ß-glucan mitigates ovalbumin-induced airway inflammation by preventing oxidative stress and CD8+ T cell infiltration.

BACKGROUND: Bronchial asthma is a severe respiratory condition characterized by airway inflammation, remodeling, and oxidative stress. ß-Glucan (BG) is a polysaccharide found in fungal cell walls with powerful immunomodulatory properties. This study examined and clarified the mechanisms behind BG's ameliorativeactivitiesin an allergic asthma animal model. METHOD: BG was extracted from Chaga mushroom and characterized using FT-IR, UV-visible, zeta potential, and 1H NMR analysis. The mice were divided into five groups, including control, untreated asthmatic, dexamethasone (Dexa)-treated (1 mg/kg), and BG (30 and 100 mg/kg)-treated groups. RESULTS: BG treatment reduced nasal scratching behavior, airway-infiltrating inflammatory cells, and serum levels of IgE significantly. Additionally, BG attenuated oxidative stress biomarkers by lowering malonaldehyde (MDA) concentrations and increasing the levels of reduced glutathione (GSH), glutathione peroxidase (GPx), and catalase (CAT). Immunohistochemical and flow cytometric analyses have confirmed the suppressive effect of BG on the percentage of airway-infiltrating cytotoxic CD8+ T cells. CONCLUSION: The findings revealed the role of CD8+ T cells in the pathogenesis of asthma and the role of BG as a potential therapeutic agent for asthma management through the suppression of airway inflammation and oxidative stress.

Echinochrome exhibits anti-asthmatic activity through the suppression of airway inflammation, oxidative stress, and histopathological alterations in ovalbumin-induced asthma in BALB/c mice.

Asthma is a chronic pulmonary disease with marked infiltrating inflammatory cells and reduced respiratory performance. Echinochrome (Ech) is a dark-red pigment isolated from the sea urchin spines, shells, and ova. It has antioxidant, antimicrobial, and anti-inflammatory properties, but whether it can be used in asthma treatment has yet to be investigated. In this research, we aimed to study the inhibitory actions of Ech on allergic asthma symptoms in mice. Mice were divided into 4 groups (n = 8 for each): control, ovalbumin-challenged, and Ech-treated (0.1 and 1 mg/kg). At the end of the experiment, nasal scratching, lung oxidative stress, airway inflammation, and remodeling were assessed. In ovalbumin-challenged BALB/C mice, treatment with Ech significantly decreased nasal scratching, lung oxidative stress, inflammatory cell infiltration, mucus hyperproduction and hyperplasia of goblet cells, IgE levels, and inflammatory cytokines. It also inhibited NF-κB phosphorylation. This is the first study to investigate the immunomodulatory effect of Ech against allergic asthma in mice. According to our findings, we imply that Ech may be utilized as a treatment for allergic asthma.

Echinochrome Ameliorates Physiological, Immunological, and Histopathological Alterations Induced by Ovalbumin in Asthmatic Mice by Modulating the Keap1/Nrf2 Signaling Pathway.

Asthma is a persistent inflammatory disease of the bronchi characterized by oxidative stress, airway remodeling, and inflammation. Echinochrome (Ech) is a dark-red pigment with antioxidant and anti-inflammatory activities. In this research, we aimed to investigate the effects of Ech against asthma-induced inflammation, oxidative stress, and histopathological alterations in the spleen, liver, and kidney in mice. Mice were divided into four groups (n = 8 for each): control, asthmatic, and asthmatic mice treated intraperitoneally with 0.1 and 1 mg/kg of Ech. In vitro, findings confirmed the antioxidant and anti-inflammatory activities of Ech. Ech showed antiasthmatic effects by lowering the serum levels of immunoglobulin E (IgE), interleukin 4 (IL-4), and interleukin 1ß (IL-1ß). It attenuated oxidative stress by lowering malondialdehyde (MDA) and nitric oxide (NO) contents and increasing reduced glutathione (GSH), superoxide dismutase (SOD), glutathione-s-transferase (GST), and catalase (CAT) in the liver, spleen, and kidney. Moreover, it protected asthma-induced kidney and liver functions by increasing total protein and albumin and decreasing aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatinine, urea, and uric acid levels. Additionally, it ameliorated histopathological abnormalities in the lung, liver, spleen, and kidney. Additionally, molecular docking studies were used to examine the interactions between Ech and Kelch-like ECH-associated protein 1 (Keap1). PCR and Western blot analyses confirmed the association of Ech with Keap1 and, consequently, the regulatory role of Ech in the Keap1-(nuclear factor erythroid 2-related factor 2) Nrf2 signaling pathway in the liver, spleen, and kidney. According to our findings, Ech prevented asthma and its complications in the spleen, liver, and kidney. Inhibition of inflammation and oxidative stress are two of echinochrome's therapeutic actions in managing asthma by modulating the Keap1/Nrf2 signaling pathway.

