizing antigens as shows in our data, which reported that irradiation associated vaccine antigen was shown to achieve a higher protection in mice.

Protection Against Schistosoma Mansoni In Mice By using UV-Irradiation In Comparing With Soluble Adult Worm Antigen (SWAP)

Background: The present study aimed to investigate the dynamics of immunoregulatory factors implicated in the murine model of soluble adult worm antigen preparation (SWAP)and UV-irradiated adult worm extract-induced granulomatous hypo-responsivness. Material and Methods: Native male mice were classified into four groups. The first negative control (non-infected) group, the second positive control (infected) group, the third SWAP-vaccinated group and the fourth UV-irradiated adult worm extract-vaccinated group. The last two groups were injected subcutaneous by 0.2 ml (contained 100 µg protein) of the two used vaccines SWAP or UV-irradiated adult worm extract respectively, four weeks prior to infection. The 2nd, 3rd and 4th mice groups were infected by subcutaneous injection with 100 cercariae of Egyptian strain of S. mansoni. All mice groups were sacrificed 8 weeks post-infection. Results:Parasitological parameters such as worms count, eggs/gram tissues of liver and intestine were studied. Biochemical parameters such as the activities of liver function tests (AST, ALT, ALP and γGT) were measured. The cellular immune responses were assessed by studying the cytokine measurement. Hepatic histopathological criteria and the morphological changes of adult worms were studied through the scanning electron microscope. Conclusion: The present results revealed that the UV-irradiated adult worm extract have high efficacy than SWAP as immunizing antigens as shows in our data, which reported that irradiation associated vaccine antigen was shown to achieve a higher protection in mice.

The Tumor Antigen Repertoire Identified in Tumor-Bearing Neu Transgenic Mice Predicts Human Tumor Antigens

FVB/N mice transgenic for nontransforming rat neu develop spontaneous breast cancers that are neu positive and estrogen receptor negative, mimicking premenopausal human breast cancer. These animals have been widely used as a model for immunobased therapies targeting HER-2/neu. In this study, we used serological analysis of recombinant cDNA expression libraries to characterize the antigenic repertoire of neu transgenic (neu-tg) mice and questioned the ability of this murine model to predict potential human tumor antigens. After screening 3 x 10(6) clones from 3 different cDNA libraries, 15 tumor antigens were identified, including cytokeratin 2-8, glutamyl-prolyl-tRNA synthetase, complement C3, galectin 8, and serine/threonine-rich protein kinase 1. Multiple proteins involved in the Rho/Rho-associated, coiled coil-containing protein kinase (Rock) signal transduction pathway were found to be immunogenic, including Rock1, Rho/Rac guanine nucleotide exchange factor 2, and schistosoma mansoni adult worm antigen preparation 70. All of the identified antigens are self-proteins that are expressed in normal tissues in addition to breast tumors and the majority of the antigens are intracellular proteins. More than half of the mouse tumor antigens have human homologues that have been reported previously as tumor antigens. Finally, the tumor-specific antibody immunity and marked immune cell infiltration that was observed in mice with spontaneous tumors were not observed in mice with transplanted tumors. Our results indicate that neu-tg mice bearing spontaneous tumors develop humoral immunity to their tumors similar to cancer patients and that tumor antigens identified in transgenic mouse may predict immunogenic human homologues.

Placental transfer of immunoglobulins in cattle infected with Schistosoma mattheei

Although the epitheliochorial placenta of ruminants does not allow passage of immunoglobulins from dam to foetus specific antibodies have been detected at birth in calves born to Schistosoma mattheei-infected cows. The present study determined the prevalence of calves born with specific antibodies for S. mattheei and the origin of these antibodies. For the determination of the prevalence, 100 calves born to infected mothers in an endemic area (Zambia) were examined, 24 were seropositive. To study the origin of these antibodies placentomes of 40 naturally S. mattheei-infected cows were examined for the presence of schistosome eggs and lesions which could explain foetal priming and/or leakage of maternal antibodies and/or antigen into the foetus. Tissue damage and schistosome eggs were observed on the maternal as well as the foetal side of the placentomes. In order to determine the specific nature of the antibody response, antibody profiles against soluble adult worm antigen preparation (SWAP) of S. mattheei were compared by Western blot between dams and their newborn calves (n = 8). The specific recognition profiles were identical for the seropositive calves and their dams on SWAP mattheei. Identical recognition profiles between dams and calves were also observed when sera were analysed on Escherichia coli, a pathogen of which the foetus should be free, and would indicate passive antibody transfer from the dam. In conclusion, the present study shows that S. mattheei could induce placentome lesions and that eggs can cross the placenta. Consequently, foeti can come into contact with S. mattheei antigens in utero, and might also contain maternal antibodies from leakage through placentome lesions. As such, the infection status of the mother could have far reaching effects on the immunological status of her offspring and modify their reaction upon infection.

