Biomedical subjects

A M Deelder

Publications and source records attributed to A M Deelder.

At least 37 records · Page 2Linked to original sources

PubMed2000

Specificity of circulating antigen detection for schistosomiasis mansoni in Senegal and Burundi.

The specificity of schistosome circulating antigen detection was determined in negative individuals from two S. mansoni- endemic countries, Senegal and Burundi, and compared with results from Dutch control individuals. A nearly absolute specificity was achieved for circulating anodic antigen (CAA) detection in serum, irrespective of the target population or sample pretreatment method. Circulating cathodic antigen (CCA) detection in serum and urine resulted in a lower specificity than serum CAA detection. Apparent large differences in specificity of CCA detection between countries were mainly due to pretreatment methods. Apparently, the alkaline/heating pretreatment method is not as effective as trichloroacetic acid (TCA)-pretreatment in removing (certain) interfering components, which may vary between populations. In view of the development of the urine CCA assay into a noninvasive screening test, a slightly lower specificity may still be acceptable. For precise epidemiological analyses the highly specific serum CAA assay remains the method of choice.

Adolescent↗

PubMed2000

Study of the distribution pattern of Schistosoma haematobium egg antigens recognised by six different monoclonal antibodies in the parasite and the host.

Recently a new panel of monoclonal antibodies was developed against soluble egg antigens in the hatching fluid of Schistosoma mansoni. These antibodies have been used to develop an improved ELISA for the detection of circulating soluble egg antigens in serum and urine that would have a higher sensitivity in the immunodiagnosis of S. mansoni infections. Although these antibodies showed no improvement in the immunodiagnosis of S. mansoni infections compared with egg antigen-based ELISAs already described (Nourel Din et al., 1994a), they may have a potential role in the identification of S. haematobium infections. This study has looked into the immunolocalisation of S. haematobium egg antigens in both the parasite and the host as recognised by four newly developed monoclonal antibodies (290-2D9-A, 290-2E6-A, 290-2H12-A and 290-4A8-A) and two already described antibodies (114-5B1-A and 114-4D12-A). The antibodies 114-5B1-A and 114-4D12-A appeared to have in S. haematobium eggs a similar staining pattern when compared to S. mansoni eggs. The antibodies prepared against the hatching fluid showed a characteristic signal, especially 290-2E6-A. These antibodies recognised a component originating from the lateral glands of the miracidium. In the host a similar immunohistochemical tissue localisation pattern (mainly phagocytising reticulo-endothelial cells) was seen as previously described for S. mansoni infected hamsters.

Animals↗

PubMed2000

Various stages of schistosoma express Lewis(x), LacdiNAc, GalNAcbeta1-4 (Fucalpha1-3)GlcNAc and GalNAcbeta1-4(Fucalpha1-2Fucalpha1-3)GlcNAc carbohydrate epitopes: detection with monoclonal antibodies that are characterized by enzymatically synthesized neoglycoproteins.

We report here that fucosylated epitopes such as Lewis(x), LacdiNAc, fucosylated LacdiNAc (LDN-F) and GalNAcbeta1-4(Fucalpha1-2Fucalpha1-3)GlcNAc (LDN-DF) are expressed by schistosomes throughout their life cycle. These four epitopes were enzymatically synthesized and coupled to bovine serum albumin to yield neoglycoproteins. Subsequently these neoglycoproteins were used to probe a panel of 188 monoclonal antibodies obtained from infected or immunized mice, in ELISA and surface plasmon resonance analysis. Of these antibodies, 25 recognized one of the fucosylated structures synthesized, indicating that these structures are immunogenic during infection. The MAbs identified could be subdivided in four different groups based on the recognition of either the Lewis(x)-, the LacdiNAc-, the LDN-DF-, or both the LDN-F- and LDN-DF epitope. These monoclonal antibodies were then used to investigate the localization of the fucosylated epitopes in various stages of Schistosoma mansoni using indirect immunofluorescence. Lewis(x)epitopes were mainly found in the gut and on the tegument of adult worms, on egg shells, and on the oral sucker of cercariae. The LacdiNAc epitope was expressed on the tegument of adult worms, on miracidia, and on the oral sucker of cercariae. In contrast, LDN-DF epitopes were mainly present in the excretory system of adult worms, on miracidia and on whole cercariae. These also stained positive with the LDN-F/LDN-DF epitope antibodies, while whole parenchyma reacted characteristically only with the latter antibodies. The identification of different carbohydrate structures in various stages of schistosomes may lead to a better understanding of the function of glycans in the immune response during infection.