Protective immune potential of multiple antigenic peptide (MAP) constructs comprising peptides that are shared by several cysteine peptidases against Schistosoma mansoni infection in mice.

In vaccine trials, Schistosoma mansoni cathepsin B1 (SmCB1), helminth cathepsins of the L family (e.g., SmCL3), and papain consistently induce highly significant reductions in challenge worm burden and egg viability, but generated no additive protective effects when used in combination. The protective capacity of the cysteine peptidases is associated with modest (SmCB1) and poor (cathepsins L) production of cytokines and antibodies, essentially of the type 2 axis, and is only marginally reduced upon use of proteolytically inactive enzymes. In this work, peptides shared by SmCB1, cathepsins of the L family, papain and other allergens were selected, synthesized as tetrabranched multiple antigen peptide constructs (MAP-1 and MAP-2), and used in two independent experiments to immunize outbred mice, in parallel with papain. The two peptides elicited significant (P < 0.05) reduction in challenge worm burden when compared to unimmunized mice, albeit lower than that achieved by papain. Protection was associated with modest serum type 2 cytokines and antibody levels in MAP-, and papain-immunized mice. Immunization with papain also elicited a reduction in parasite egg load, viability, and granuloma numbers in liver and intestine. MAP-1 and MAP-2 immunogens displayed some opposite effects- MAP-1 leading to higher egg numbers with poor vitality, whereas MAP-2 immunization yielded fewer eggs. Cysteine peptidase thus appear to carry peptides that elicit opposing outcomes, highlighting the difficulty of reaching fully fledged protection, unless a vaccine is based on carefully selected peptides and combined with an effective adjuvant.

A comprehensive and critical overview of schistosomiasis vaccine candidates.

A digenetic platyhelminth Schistosoma is the causative agent of schistosomiasis, one of the neglected tropical diseases that affect humans and animals in numerous countries in the Middle East, sub-Saharan Africa, South America and China. Several control methods were used for prevention of infection or treatment of acute and chronic disease. Mass drug administration led to reduction in heavy-intensity infections and morbidity, but failed to decrease schistosomiasis prevalence and eliminate transmission, indicating the need to develop anti-schistosome vaccine to prevent infection and parasite transmission. This review summarizes the efficacy and protective capacity of available schistosomiasis vaccine candidates with some insights and future prospects.

Does Curcumin Have a Role in the Interaction between Gut Microbiota and Schistosoma mansoni in Mice?

There is strong correlation between changes in abundance of specific bacterial species and several diseases including schistosomiasis. Several studies have described therapeutic effects of curcumin (CUR) which may arise from its regulative effects on intestinal microbiota. Thus, we examined the impact of CUR on the diversity of intestinal microbiota with/without infection by Schistosoma mansoni cercariae for 56 days. Enterobacteriaceae was dominating in a naive and S. mansoni infected mice group without CUR treatment, the most predominant species was Escherichia coli with relative density (R.D%) = 80.66% and the least one was Pseudomonas sp. (0.52%). The influence of CUR on murine microbiota composition was examined one week after oral administration of high (40) and low (20 mg/kg b.w.) CUR doses were administered three times, with two day intervals. CUR induced high variation in the Enterobacteriaceae family, characterized by a significant (p < 0.001) reduction in E. coli and asignificant (p < 0.001) increase in Pseudomonas sp. in both naïve and S. mansoni-infected mice, compared to untreated mice, in a dose-dependent manner. Additionally, our study showed the effects of high CUR doses on S. mansoni infection immunological and parasitological parameters. These data support CUR's ability to promote Pseudomonas sp. known to produce schistosomicidal toxins and offset the sequelae of murine schistosomiasis.

