1.Protective and Anti-Pathology Effects of Sm Fructose-1,6-Bisphosphate Aldolase-Based DNA Vaccine against Schistosoma mansoni by Changing Route of Injection.
Mohamed SABER ; Tarek DIAB ; Olft HAMMAM ; Amr KARIM ; Amina MEDHAT ; Mamdouh KHELA ; Ehab EL-DABAA
The Korean Journal of Parasitology 2013;51(2):155-163
This study aimed to evaluate the efficacy of fructose-1,6-bis phosphate aldolase (SMALDO) DNA vaccination against Schistosoma mansoni infection using different routes of injection. The SMALDO has been cloned into the eukaryotic expression vector pcDNA3.1/V5-His TOPO-TA and was used in injecting Swiss albino mice intramuscularly (IM), subcutaneously (SC), or intraperitoneally (IP) (50 microg/mouse). Mice vaccinated with non-recombinant pcDNA3.1 served as controls. Each group was immunized 4 times at weeks 0, 2, 4, and 6. Two weeks after the last booster dose, all mice groups were infected with 80 S. mansoni cercariae via tail immersion. At week 8 post-infection, animals were sacrificed for assessment of parasitological and histopathological parameters. High anti-SMALDO IgG antibody titers were detected in sera of all vaccinated groups (P<0.01) compared to the control group. Both the IP and SC vaccination routes resulted in a significant reduction in worm burden (46.2% and 28.9%, respectively, P<0.01). This was accompanied by a significant reduction in hepatic and intestinal egg counts (41.7% and 40.2%, respectively, P<0.01) in the IP group only. The number of dead eggs was significantly increased in both IP and IM groups (P<0.01). IP vaccination recorded the highest significant reduction in granuloma number and diameter (54.7% and 29.2%, respectively, P<0.01) and significant increase in dead miracidia (P<0.01). In conclusion, changing the injection route of SMALDO DNA vaccination significantly influenced the efficacy of vaccination. SMALDO DNA vaccination via IP route could be a promising protective and anti-pathology vaccine candidate against S. mansoni infection.
Animals
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Antibodies, Helminth/blood
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Disease Models, Animal
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Female
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Fructose-Bisphosphate Aldolase/genetics/*immunology
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Histocytochemistry
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Immunoglobulin G/blood
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Injections, Intramuscular
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Injections, Intraperitoneal
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Injections, Subcutaneous
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Mice
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Parasite Load
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Schistosoma mansoni/enzymology/genetics/*immunology
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Schistosomiasis mansoni/immunology/parasitology/pathology/*prevention & control
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Vaccination/methods
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Vaccines, DNA/administration & dosage/genetics/*immunology
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Vaccines, Synthetic/administration & dosage/genetics/immunology
2.Protective Role of Purified Cysteine Proteinases against Fasciola gigantica Infection in Experimental Animals.
Eman EL-AHWANY ; Ibrahim RABIA ; Faten NAGY ; Mona ZOHEIRY ; Tarek DIAB ; Suher ZADA
The Korean Journal of Parasitology 2012;50(1):45-51
Fascioliasis is one of the public health problems in the world. Cysteine proteinases (CP) released by Fasciola gigantica play a key role in parasite feeding, migration through host tissues, and in immune evasion. There has been some evidence from several parasite systems that proteinases might have potential as protective antigens against parasitic infections. Cysteine proteinases were purified and tested in vaccine trials of sheep infected with the liver fluke. Multiple doses (2 mg of CP in Freund's adjuvant followed by 3 booster doses 1 mg each at 4 week intervals) were injected intramuscularly into sheep 1 week prior to infect orally with 300 F. gigantica metacercariae. All the sheep were humanely slaughtered 12 weeks after the first immunization. Changes in the worm burden, ova count, and humoral and cellular responses were evaluated. Significant reduction was observed in the worm burden (56.9%), bile egg count (70.7%), and fecel egg count (75.2%). Immunization with CP was also found to be associated with increases of total IgG, IgG1, and IgG2 (P<0.05). Data showed that the serum cytokine levels of pro-inflammatory cytokines, IL-12, IFN-gamma, and TNF-alpha, revealed significant decreases (P<0.05). However, the anti-inflammatory cytokine levels, IL-10, TGF-beta, and IL-6, showed significant increases (P<0.05). In conclusion, it has been found that CP released by F. gigantica are highly important candidates for a vaccine antigen because of their role in the fluke biology and host-parasite relationships.
Animals
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Antibodies, Helminth/immunology
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Cysteine Proteases/administration & dosage/*immunology/isolation & purification
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Cytokines/immunology
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Fasciola/chemistry/*enzymology/immunology
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Fasciola hepatica/immunology/physiology
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Fascioliasis/immunology/parasitology/*prevention & control
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Female
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Helminth Proteins/administration & dosage/*immunology/isolation & purification
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Humans
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Male
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Protective Agents/*administration & dosage/isolation & purification
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Sheep
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Vaccines/immunology
3.Monoclonal Antibody-Based Dipstick Assay: A Reliable Field Applicable Technique for Diagnosis of Schistosoma mansoni Infection Using Human Serum and Urine Samples.
Zeinab DEMERDASH ; Salwa MOHAMED ; Mohamed HENDAWY ; Ibrahim RABIA ; Mohy ATTIA ; Zeinab SHAKER ; Tarek M DIAB
The Korean Journal of Parasitology 2013;51(1):93-98
A field applicable diagnostic technique, the dipstick assay, was evaluated for its sensitivity and specificity in diagnosing human Schistosoma mansoni infection. A monoclonal antibody (mAb) against S. mansoni adult worm tegumental antigen (AWTA) was employed in dipstick and sandwich ELISA for detection of circulating schistosome antigen (CSA) in both serum and urine samples. Based on clinical and parasitological examinations, 60 S. mansoni-infected patients, 30 patients infected with parasites other than schistosomiasis, and 30 uninfected healthy individuals were selected. The sensitivity and specificity of dipstick assay in urine samples were 86.7% and 90.0%, respectively, compared to 90.0% sensitivity and 91.7% specificity of sandwich ELISA. In serum samples, the sensitivity and specificity were 88.3% and 91.7% for dipstick assay vs. 91.7% and 95.0% for sandwich ELISA, respectively. The diagnostic efficacy of dipstick assay in urine and serum samples was 88.3% and 90.0%, while it was 90.8% and 93.3% for sandwich ELISA, respectively. The diagnostic indices of dipstick assay and ELISA either in serum or in urine were statistically comparable (P>0.05). In conclusion, the dipstick assay offers an alternative simple, rapid, non-invasive technique in detecting CSA or complement to stool examinations especially in field studies.
Animals
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Antibodies, Helminth/diagnostic use/isolation & purification
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Antibodies, Monoclonal/diagnostic use/isolation & purification
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Antigens, Helminth/*blood/*urine
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Diagnostic Tests, Routine/*methods
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Humans
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Immunoassay/methods
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Parasitology/*methods
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*Point-of-Care Systems
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Schistosoma mansoni/immunology/*isolation & purification
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Schistosomiasis mansoni/*diagnosis
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Sensitivity and Specificity