The role of IL-10 and IgG1 in the protection and granulomatous response in Schistosomamansoni P24-immunized mice

Previous work by our laboratory identified a fraction of Schistosoma mansoni soluble adult worm antigenic preparation, designated PIII, able to elicit significant in vitro cell proliferation, and lower in vitro and in vivo granuloma formation. In the present work, we investigated some biological activities of P24, an antigenic component of PIII. Immunization of mice with this antigen induced a significant protection degree against challenge infection and significant decrease in the hepatic granuloma formation. Pre-incubation of spleen cells from P24-immunized mice with S. mansoni antigens induced a significant increase of interleukin (IL)-10 levels, but not interferon-γ, in the cell supernatants. In addition, mice immunized with different S. mansoni antigens and P24 displayed indistinguishable levels of IgG2a in response to anti- S. mansoni antigens, while IgG1 levels were significantly increased. Collectively, our results indicate that P24 might mediate protective anti-parasite immunity and downregulate granulomatous hypersensitivity to S. mansoni eggs in part by its ability to induce a higher production of IgG1 and IL-10.

Schistosoma mansoni in mice: the pattern of primary cercarial exposure determines whether a secondary infection post-chemotherapy elicits a T helper 1- or a T helper 2-associated immune response.

Reinfection with Schistosoma mansoni following chemotherapy often results in an ameliorated granulomatous reaction and hence a mild disease. This study examined some of the immunological mechanisms that could be associated with this residual protection. BALB/c mice were infected with either a single dose (group A) of 100 S. mansoni cercariae or with 10 doses of 10 cercariae each (group B) given at 3-day intervals. The mice were treated with praziquantel 8 weeks postinfection and, 2 weeks later, together with another group of naive mice (group C), they were infected with a single dose of 100 cercariae each. All the animals were killed 8 weeks later and schistosome egg antigen (SEA)- and soluble adult worm antigen preparation (SWAP)-induced cytokine recall responses in splenocytes, as well as serum immunoglobulin levels, were quantified and hepatic granuloma sizes measured. Group A animals had higher levels of SEA-induced interferon-gamma (IFN-gamma) but lower levels of interleukin (IL)-5 than groups B and C (P < 0.01). Group B animals had low SEA-induced IFN-gamma levels and elevated IL-5 levels, although these were lower than group C. SEA-induced IL-10 was low in both groups A and B as compared to group C (P < 0.01). SWAP was less effective as an inducer of splenocyte cytokine production than SEA but both SWAP-induced IFN-gamma and IL-5 were detected in groups A and C. SEA- and SWAP-specific immunoglobulin E (IgE) and immunoglobulin G (IgG) titres were not significantly different between the three groups. Granuloma diameters were larger in group C (mean 297 +/- 51.3 microm) as compared to groups A (174 +/- 49 microm, P < 0.01) and B (247.5 +/- 44 microm, P < 0.05). Taken together, these results demonstrate that granuloma size is reduced during a reinfection exposure compared with a primary infection. This reduction is associated with a T helper 1 response in mice exposed to a single large dose of cercariae in the primary infection and with a predominantly T helper 2 response in those infected with multiple small doses.

Parasite-specific antibody profile in human fascioliasis: application for immunodiagnosis of infection.

The antibody isotype response to an adult Fasciola worm antigen preparation (FWAP) was examined in sera from 60 Egyptians with parasitologically confirmed fascioliasis by an ELISA. The FWAP-specific IgG1 and IgG4 antibodies were found in 97-100% of the patients. The ratio of the mean absorbance values between infected patients and healthy controls was 9.7 and 29.7 for IgG1 and IgG4 antibodies, respectively. The IgM, IgA, IgG2, and IgG3 antibodies were less dominant. In contrast to IgG1 antibodies, which were often detected in sera from patients infected with Schistosoma, Echinococcus granulosus, Ascaris lumbricoides, Ancylostoma duodenale, or Hymenolepis nana, FWAP-specific IgG4 antibodies were detected exclusively in the sera of patients with fascioliasis. The data thus support the conclusion that an IgG4/ELISA with crude FWAP as antigen may be used for sensitive and accurate immunodiagnosis of human fascioliasis.

Familial resemblance in humoral immune response to defined and crude Schistosoma mansoni antigens in an endemic area in Brazil.

This study addressed whether the humoral immune response to crude and defined Schistosoma mansoni antigens aggregates within families. The sample included 155 siblings from 42 nuclear families in Brazil. Sera examined by ELISA for antibody isotypes reactive to defined schistosome antigens and crude schistosome antigens (soluble adult worm antigen preparation and soluble egg antigen) demonstrated that there was a difference in sibling-pair correlations between defined and crude S. mansoni antigens. In contrast to the finding with crude antigens, egg-positive sibling pairs showed significant familial resemblance for all IgG subclasses and IgE to adult-stage antigens Smp20.8 and Smp50. Only the IgE and IgG4 isotypes showed familial resemblance to the egg-stage antigen, Smp40. Egg-negative sibling pairs showed significant familial resemblance only for IgE and IgG4 to Smp40. That both the IgE and IgG4 response to defined S. mansoni antigens showed familial resemblance is interesting in light of the converging evidence for the role of IgE and IgG4 in human susceptibility and resistance to reinfection.