In vitro effect of curcumin on Schistosoma species viability, tegument ultrastructure and egg hatchability.

Schistosomiasis remains a severe problem of public health in developing countries. The development of resistance to praziquantel (PZQ) has justified the search for new alternative chemotherapies with new formulations, more effective, and without adverse effects. Curcumin (CUR), the major phenolic compound present in rhizome of turmeric (Curcuma longa L.), has been traditionally used against various diseases including parasitic infections. Here, the antischistosomal activity of CUR (50-500 µM), evaluated in parallel against S. mansoni and S. haematobium adult worms, appeared significant (P < 0.05 to < 0.0001) in a time- and dose-dependent manner. Two h incubation with CUR (500 µM) caused 100% irreversible killing of both schistosomal species. CUR (250 µM) caused the death of S. haematobium and S. mansoni worms after 2 h and 4 h, respectively. As CUR concentration decreases (50 µM), all coupled adult worms were separated into individual male and female but the worms remained viable up to 4 h. Scanning and transmission electron microscopy revealed that S. haematobium are more sensitive than S. mansoni to CUR schistosomicidal effects. In support, CUR was found to affect the antigenicity of surface membrane molecules of S. haematobium, but not S. mansoni. Of importance, CUR significantly (P < 0.05 to < 0.0001) affected S. mansoni eggs hatchability and viability, a ground for its use in chemotherapy of schistosomiasis mansoni and japonicum because of its increased bioavailability in the gastrointestinal tract. The data together emphasize that CUR is a promising potential schistosomicidal drug.

Hormonal-receptor positive breast cancer: IL-6 augments invasion and lymph node metastasis via stimulating cathepsin B expression.

Hormonal-receptor positive (HRP) breast cancer patients with positive metastatic axillary lymph nodes are characterized by poor prognosis and increased mortality rate. The mechanisms by which cancer cells invade lymph nodes have not yet been fully explored. Several studies have shown that expression of IL-6 and the proteolytic enzyme cathepsin B (CTSB) was associated with breast cancer poor prognosis. In the present study, the effect of different concentrations of recombinant human IL-6 on the invasiveness capacity of HRP breast cancer cell line MCF-7 was tested using an in vitro invasion chamber assay. The impact of IL-6 on expression and activity of CTSB was also investigated. IL-6 treatment promoted the invasiveness potential of MCF-7 cells in a dose-dependent manner. Furthermore, MCF-7 cells displayed elevated CTSB expression and activity associated with loss of E-cadherin and upregulation of vimentin protein levels upon IL-6 stimulation. To validate these results in vivo, the level of expression of IL-6 and CTSB in the carcinoma tissues of HRP-breast cancer patients with positive and negative axillary metastatic lymph nodes (pLNs and nLNs) was assessed. Western blot and immunohistochemical staining data showed that expression of IL-6 and CTSB was higher in carcinoma tissues in HRP-breast cancer with pLNs than those with nLNs patients. ELISA results showed carcinoma tissues of HRP-breast cancer with pLNs exhibited significantly elevated IL-6 protein levels by approximately 2.8-fold compared with those with nLNs patients (P < 0.05). Interestingly, a significantly positive correlation between IL-6 and CTSB expression was detected in clinical samples of HRP-breast cancer patients with pLNs (r = 0.78, P < 0.01). Collectively, this study suggests that IL-6-induced CTSB may play a role in lymph node metastasis, and that may possess future therapeutic implications for HRP-breast cancer patients with pLNs. Further studies are necessary to fully identify IL-6/CTSB axis in different molecular subtypes of breast cancer.

