Prophylactic antineoplastic activity of Toxoplasma gondii RH derived antigen against ehrlich solid carcinoma with evidence of shared antigens by comparative immunoblotting.

BACKGROUND: With cancer cases escalation, an urgent request to develop novel combating strategies arise. Pathogen-based cancer-immunotherapy is getting more consideration. Autoclaved parasitic antigens seem promising candidates, taking steadily their first steps. Our aim was to examine the prophylactic antineoplastic activity of autoclaved Toxoplasma vaccine (ATV) and to test for the shared antigen theory between Toxoplasma gondii and cancer cells. METHODS: Mice were immunized with ATV followed by Ehrlich solid carcinoma (ESC) inoculation. Tumor weight, volume, histopathology, and immunohistochemistry for CD8+ T cells, Treg cells and VEGF were assessed. In addition, the proposed shared antigen theory between parasites and cancer was also verified using SDS-PAGE and immunoblotting. RESULTS: Results revealed powerful prophylactic activity of ATV with 13.3% inhibition of ESC incidence, significant reduction in tumor weight and volume in ATV vaccinated mice. Immunologically, significantly higher CD8+T cells and lower FOXP3+ Treg cells surrounded and infiltrated ESC in ATV immunized mice with higher CD8+T/Treg cells ratio and significant antiangiogenic effect. Moreover, SDS-PAGE and immunoblotting showed four shared bands between Ehrlich carcinoma and ATV of approximate molecular weights 60, 26, 22 and 12.5 KDa. CONCLUSION: Exclusively, we demonstrated a prophylactic antineoplastic activity of autoclaved Toxoplasma vaccine against ESC. Moreover, to the best of our knowledge this is the first report highlighting the existence of cross-reactive antigens between Toxoplasma gondi parasite and cancer cells of Ehrlich carcinoma.

Toxoplasma gondii-derived antigen modifies tumor microenvironment of Ehrlich solid carcinoma murine model and enhances immunotherapeutic activity of cyclophosphamide.

Pathogen-based cancer vaccine is a promising immunotherapeutic weapon to stimulate cancer immunosuppressive state. Toxoplasma gondii is a potent immunostimulant, and low-dose infection was linked to cancer resistance. Our goal was to evaluate the therapeutic antineoplastic activity of autoclaved Toxoplasma vaccine (ATV) against Ehrlich solid carcinoma (ESC) in mice in reference to and in combination with low-dose cyclophosphamide (CP), a cancer immunomodulator. Mice inoculation with ESC was followed by applying different treatment modalities including ATV, CP, and CP/ATV. We evaluated the impact of the different treatments on liver enzymes and pathology, tumor weight, volume, and histopathological changes. Using immunohistochemistry, we evaluated CD8+ T cell, FOXP3+ Treg, CD8+/Treg outside and inside ESC, and angiogenesis. Results showed significant tumor weights and volumes reduction with all treatments with 13.3% inhibition of tumor development upon combined CP/ATV use. Significant necrosis and fibrosis were noted in ESC by all treatments with improved hepatic functions versus non-treated control. Although ATV was almost equivalent to CP in tumor gross and histopathology, it promoted an immunostimulatory activity with significant Treg cells depletion outside ESC and CD8+ T cells infiltration inside ESC with higher CD8+ T/Treg ratio inside ESC superior to CP. Combined with CP, ATV exhibited significant synergistic immunotherapeutic and antiangiogenic action compared to either treatment alone with significant Kupffer cells hyperplasia and hypertrophy. Exclusively, therapeutic antineoplastic and antiangiogenic activity of ATV against ESC was verified that boosted CP immunomodulatory action which highlights a novel biological cancer immunotherapeutic vaccine candidate.

Effect of alginate nanoparticles on the immunogenicity of excretory-secretory antigens against acute toxoplasmosis in murine model.