Animals↗

PubMed1999

Development of antibody isotype responses to Schistosoma mansoni in an immunologically naive immigrant population: influence of infection duration, infection intensity, and host age.

We have identified the influence of host and parasite factors that give rise to characteristic antibody isotype profiles with age seen in human populations living in different areas of schistosomiasis endemicity. This is important in the immunobiology of this disease. It is also of interest in the context of human responses to chronic antigen stimulation, vaccines, allergens, and other pathogens. In populations exposed to endemic schistosomiasis, factors such as intensity and duration of infection are age dependent. They therefore confound the influence of host age on antiparasite responses. Here, we resolved these confounding factors by comparing the developing antibody responses of an immunologically naive immigrant population as they acquired the infection for the first time with those of chronically infected resident inhabitants of the same region of Schistosoma mansoni endemicity in Kenya. Recent arrival in the area strongly favored immunoglobulin G3 (IgG3) responses against the parasite. The antibody isotype responses associated with human susceptibility to reinfection after chemotherapy were elevated in those suffering high intensities of infection (IgG4 responses against worm and egg antigens) or were characteristic responses of young children irrespective of the intensity or duration of infection (IgG2 responses against egg antigen). IgE responses against the adult worm, a response associated with resistance to reinfection after chemotherapy, increased with the ages of infected individuals and were also favored in those currently suffering higher intensities of infection.

Adult↗

PubMed1999

Application of immunodiagnostic assays: detection of antibodies and circulating antigens in human schistosomiasis and correlation with clinical findings.

In an initial cross-sectional survey, serum, urine, and stool samples were collected from 370 participants representing about 10% of the population (n = 4,438) in Behbeet village, 50 km south of Cairo, Egypt, an area well known to be endemic solely for Schistosoma haematobium. Diagnosis was approached in two parallel ways. The first approach, which simulated actual conditions in many endemic areas in Egypt, was based on physical examination and urine and stool microscopic analysis. The second approach was based on two advanced immunodiagnostic assay systems. One system detected antibodies to species-specific microsomal antigens, the other detected circulating schistosomal antigens. Microsomal antigens from S. haematobium and S. mansoni were used to detect antibodies in the Falcon assay screening test (FAST)-ELISA and the enzyme-linked immunoelectrotransfer blot (EITB). Circulating anodic antigen (CAA) and circulating cathodic antigen (CCA) were quantified in serum and urine samples in a sandwich ELISA using monoclonal antibodies. Parasitologically, the prevalence of S. haematobium was 7.01% in females and 25.82% in males, giving an overall prevalence of 15.8%. The combination of urine CCA and serum CAA for detecting circulating antigens and the combination of the S. haematobium adult worm microsomal antigens (HAMA) FAST-ELISA and the HAMA EITB for detecting antibodies significantly improved the sensitivity of detecting S. haematobium circulating antigens and antibodies. Also, including a medical examination as an integral part of field studies and correlating immunodiagnostic results with other clinical and investigational data allowed us to calculate an accurate estimation of S. haematobium prevalence in this area of low endemicity.

Adolescent↗

PubMed1999

Urine circulating soluble egg antigen in relation to egg counts, hematuria, and urinary tract pathology before and after treatment in children infected with Schistosoma haematobium in Kenya.

A cohort of 117 school children infected with Schistosoma haematobium was followed-up after therapy with praziquantel (0, 2, 4, 6, 12, and 18 months) and various infection and morbidity parameters (egg counts, hematuria, soluble egg antigen [SEA] in urine, and ultrasonography-detectable pathology) were quantified. At the onset of the study, 97% of the children were positive for S. haematobium with a geometric mean egg count of 45.7 eggs/10 ml of urine. Eighty-one percent of the children were positive for SEA in urine with a geometric mean SEA concentration of 218.8 ng/ml of urine. Ninety-two percent and 56% of the children were microhematuria positive and macrohematuria positive, respectively. Two months after treatment, all infection and morbidity indicators had significantly decreased. Reinfection after treatment as determined by detection of eggs in urine was observed by four months post-treatment while the other parameters remained low. The clearance of SEA was slower than that of egg counts while pathology resolved at an even slower pace. Levels of SEA and egg output showed similar correlations with ultrasound detectable pathology; these correlations were better than the correlation between hematuria and pathology.

Adolescent↗

PubMed1999

[Purification and specific detection of two major Schistosoma gut-associated circulating antigens, CAA and CCA].