SCHISTOSOMA-SPECIFIC 26 KDA PROTEIN FOR A DIAGNOSIS OF ACUTE AND CHRONIC SCHISTOSOMIASIS.

One of the world wide major public health problems is the schistosomiasis that is caused by Schistosoma (S.) heamatobium. It is also one of the main concerns for the public health community in Egypt. There are several immunodiagnostic methods used for that diagnosis of such disease, but some are more sensitive and specific than others. The purified 26 kDa Schistosoma-specific protein (PSPA-26) detection in serum samples is found out to be more valuable in diagnosis; it also helps in the early diagnosis which will lead to the early treatment before the irreversible damage takes place. PSPA-26 was purified from whole worms by DEAE-Sephadex G-75 ion exchange chromatography and then was injected into rabbits to produce specific polyclonal antibodies (anti-PSPA-26 pAb) which were then used as a primary capture in the indirect ELISA technique to reveal its reactivity using infected human sera. The anti-PSPA-26 was then labeled with horse-radish peroxidase (HRP) and used as a secondary capture. Sandwich ELISA was done for serum samples of human and hamsters infected with S. heamatobium. The results revealed a sensitivity of 85% for human and 80% for hamster's samples, and a specificity of 95% for human and 91.1% for hamsters samples by comparing them with those infected with other parasites and control samples. Data obtained concluded that PSPA-26 antigen can be used as a diagnostic marker for S. haematobium infection using the sandwich ELISA which is cost effective and applicable technique.

SANDWICH-ELISA DEVELOPMENT FOR THE DIAGNOSIS OF TOXOPLASMA GONDI.

Toxoplasmosis is one of the most world-wide spread zoonosis representing a very serious clinical and veterinary problem. Although rare, congenital toxoplasmosis can cause severe ne- urological or ocular disease (leading to blindness) as well as cardiac and cerebral anomalies. Prenatal care must include education about prevention of toxoplasmosis. Thus, a standardized and approachable diagnostic tool for the serodiagnosis of toxoplasmosis is still needed. Serological tests are the most widely used biological tools for the diagnosis of toxoplasmosis worldwide. Sandwich-ELISA is a solid phase diagnostic method for detection of antigen or antibody that is used widely for diagnosis of protozoan and metazoan diseases of human and animals. In the present study, T. gondii SAG2 antigen was early detected in patient sera using Ssndwich-ELISA, PAb was prepared from anti-rabbit sera and used for coating and as conjugate in Sandwich-ELISA technique. 46 patients out of 50 were positive to Toxoplsama spp. with sensitivity and specificity 92% and 90%, respectively. The PPV was 90.2% and NPV was 83.3%. Finally, the result of our study showed that the Sandwich-ELISA designed in our study is easy to perform, not expensive, safe, and simple with good sensitivity and specificity.

DOT-ELISA AS A FIELD TEST FOR HYDATID DIAGNOSIS.

Cystic hydatid disease (Hydatidosis) is one of the most important parasitic zoonoses and remains a public health and economic problem all over the world. Cyst fluid was obtained from hepatic and pulmonary cysts for demonstration of protoscolices and hooklets. Therefore, a standardized and approachable diagnostic tool for the serodiagnosis of CE is still needed. Dot-ELISA is a solid phase diagnostic method for detection of antigen or antibody that is used widely for diagnosis of protozoan and metazoan diseases of human and animals. In the present study, E. granulosus protoscolex antigen was early detected in patient sera using Dot-ELISA, PAb was prepared from anti-rabbit sera and used for coating and as conjugate in Dot-ELISA technique. 48 patients out of 50 were positive to E. spp. with sensitivity and specificity 96% and 94%, respectively. The PPV was 94% and NPV was 90%. Finally, the present results showed that the Dot-ELISA was easy to perform, not expensive, safe, and with good sensitivity and specificity.