Efficacy of alginate nanoparticles (Alg-NPs) as vaccine delivery for the excretory-secretory antigens (ESAs) against the virulent strain of Toxoplasma gondii was evaluated. Swiss albino mice were intraperitoneally immunized with three doses of either in vivo and in vitro-prepared ESA vaccines, 20 µg each, at 2-week intervals, then were challenged with 2500 tachyzoites of the RH HXGPRT (-) Toxoplasma gondii strain, four weeks later. Mice mortality, tachyzoite number in both peritoneal fluid and impression smear, and viability, ultrastructural tachyzoite changes, measuring immunological markers, and histopathological changes of both liver and spleen were studied, in comparison to alum adjuvanted ESAs and infected control subgroups. The in vivo-prepared Alg-NPs loaded ESAs vaccinated subgroups induced significant reduction in mice mortality, tachyzoite count in both peritoneal fluid and impression smears and viability. Scanning electron microscopy revealed tachyzoites deformities with multiple irregularities, while transmission electron microscopy showed tachyzoites distortion, disrupted plasma membranes, loss of nuclear integrities, and absence of dense granules with extensive vacuolations. A statistically significant increase in the level of both IFN-γ and anti-Toxoplasma IgG was noted. Histopathological results recorded amelioration of the pathological changes induced by Toxoplasma infection in both liver and spleen, with scanty parasites. Therefore, Alg-NPs proved its effectiveness in enhancing the ESAs antigencity, and recommended to test its potentiality as drugs carrier for anti-Toxoplasma agents to enhance their therapeutic efficacy.

Protective effect of surface-modified berberine nanoparticles against LPS-induced neurodegenerative changes: a preclinical study.

Berberine (BBR) exerts documented protection against neurodegenerative disorders. However, data on the effect of nano-encapsulation on the neuroprotective effect of BBR are lacking. We investigated the effect of BBR loading into chitosan (CS) nanoparticles (NPs) and their surface modification with Tween 80 (T80), polyethylene glycol 4000 (PEG), and miltefosine (MFS) against lipopolysaccharide (LPS)-induced neurodegenerative changes in addition to hepatotoxicity in rats. BBR-NPs were prepared by ionic gelation and characterized for morphology by transmission electron microscopy (TEM), colloidal properties, and entrapment efficiency (EE%). The neuroprotective and hepatoprotective effects of a 14-day pretreatment with four BBR-NPs formulations (4 mg/kg BBR/day) by intraperitoneal (i.p.) injection were challenged by a single i.p. 4 mg/kg dose of LPS on the fifteenth day. Neuroprotective efficacy and potential toxicity of BBR-NPs relative to BBR solution were assessed biochemically and histopathologically. One-way ANOVA followed by Tukey's comparison test was used for statistical analysis. CS nano-encapsulation and surface modification of BBR-NPs altered the neuroprotective and hepatoprotective effects of BBR depending on the physicochemical and/or biological effects of BBR, CS, coating materials, and NP-related features. Similar to the prophylactic and treatment efficacy of NPs for brain delivery, safety of these nanostructures and their individual formulation components warrants due research attention.

Repositioning of chlorambucil as a potential anti-schistosomal agent.

As parasites and cancer cells share many lifestyle and behavioral resemblances, repositioning of anti-cancerous agents as anti-parasitic is quite trendy, especially those sharing the same therapeutic targets. Therefore, the current study investigated the in vitro efficacy of ascending concentrations of chlorambucil (0.5-20µg/ml) against adult Schistosoma mansoni worms, over 72h. Additionally, its in vivo effects against the different developmental stages of the worm were assessed, after an oral dose of 2.5mg/kg/day for five successive days, through evaluating the worm load reduction and worms' morphological alterations and oogram changes. In addition to tissue egg count, a histopathological study of the liver was conducted. In vitro, chlorambucil demonstrated noticeable anti-schistosomal effects in the form of progressive reductions of the worms' viability in a dose dependent manner. Complete worm death was achieved at 72h incubation with 5µg/ml drug concentration. In vivo, chlorambucil induced a significant reduction in the total worm load against all developmental stages. Its highest impact was evident against the juvenile stage, where it induced 75.8% total worm load reduction, and 89.2% and 86.7% intestinal and hepatic egg counts reduction, respectively, along with ogram alterations. Besides, it induced significant shortening of both male and female worms and promoted an amelioration of hepatic histopathology. Results show that chlorambucil possesses favorable in vitro and in vivo anti-schistosomal activity. The highest in vivo efficacy was against the juvenile stage of S. mansoni, significantly superior to praziquantel, with extended potency to the adult stage. Further studies are recommended for chlorambucil target verification and to enhance its therapeutic efficacy.

Anti-Arthritic Activity of Schistosoma mansoni and Trichinella spiralis Derived-Antigens in Adjuvant Arthritis in Rats: Role of FOXP3+ Treg Cells.