AIM: To investigate the specificity differences of the 2 major gut-associated diagnostic circulating molecules in Schistosoma japonicum infection, CAA and CCA, and to obtain sufficient purified material for setting up a standard series in quantitative determinations. METHODS: Isolation and purification of the two worm fractions from a trichloroacetic acid (TCA) soluble preparation of S. japonicum adult worm antigen (AWAj-TCA) by Mono-Q anion exchange chromatography were performed and the specific reactivity of the eluted fractions by antigen-capture ELISA (specific for CAA or CCA) with reference to affinity purified preparations of S. mansoni CAA and CCA was analysed. RESULTS: By using an ionic strength gradient, CCA was eluted in two major peaks, an unbound fraction CCA-1, and a major bound fraction CCA-2. Two additional minor peaks, CCA-3 and CCA-4, were eluted at higher ionic strengths. CAA was only detected in a bound fraction, partly overlapping with CCA-3. In the CCA-1 and CCA-2 fractions reactivity was only found in the antigen-capture ELISA using anti-CCA McAbs both for capture and detection. The CAA fraction was predominantly found to be positive in the antigen-capture ELISA using anti-CAA McAbs both for capture and detection. However, when using combinations of anti-CCA and anti-CAA McAbs for capture and detection by ELISA this fraction showed some reactivity. CONCLUSION: Two CCA fractions contain molecules which bear at least two CCA-epitopes; while the CAA fraction contains molecules which contain at least two CAA-epitopes, and one CCA-epitope.

Animals↗

PubMed1998

Adhesion molecules in intestinal Schistosoma mansoni infection.

Adhesion molecules constitute essential elements in inflammation, mediating various cellular interactions. We investigated the expression of adhesion molecules mediating cell-cell [intercellular adhesion molecule-1 (ICAM-1) and lymphocyte function-associated antigen-1 (LFA-1)] and cell-matrix interactions [very late antigen-4 (VLA-4), VLA-6, and syndecan-1] in intestinal granulomas of mice infected with the parasite Schistosoma mansoni. Up-regulation of ICAM-1, LFA-1, and VLA-4 was seen in ileal and colonic granulomas, at both the acute (8 weeks postinfection) and the chronic stage (13-16 weeks postinfection). Up-regulation of VLA-6 was absent in all intestinal granulomas. Syndecan-1 immunoreactive (antigen-driven) B-lymphocytes were seen in the proximity of egg-antigen-laden macrophages in the inner part of ileal and colonic granulomas, although B-cells are considered to be absent in ileal granulomas. Estimation of intestinal granuloma volumes demonstrated the lack of down-modulation observed in ileal granulomas. From our results we infer that adhesion molecules constitute important elements in schistosomal intestinal granuloma formation. Organ-related differences between hepatic and intestinal granulomas exist (e.g., granuloma volume), but these differences are not morphologically reflected in a differential expression of the adhesion molecules ICAM-1, LFA-1, and VLA-4. Syndecan-1 immunoreactive B-lymphocytes also appear to be involved in ileal granuloma formation.

Animals↗

PubMed1998

The effect of anti-IL-10 on proliferation and cytokine production in human schistosomiasis: fresh versus cryopreserved cells.

In this study we neutralized endogenous IL-10 in PBMC from individuals chronically infected with Schistosoma haematobium by using anti-IL-10 MoAbs, and examined the effect and adult worm antigen (AWA)-specific responses in both fresh or cryopreserved cells. Anti-IL-10 alone increased background proliferation of PBMC, but did not augment the AWA-specific responses in either fresh or frozen cells. In freshly isolated PBMC, IFN-gamma production in response to AWA was enhanced significantly in the presence of anti-IL-10. In cryopreserved cells, the augmentation of IFN-gamma in the presence of anti-IL-10 was four-fold less than in the freshly-isolated cells. Neutralization of IL-10 had no effect on IL-4 production. These data show that IL-10 plays a role in specifically down-regulating Th1-but not Th2-type responses and, in contrast to previous reports, anti-IL-10 does not augment proliferation to parasite antigen in chronic schistosomiasis.

Animals↗

PubMed1998

Antigen-specific proliferation and interferon-gamma and interleukin-5 production are down-regulated during Schistosoma haematobium infection.