Immunodiagnosis of Egyptian human fasciolosis gigantica using Fas1 and Fas2 cysteine proteinase antigens.

Fasciolosis caused by Fasciola gigantica is one of the major public health problems in the world including Egypt. Immunodiagnostic methods are more applicable for their better sensitivity and specificity than other methods. The present study was conducted to cysteine proteinase (CP) antigens of F. gigantica in IgG-ELISA to diagnose human fasciolosis. IgG-ELISA with 2 cysteine proteinases of 27 kDa (Fas1) and 29 kDa (Fas2), obtained from the regurgitated materials of adult worms, were evaluated using serum samples from 90 Egyptian patients infected with F. gigantica, 55 patients with other parasitic infections and 50 healthy volunteers. The diagnostic sensitivity and specificity of Fas1 for detection of F. gigantica infection were 91.1% and 89.1%, respectively. The positivity of the assay was 95%. The positive and negative predicted values were 91% and 86%, respectively. These data suggest that IgG-ELISA with Fas1 is highly sensitive and specific assay and could be used for the immunodiagnosis of human fasciolosis.

In vitro and in vivo activities of arachidonic acid against Schistosoma mansoni and Schistosoma haematobium.

The development of arachidonic acid (ARA) for treatment of schistosomiasis is an entirely novel approach based on a breakthrough discovery in schistosome biology revealing that activation of parasite tegument-bound neutral sphingomyelinase (nSMase) by unsaturated fatty acids, such as ARA, induces exposure of parasite surface membrane antigens to antibody binding and eventual attrition of developing schistosomula and adult worms. Here, we demonstrate that 5 mM ARA leads to irreversible killing of ex vivo 1-, 3-, 4-, 5-, and 6-week-old Schistosoma mansoni and 9-, 10-, and 12-week-old Schistosoma haematobium worms within 3 to 4 h, depending on the parasite age, even when the worms were maintained in up to 50% fetal calf serum. ARA-mediated worm attrition was prevented by nSMase inhibitors, such as CaCl(2) and GW4869. Scanning and transmission electron microscopy revealed that ARA-mediated worm killing was associated with spine destruction, membrane blebbing, and disorganization of the apical membrane structure. ARA-mediated S. mansoni and S. haematobium worm attrition was reproduced in vivo in a series of 6 independent experiments using BALB/c or C57BL/6 mice, indicating that ARA in a pure form (Sigma) or included in infant formula (Nestle) consistently led to 40 to 80% decrease in the total worm burden. Arachidonic acid is already marketed for human use in the United States and Canada for proper development of newborns and muscle growth of athletes; thus, ARA has potential as a safe and cost-effective addition to antischistosomal therapy.

Innate immunogenicity and in vitro protective potential of Schistosoma mansoni lung schistosomula excretory--secretory candidate vaccine antigens.

Excretory-secretory products (ESP) of Schistosoma mansoni developing larvae are ideal potential vaccines as such molecules may readily induce host primary immune responses, and local memory immune response effectors that would target, surround, and pursue the larvae while negotiating the lung blood capillaries. We herein characterized the cytokines response ESP, e.g., SG3PDH, 14-3-3-like protein, TPX, and calpain induce in the natural context of infection, and defined the global cytokine profile conducive to effective schistosome larvae killing. Accordingly, spleen cells (SC) taken from naïve, and 7-, or 9-day S. mansoni-infected mice were stimulated in vitro with the selected ESP, in a recombinant or multiple antigen peptide (MAP) form, and examined for production of T helper type (Th) 1, Th2, and Th17 cytokines, and the ability to mediate in vitro attrition of lung-stage schistosomula. The study indicated that larval ESP principally elicit Th1 and Th17 type cytokines. Recombinant SG3PDH was the only test ESP to additionally activate SC from S. mansoni-infected BALB/c mice to release higher IL-4 levels than unstimulated SC and mediate significant (P < 0.0001) in vitro attrition of lung-stage larvae. Thus, our data suggested that a balance between Th1, Th17, and Th2 cytokines is required for effective schistosome larval elimination.