A growing body of evidence supports the concept of helminths therapy in a variety of autoimmune diseases. Here, we aimed to investigate the protective effects of autoclaved Schistosoma mansoni antigen (ASMA) and Trichinella spiralis antigen (ATSA) on the clinical and immunopathological features of rheumatoid arthritis (RA). Adjuvant arthritis was induced by subcutaneous and intradermal injections of complete Freund's adjuvant into the plantar surface of the right hind paw and the root of the tail, respectively. Rats were randomly assigned to serve as normal control, untreated arthritis, ASMA or ATSA-treated arthritis groups. Antigens were given by intradermal injection in two doses, two weeks apart. The development, progression of arthritic features, and the impact on animals' gait and body weight were followed up for 4 weeks. The associated changes in serum cytokines (IL-17, IFN-γ and IL-10), joints' histopathology and immunohistochemistry of Foxp3+ T regulatory cells (Tregs) were evaluated at the end of the study. Treatment with either ASMA or ATSA attenuated the progression of clinical features of polyarthritis, improved gait and body weight gain, reduced the elevated serum IL-17 and further increased both IFN-γ and IL-10. Histopathologically, this was associated with a remarkable regression of paws' inflammation that was limited only to the subcutaneous tissue, and a significant increase in the number of Foxp 3+ cells versus the untreated arthritis group. In conclusion, both Schistosoma mansoni and Trichinella spiralis derived antigens exerted protective effect against adjuvant arthritis with better effect achieved by ASMA treatment. This anti-arthritic activity is attributed to upregulation of the Foxp3+ Tregs, with subsequent favorable modulation of both pro- and anti-inflammatory cytokines. The use of autoclaved parasitic antigens excludes the deleterious effects of imposing helminthic infection by using live parasites, which may pave the way to a new therapeutic modality in treating RA.

Could miltefosine be used as a therapy for toxoplasmosis?

Toxoplasmosis is a zoonotic protozoal disease affecting more than a billion people worldwide. The shortfalls of the current treatment options necessitate the development of non-toxic and well-tolerated, efficient alternatives especially against the cyst form. The current study was undertaken to investigate, for the first time, the potential potency of miltefosine against Toxoplasma gondii infection in acute and chronic experimental toxoplasmosis. Results showed that there is no evidence of anti-parasitic activity of miltefosine against T. gondii tachyzoites in acute experimental toxoplasmosis. However, anti-parasitic activity of miltefosine against T. gondii cyst stage in chronic experimental toxoplasmosis could not be excluded as demonstrated by significant reduction in brain cyst burden. Moreover, considerable morphological changes in the cysts were revealed by light and electron microscopy study and also by amelioration of pathological changes in the brain. Future studies should focus on enhancement of anti-toxoplasma activity of miltefosine against chronic toxoplasmosis using formulation based nanotechnology. To the best of our knowledge, this is the first study highlighting efficacy of miltefosine against chronic toxoplasmosis, thus, increasing the list of diseases that can be targeted by this drug.

Immunohistochemical detection of HCV in nerves and muscles of patients with HCV associated peripheral neuropathy and myositis.

BACKGROUND: Chronic hepatitis C Virus (HCV) infection may be associated with numerous extrahepatic manifestations, such as mixed cryoglobulinaemia, membranoproliferative glomerulonephritis, sicca syndrome. Cryoglobulinaemia (CG) is a condition characterized by the presence of serum proteins that reversibly precipitate in the cold. The objective of the present work was to study the histopathological changes in neuromuscular biopsies in patients with HCV associated peripheral neuropathy, or myopathy; with and without cryoglobulinemia, and to assess the presence of HCV in nerve and muscle tissues of those patients which might clarify some pathogenetic mechanisms for neuropathy, and myopathy associated with HCV. METHODS: The study was conducted on 17 cases of HCV infected patients with peripheral neuropathy and myositis. All patients were subjected to thorough laboratory investigations, neurological examination, electrophysiologic studies including nerve conduction, and needle EMG studies. RESULTS: Histopathological examination of nerve biopsies showed features of vascultis in 2/10 cases, interstitial inflammatory infiltrates in 5/10. Muscle biopsies showed intense inflammatory reaction, degenerative changes in the muscles of 3/10 cases diagnosed as myositis. Immunohistochemical results, showed in nerve biopsies, 7/10 cases with positive reaction for HCV with nuclear and perinuclear staining.. Two patients showed positive reaction in the epineural, and endoneural blood vessels and a negative reaction in nerve bundles, while in five patients, reaction was only positive in the nerve bundles. In muscle biopsies, 7/10 cases showed positive reaction for HCV in the nuclei of the muscle fibers, including the cases presented with myositis. CONCLUSION: The presence of HCV particles in nerve and muscle biopsies of patients with peripheral neuropathy suggests a virus triggered immune mediated mechanism.