Evaluation of the prophylactic and therapeutic efficacies of mucus and tissue nucleoproteins extracted from Biomphalaria alexandrina snails on schistosomiasis mansoni.

Schistosomiasis is a neglected tropical disease with considerable morbidity. The lone effective drug, praziquantel (PZQ), is showing emergence of drug resistance hence, searching for new supportive treatment is crucial. This study aimed to evaluate the efficacy of mucus and nucleoproteins (NPs) extracted from Biomphalaria alexandrina (B. alexandrina) snails on miracidia, cercariae and Schistosoma mansoni (S. mansoni) adults in vitro and assess their experimental in vivo effect through parasitological, histopathological, and biochemical parameters. The in vivo study included 90 male Swiss albino mice. Mice were grouped into 9 groups; G1-G5 were infected and treated with; GI: PZQ, GII: mucus, GIII: combined PZQ and mucus, GIV: NPs, GV: combined PZQ and NPs. Control groups; C1: Non infected non treated (negative control), C2: Infected non treated (positive control), C3: Non infected mucus treated and C4: Non infected NPs treated. The in vitro study proved that the mucus had a better lethal effect on cercariae than miracidia, while NPs had better lethal effect on miracidia. The mucus lethal effect on adults surpassed the NPs as 100% and 60%, respectively. The in vivo study proved that the combined NPs or mucus with PZQ added to the effect of individual PZQ resulting in 100% total worm burden (TWB) reduction. As regard oxidative stress markers, the lowest level of nitric oxide (NO) was shown with combined PZQ and NPs. While, the highest glutathione (GSH) level was produced by individual PZQ. The study concluded that mucus and NPs of B. alexandrina had cercaricidal, miracidicidal and anti-schistosomal effect in vitro and that their combination could be considered a contribution to PZQ potentiality in vivo.

The potential use of 29 kDa protein as a marker of pathogenicity and diagnosis of symptomatic infections with Blastocystis hominis.

The present study was performed to characterize the protein profiles of Blastocystis hominis isolates from symptomatic and asymptomatic individuals by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting using sera from symptomatic and asymptomatic patients. The presence of immunogenic bands associated with pathogenicity or of diagnostic potentials was also evaluated. The study comprised 80 individuals classified into four groups, 20 each: symptomatic blastocystosis (G1), asymptomatic blastocystosis (G2), other parasitic infections (G3), and healthy control subjects (G4). SDS-PAGE analysis of individual antigens form symptomatic and asymptomatic B. hominis isolates revealed similar and distinctive antigenic bands with significant differences in two high (123.5 and 112.3 kDa) and few low molecular weight bands (48.5, 38, 42.3, and 35.5 kDa). Immunoblotting was performed using symptomatic and asymptomatic antigen pools with sera of the four studied groups. It was found that anti-B. hominis IgG reacted with nine protein bands ranging from 100 to 18 kDa of the symptomatic antigen pool. There was a significant difference between G1 and G2 in the recognition of 64, 56, 38, and 29 kDa antigen bands. Also, anti-B. hominis IgG reacted with five protein bands ranging from 56 to 12 kDa of asymptomatic antigen pool. There was a significant difference between G1 and G2 in the recognition of 29 kDa antigen band. These findings suggest the potential use of the 29-kDa antigen as marker of pathogenicity and implicate its use in the diagnosis and differentiation between symptomatic and asymptomatic blastocystosis.

Ancylostoma duodenale infection: a study of serum immunoglobulin G4 response to the excretory secretory antigen of adult worm.