Impact of interleukin-1 and interleukin-6 in murine primary schistosomiasis.

BACKGROUND: Immunization with schistosome antigens invariably elicits a plethora of cytokines and, hence, it is reasonable to assume that these cytokines influence host responses to challenge lung-stage larvae and, consequently, the adult worm burden, and may be responsible for the erratic data generally observed in protection studies against schistosome infection. METHODS: Schistosoma mansoni-infected mice were administered with recombinant interleukin (IL)-1beta or IL-6 to evaluate the impact of cytokines in host responses to lung-stage schistosomula, and subsequent effects on adult worm parameters. Plasma lipid levels were assayed by colorimetric enzymatic tests and antibody responses by ELISA. Cytokine profile in peripheral blood mononuclear cells was evaluated by RT-PCR. RESULTS: S. mansoni infection elicited, at the time of parasite residency in the lung, significant increase in free fatty acids (FA) and decrease in cholesterol plasma levels in C57BL/6 and CD1 mice, and stimulation of mRNA expression for cytokines of T helper type (Th) 2 in BALB/c, Th1 in C57BL/6, and Th1/Th2 in CD1 mice. However, no specific antibody production was evident in any mouse strain. In BALB/c mice, exogenous IL-1beta-related plasma free FA level significant increase, stimulation of expression of IL-1 and IL-12 mRNA, and considerable increase in percent of specific antibody-producing mice were associated with significant reduction in adult worm burden and egg load. In contrast, exogenous IL-1beta elicited decrease in free FA plasma levels, and down-regulation of cytokines' mRNA expression in C57BL/6 and CD1 mice, changes associated with aggravation of the worm burden. Likewise, exogenous IL-6 failed to stimulate increase in plasma free FA levels or percent of antibody-producing mice except in BALB/c mice, effects that were protective for the host in BALB/c and for the parasite in C57BL/6 and CD1 mice. CONCLUSION: These findings were discussed in relation to the erratic data of protection experiments with schistosome subunit antigens in different mouse strains.

Influence of interleukin-2 and interferon-gamma in murine schistosomiasis.

Schistosoma mansoni-infected mice were administered at the time of parasite residency in the lung with recombinant murine interleukin (IL)-2 or interferon-gamma (IFN-gamma), to evaluate the impact of cytokines in host responses to primary schistosomiasis. S. mansoni lung-stage schistosomula did not affect plasma lipids levels in BALB/c, while elicited significant (p<0.05) increase in free fatty acids (FA) and decrease in cholesterol plasma levels in C57BL/6 and CD1 mice, and stimulated expression of mRNA for Th2 cytokines in BALB/c and Th1 cytokines in C57BL/6 and CD1 mice. Production of specific antibodies was negligible in the 3 strains. Interleukin-2 treatment elicited significant (p<0.001) decrease in triglycerides (TG) in CD1, and decrease in TG and cholesterol plasma levels and down-regulation of TNF-alpha mRNA expression in C57BL/6 mice. Induction of type 2 cytokines and/or IFN-gamma mRNA expression did not lead to increase in percentage of specific antibody responders in any mouse strain. Exogenous IL-2-related reduction in cholesterol plasma levels and TNF-alpha mRNA expression in C57BL/6 mice was associated with significant (p<0.05) decrease in adult worm recovery and egg count. Treatment with IFN-gamma elicited significant (p<0.05) free FA plasma levels increase in BALB/c and C57BL/6 and decrease in CD1 mice. Expression of type 2 cytokines mRNA was stimulated in BALB/c and CD1 mice, yet was not accompanied with increase in humoral responses. Exogenous IFN-gamma-related reduction in free FA plasma levels and IFN-gamma mRNA response, and up-regulation of TNF-alpha mRNA expression in CD1 mice were associated with significant increase in adult worm burden and egg load. The data were discussed in an attempt to define host factors predictive of resistance to schistosome infection.