Antigen-specific cellular immune responses were examined in persons previously infected with Schistosoma haematobium and who were, 2 years after chemotherapy, either free from infection (n = 17) or reinfected (n = 20). Proliferation to adult worm antigen (AWA) was significantly higher in uninfected than in reinfected subjects (P = .02), whereas responses to soluble egg antigen (SEA) remained low in both groups. Interleukin (IL)-5 production in uninfected persons in response to AWA and SEA was higher than in infected subjects (P = .05 and P < .001, respectively), while IL-4 and IL-13 release was similar in the 2 groups. Levels of interferon-gamma to AWA and to SEA were inversely correlated with egg output (P = .03 and P = .02, respectively). These data indicate that the presence of schistosome infection leads to T cell proliferative hyporesponsiveness and that both typical Th1 and Th2 cytokines can be down-regulated by active infection.

Adolescent↗

PubMed1998

Human IgE, IgG subclass, and IgM responses to worm and egg antigens in schistosomiasis haematobium: a 12-month study of reinfection in Cameroonian children.

Levels of IgE, IgM, and IgG subclasses against Schistosoma haematobium adult worm antigen (AWA) and soluble egg antigen (SEA) in a cohort of 148 S. haematobium-infected schoolchildren were determined before and up to 12 months after chemotherapy. Infection intensities were determined as concentrations of circulating anodic antigen (CAA) in serum. One month posttreatment, the antibody levels of all isotypes against AWA were increased, but 1 year after treatment they returned to pretreatment levels. CAA concentrations were positively associated with levels of IgG4 against AWA and SEA but not with levels of IgE. Age correlated negatively with CAA concentrations and positively with levels of IgE to AWA. The balance of anti-AWA IgG4 and IgE was significantly correlated to the CAA concentration, in particular in the older age group (11-13 years). This may suggest that protective immune mechanisms in S. haematobium infections become effective around the age of 12 years.

Adolescent↗

PubMed1998

Advances in schistosomiasis research.

This review reports on recent developments in the areas of diagnosis, chemotherapy and pathology, where female genital schistosomiasis was recently identified as a common cause of morbidity in Schistosoma haematobium-endemic areas. As a profound understanding of processes underlying resistance and susceptibility to infection as well as pathology is central to schistosomiasis control, detailed immunological studies of immunity and granuloma formation dominate the discussion. Studies on host genetics, on T-helper cell type 1 or 2 cytokines as well as on cell types influencing immunity and granuloma formation have provided precise information that could be of relevance not only to helminth but also to microbial infections involving granulomatous reactions.

Journal Article↗

PubMed1998

Age-related worm load and worm fecundity patterns in human populations, as indicated by schistosome circulating antigens.

Recently, our group determined the relationship between serum CAA levels and fecal egg counts in two foci with very intense Schistosoma mansoni transmission: Maniema (Zaire), an area endemic for S. mansoni since several decades, and Ndombo (Senegal), where transmission has only been established since a few years. The objective was to study and compare age-related worm load and worm fecundity patterns in these two different endemic settings. Here, we will summarize the most important findings and conclusions of this study.

Africa↗

PubMed1998

Use of monoclonal antibodies prepared against Schistosoma mansoni hatching fluid antigens for demonstration of Schistosoma haematobium circulating egg antigens in urine.

A panel of 17 monoclonal antibodies (MAbs) against Schistosoma soluble egg antigens (SEAs) was produced from BALB/c mice immunized with antigens secreted/excreted by Schistosoma mansoni eggs. In this study, we demonstrate that 16 MAbs were reactive with S. haematobium SEA in addition to S. mansoni SEA. The MAbs were tested as potential immunodiagnostic reagents in a homologous sandwich ELISA format to detect circulating soluble egg antigens (CSEAs) in serum and urine samples of S. mansoni- or S. haematobium-infected individuals. When samples of S. mansoni-infected individuals were tested, none of these MAbs performed as good as the previously described S. mansoni-specific 114-5B1-A and 114-4D12-A MAbs. However, 11 MAbs (of the IgM isotype) detected CSEA in urine samples of S. haematobium-infected individuals. Three MAbs, 290-2E6-A, 291-3D5-A, and 291-5D5-A, were selected for a pilot study with 47 urine samples of S. haematobium-infected individuals from Kenya. The CSEA levels detected with each of these ELISAs showed a significant correlation with urinary egg counts (Spearman rho > 0.37, P < 0.01) and with each other (Spearman rho > 0.74, P < 0.001). Based on the 92% specificity and 90% sensitivity of the assay, the ELISA using MAb 290-2E6-A was found to be the most promising assay for immunodiagnosis of S. haematobium infections.

Animals↗

PubMed1998

Day-to-day fluctuation of schistosome circulating antigen levels in serum and urine of humans infected with Schistosoma mansoni in Burundi.