The serum anti-Ancylostoma duodenale immunoglobulin (Ig) G4 antibody response to fraction III of the partially purified excretory secretory antigen of adult worm (Ad III ESA) was studied. The work included 60 patients with A. duodenale infection (GI), 40 patients with other parasitic infections (GII) and 30 apparently healthy parasite-free controls (GIII). Level of serum specific IgG4 was measured by indirect enzyme linked immunosorbent assay and compared with serum specific IgG, IgG 1, 2 & 3 subclass antibodies. Patients of GI had gastro-intestinal manifestations and symptoms suggestive of anaemia, and by investigations they had anaemia in 31.7% & eosinophilia in 100%. Measuring the intensity of A. duodenale infection, quantified as fecal egg counts, in patients of GI revealed that 60%, 30% & 10% had light, moderate, and heavy infections, respectively. The serum anti-Ad III ESA IgG & IgG 1-4 subclass antibodies were significantly elevated (P < 0.001) in patients of GI compared with GIII. Serum specific IgG4 was expressed in 100% of patients of GI at a significantly highly elevated level than IgGI (P < 0.01), IgG2 & IgG3 (P < 0.001). Specific IgG1 was expressed in 88.3% of patients of GI at a significantly elevated level (P < 0.001) than IgG2 & IgG3 which were expressed in 31.7% & 38.3%, respectively and elevated to a moderate extent. Serum specific IgG4 showed a 1.0, 1.1, 3.1 & 2.6-fold increase in detection rate of positive cases than IgG, IgG1, IgG2 & IgG3, respectively. The highest ability to differentiate between infected and healthy subjects was by serum specific IgG4 recording a discrimination coefficient of 9.4, while IgG, IgG1, IgG2 & IgG3 recorded 5.2, 6.3, 3.2 & 3.4, respectively. Serum specific IgG4 showed a significant positive correlation (r = 0.41, P < 0.001) with the intensity of A. duodenale infection, as was demonstrated by IgG & IgG3 (P < 0.01 & P < 0.05, respectively). Detection of serum anti-Ad III ESA IgG4 antibody recorded a 100% sensitivity that was significantly higher (P < 0.001) than IgG1, IgG2 & IgG3, but insignificantly different (P > 0.05) from IgG. Finally, serum specific IgG4 recorded a 100% specificity that was significantly higher than IgG, IgG2, IgG3 (P < 0.01) & IgG1 (P < 0.05). They showed cross-reactions with ascariasis, lymphatic filariasis and strongyloidiasis. The results are discussed.

Immunopathogenic role of IgG antibody and RANTES in house dust mite-induced chronic bronchitis.

Based on immunological and clinical examinations, 21 patients were diagnosed as having house dust mite (HDM)-induced chronic bronchitis and classified into three groups according to the clinical presentation of the disease: stable bronchitis, exacerbated bronchitis and asthma on top of bronchitis. Using ELISA, the levels of serum anti-Dermatophagoides farinae and anti-D. pteronyssinus IgG antibodies and plasma RANTES (regulated upon activation, normal T-cell-expressed and secreted; a chemokine with attractive and activator role for eosinophils) were measured in correlation to serum eosinophil cationic protein (ECP, a marker of eosinophil activation and degranulation measured by chemiluminescent immunometric technique). Using immunoblotting, IgG binding components of D. farinae and D. pteronyssinus were determined providing a clue for diagnosis of HDM-induced chronic bronchitis. Significant higher levels of anti-D. farinae and anti-D. pteronyssinus IgG antibodies and RANTES were found in asthmatic group followed by exacerbated chronic bronchitis in comparison to stable bronchitis and control groups. ECP level correlated significantly with IgG and RANTES levels in exacerbated bronchitis and asthmatic groups. The results provided evidence that over expression of IgG and RANTES plays a crucial role, as mediator in immunopathogenesis of HDM-induced chronic bronchitis and as marker of the immunological changes likely responsible for progression of bronchitis to asthma in HDM-sensitive patients yet, RANTES seemed to be an early indicator. Definition of the immunopathogenic role of IgG and RANTES in HDM-induced bronchitis should enable the manipulation of the critical immune response in the hope of establishing new therapies. D. farinae and D. pteronyssinu antigenic bands at > 205 and 205 KDa, respectively, considered together showed 71.4% sensitivity in diagnosis of HDM induced chronic bronchitis and 100% specificity by immuno-blotting.

Alkaline phosphatase from Echinococcus granulosus metacestodes for immunodiagnosis of human cystic echinococcosis.