Schistosomiasis mansoni: immunoblot analysis to diagnose and differentiate recent and chronic infection.

One hundred seven patients classified into three different groups (11 with acute schistosomiasis, 58 with chronic schistosomiasis, and 38 children with high IgM-specific antibody titers against schistosome gut-associated antigens living in an endemic schistosomiasis area) were studied by immunoblotting for the presence of IgG, IgM, and IgA antibodies against Schistosoma mansoni soluble adult worm antigen preparation. We used sera from 15 individuals infected with various intestinal parasites, as well as sera from 19 uninfected individuals, as controls. An immunogenic fraction with a molecular weight of 31-32 kD (Sm31/32) was the most frequently recognized by the different antibody isotypes. In the group with acute disease, this fraction was recognized by IgG and IgM antibodies of all patients, and by 10 (90.9%) of 11 samples for IgA antibodies. Approximately 98% of the patients with chronic infections had IgG antibodies against Sm31/32, but only about 10% had IgM and IgA antibodies against this fraction. The IgG immunoblot profiles of the children from the endemic area were similar to those obtained for the group with acute schistosomiasis. This observation suggests recent infection of these children. Our data show that the Sm31/32 protein fraction is highly immunogenic and may be a useful serologic marker for diagnosing and differentiating between acute and chronic schistosomiasis infection.

Cytokine production in acute versus chronic human Schistosomiasis mansoni: the cross-regulatory role of interferon-gamma and interleukin-10 in the responses of peripheral blood mononuclear cells and splenocytes to parasite antigens.

The contribution of interleukin (IL)-10 and interferon (IFN)-gamma to the regulation of type 1 and type 2 cytokine responses was investigated in Brazilians with different clinical forms of schistosomiasis mansoni. Cells from members of a family with acute intestinal schistosomiasis responded to schistosomal soluble egg antigen (SEA) or soluble adult worm antigen preparation (SWAP) with greater amounts of IFN-gamma than did cells from several patients with chronic intestinal schistosomiasis; IL-10 levels were similar. Neutralization of IL-10 had no effect on the SEA-specific IFN-gamma response in patients with acute infection, whereas SWAP-induced IFN-gamma was increased in both groups. Anti-IL-10 also up-regulated SEA-specific IFN-gamma protein and mRNA responses in most splenocyte cultures from hepatosplenic schistosomiasis patients but had no effect on antigen-specific IL-4 or IL-5 production. Neutralization of IFN-gamma resulted in a comparable increase in SWAP-specific IL-10 and IL-5, while IL-4 was not affected. These studies demonstrate that early disease in schistosomiasis is associated with a significant IFN-gamma response and that IL-10 contributes to the suppression of that response during both early and chronic infection.

Nitric oxide and IL-10 production induced by PIII—A fraction of Schistosoma mansoni adult worm antigenic preparation—associated with downregulation of in vitro granuloma formation

Identification and characterization of Schistosoma mansoni antigens that can provide protective immunity, as well as an investigation of their role in host-parasite interaction, is crucial for understanding the complex immunoregulatory events that modulate granuloma formation in schistosomiasis. Previous work by our laboratory identified a fraction of S. mansoni soluble adult worm antigenic preparation (SWAP), named PIII, obtained by anionic chromatography on fast protein liquid chromatography (FPLC). This fraction was able to elicit significant in vitro cell proliferation and at the same time lower in vitro and in vivo granuloma formation when compared either to SWAP or to soluble egg antigens (SEA). In the present work, we investigate some biological activities of PIII, such as the stimulation of nitric oxide (NO) and cytokine production. Our data demonstrated that SEA, SWAP and specially PIII were able to induce a time-dependent increased NO production during in vitro granuloma reaction. Besides that, PIII evoked increased IL-10 production, but not IL-2 or IFNγ. Collectively, our results indicate the possibility that the modulation role of PIII on in vitro granuloma might be mediated in part by its ability to induce the higher production, initially of IL-10, and lately of NO.

Calorimetry: a highly sensitive technique for evaluating the effect of IL-2, IFN-γ and IL-10 on the response of peripheral blood mononuclear cells

We report the application of a calorimetric method for evaluating the metabolic response of peripheral blood mononuclear cells (PBMC) either from S. mansoni infected patients or from normal subjects under stimulation with antigen or with cytokines. Our results demonstrate that the antigenic stimulation of the PBMC with soluble adult worm antigenic preparation (SWAP) was able to discriminate PBMC from infected and non-infected donors, based on calorimetric cell reactivity. The total heat release in the stimulation of PBMC with interleukin 2 (IL-2) or gamma interferon (IFN-γ) was greater for PBMC from infected patients than normal subjects. The activation of the release heat in PBMC induced by IFN-γ, IL-2 or antigen (SWAP) was inhibited by pretreatment of PBMC with interleukin 10 (IL-10). However, this inhibition mediated by IL-10 could be promptly reversed by pretreatment of IL-10 with its respective anti-IL-10 monoclonal antibody. Our present results extend the use of the calorimetric assay and suggest its application for studying cellular reactivity in basic and applied immunological systems.