Day-to-day fluctuations of both circulating anodic antigen (CAA) and circulating cathodic antigen (CCA) in serum and urine were examined simultaneously in a group of Schistosoma mansoni-infected individuals from Burundi and compared with each other and with fecal egg count fluctuations. Significant correlations were found between fecal egg counts and circulating antigens (CAA and CCA) and between circulating antigen levels in serum and urine samples. The cumulative percentage of positive results after three samplings was highest for urine CCA detection, followed by fecal egg counts, serum CCA, serum CAA, and urine CAA detection, respectively. It was demonstrated that circulating antigen levels in both serum and urine showed less fluctuation than fecal egg counts, except for urine CAA levels. The serum CAA detection assay in particular, although less sensitive in this low endemic area in Burundi, gave very constant measurements over a period of one week. Our results indicate that detection of circulating antigens in a single serum or urine sample provides a quantitatively more stable diagnosis of S. mansoni infection than fecal egg counts based on a single stool examination.

Adolescent↗

PubMed1998

Detection and quantification of soluble egg antigen in urine of Schistosoma haematobium-infected children from Kenya.

While research on alternative diagnostic and morbidity markers for infection with Schistosoma haematobium has been going on for a long time, egg counts continue to be used as the gold standard, and infection intensity is thought to reflect the severity of the disease. However, this relationship is not always clear and fluctuation in egg output makes it difficult to classify prevalence correctly. The use of circulating adult worm antigen detection as an alternative diagnostic technique has been applied with varying success. However, this is a measure of worm burden and does not reflect the tissue egg load(s). In the present study we have used an assay that detects soluble egg antigen (SEA) in urine of S. haematobium-infected children, and we have evaluated the applicability of the assay as a diagnostic and morbidity indicator. To evaluate this assay, we have studied a group of 470 children from two schools (Tsunguni and Kibaokiche) in the Coast province of Kenya; 84.8% and 77% were egg-positive while the percentage positive as determined by the SEA-ELISA were 78.8% and 76.2% in Tsunguni and Kibaokiche, respectively. In both schools, SEA levels in urine of S. haematobium-infected children significantly correlated with egg counts (Pearson's r=0.73, P < 0.0001) and with hematuria (Spearman's r=0.65, P < 0.0001). In addition, urinary tract pathology as determined by ultrasound significantly correlated with the SEA levels in urine (Spearman's r=0.3, P < 0.001). The SEA-ELISA compared well with microhematuria within egg count classes and with egg counts within hematuria classes.

Adolescent↗

PubMed1997

Sensitivity of microscopy versus enzyme immunoassay in the laboratory diagnosis of giardiasis.

The substitution of enzyme immunoassay (EIA) techniques for microscopy as a screening tool for Giardia lamblia infection was assessed. Paired stool samples obtained within a ten-day period from 366 patients with persistent diarrhea were examined by microscopy. In addition, two commercially available Giardia lamblia-specific EIAs were performed. Compared with microscopy, EIA for copro-antigen detection was more sensitive, based on examination of either one or two stool samples. Repeated examinations increased the number of cases detected, more so for microscopy than EIA. The negative predictive values of the two EIAs performed on the first stool sample were 98.7% and 97.8%. The results show that EIA for detection of copro-antigens in a single stool sample may be almost as sensitive for identifying Giardia infection as repeated microscopy on two sequential stool samples.

Adolescent↗

PubMed1997

Enhanced detection of Schistosoma circulating antigens by testing 1 ml urine samples using immunomagnetic beads.

The potential advantage of increasing the sample volume for enhanced detection of two Schistosoma circulating antigens in urine has been evaluated. Two different, monoclonal antibody based, immunomagnetic beads assays detecting circulating anodic antigen (CAA) and circulating soluble egg antigen (CSEA), respectively, were investigated. By increasing the sample volume to 1.0 ml, the lower detection limit of antigen dilution series was improved six eight-fold in both immunomagnetic beads assays. Indeed, an improvement could be shown when testing the urine of Schistosoma-infected individuals with these magnetic beads assays. To compare the sensitivity of the immunomagnetic beads method with the standard ELISA, urine was experimentally adjusted to mirror low natural infection intensities. CSEA-positive urine samples were adjusted to levels below the detection limit of the corresponding ELISA and indeed were still found positive by the CSEA immunomagnetic beads assay. These results demonstrate that by using immunomagnetic beads, an assay method which can utilise larger sample volumes than an ELISA, enhanced detection can be achieved. This might be particularly valuable in areas with low prevalence of schistosomiasis or for the assessment of cure after chemotherapy.

Animals↗