The work examined the use of Echinococcus granulosus alkaline phosphatase (EgAP) (extracted from hydatid cyst membranes) as an antigen for immunodiagnosis of human cystic echinococcosis (CE). It was assessed by enzyme-linked immunosorbent assay (ELISA) and Western immunoblotting (IB) for detection of serum anti-EgAP immunoglobulin (Ig)G antibody and was compared with hydatid cyst fluid (HCF). The EgAP and HCF were of sheep liver cysts origin. Sera from 30 patients with surgically confirmed CE (G. I), 30 patients with other parasitic infections (G. II), and 20 healthy controls (G. II) were examined. The mean optical density of each of anti-EgAP IgG and anti-HCF IgG antibodies in G. I was significantly higher (P < 0.01) than in each of G. II and III. The use of EgAP in ELISA showed 100% sensitivity and specificity recording significantly higher sensitivity (P < 0.05) and specificity (P < 0.01) than when using HCF in ELISA which showed 86.7% sensitivity and 84% specificity. SDS-PAGE resolution, under reducing conditions, of EgAP revealed a molecular weight of 56 KDa, while that of HCF revealed a number of antigenic bands ranged from 12 to 130 KDa. IB analysis showed that sera from CE patients recognized the EgAP 56 KDa and also one or more of HCF antigenic bands of molecular weights at 116, 63, 44, 39, 24, 20, 16 and 12 KDa. The use of EgAP in IB showed 100% sensitivity and specificity recording an insignificant difference (P > 0.05) in sensitivity and a significantly (P < 0.05) higher specificity than when using HCF in IB which showed 100% sensitivity and 90% specificity. Cross reactivity with HCF in ELISA and IB was seen with schistosomiasis mansoni, fascioliasis, hymenolepiasis nana and ascariasis. Using EgAP, there was an insignificant difference (P > 0.05) in each of the sensitivity and specificity between ELISA and IB. Using HCF, there was a significantly (P < 0.05) higher sensitivity and an insignificantly (P > 0.05) higher specificity by IB than ELISA. The implications of these results are discussed.

Evaluation of schistosomicidal activity of myrrh extract: parasitological and histological study.

Ninety mice were divided into three main groups: G. I (non-infected control), G. II (infected non treated control) and G. III (infected treated), which was further subdivided into 4 subgroups: IIIA, IIIB, IIIC & IIID, where the drug was administered in a dose of 500 mg/kg body weight for five days before infection for subgroup IIIA, on the 1st day post infection (PI) for subgroup IIIB, 21 days PI for subgroup IIIC and 45 days post infection for subgroup IIID. All animals were sacrificed 80 days after the start of the experiment. Anti-schistosomal activity was assessed parasitologically by estimating the percentage reducetion of worm burden, egg count/gm tissues (liver & intestine), and the oogram pattern; histologically & histopathologically by examination of jejunum using different stains. The percentage reduction of worm burden was 30.35%, 64.54%, 76.92% and 98.46% respectively when compared to the infected non-treated control G. The maximum effect of the drug was observed in subgroups IIIC & IIID. Also, there was marked reduction in the egg count in tissues (liver & intestine). In addition the oogram pattern showed that myrrh had high antischistosomal activity. The histologically and histopathologically infected untreated (GII), when compared to non-infected non-treated control G. showed apparent shortening and flattening of the jejunal villi with focal loss of the epithelial covering. Loss of PAS positive brush border of many enterocytes with goblet cells hyperplasia was seen. Bilharzial granulomas were frequently encountered in the submucosa and the musculosa with numerous eosinophils content. In subgroups IIIA & IIIB, there was mild amelioration of the mucosal structural abnormalities. The granulomas were less frequently seen with decrease of their eosinophils. In subgroups IIIC & IIID there was restoration of the jejunal mucosal continuity, marked decrease in the granulomas and paucity of eosinophils. The present data proved that myrrh has a valuable schistosomicidal effect against different maturation stages of S. mansoni. The chemotherapeutic effect was more evident when the drug was given to the infected mice on the 21st as well as on the 45th day PI. The drug proved a promising chemoprophylactic agent when used five days before exposure to infection.