Expression profile, localization of an 8-kDa calcium-binding protein from Schistosoma japonicum (SjCa8), and vaccine potential of recombinant SjCa8 (rSjCa8) against infections in mice

Lv, Ziquan; Yang, Linlin; Hu, Shaomin; Sun, Xi; He, Hailong; He, Shao-yun; Li, Zhengyu; Zhou, Yanping; Fung, Ming Chiu; Yu, Xinbing; Zheng, Haixue; Ai-lian, Cao; Wu, Zhongdao

doi:10.1007/s00436-008-1249-0

Cited by 21 publications

(19 citation statements)

References 34 publications

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“…Purification was done by Ni-NTA under denaturing condition. His-tag was employed in the purification of recombinant proteins based on immobilized metal affinity chromatography, which has been used in a number of studies (Kim et al 2007;Lv et al 2009). The result indicated that the above mentioned conditions were appropriate for the purification and refolding of apyrase protein.…”

Section: Discussionmentioning

confidence: 99%

Cloning, expression, and characterization of salivary apyrase from Aedes albopictus

Dong

et al. 2011

Parasitol Res

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Apyrases (ATP diphosphohydrolase) hydrolyze the phosphodiester bonds of nucleoside tri- and diphosphates to orthophosphate and mononucleodides. They can inhibit platelet activation by depletion of adenosine diphosphate. In the current study, the Escherichia coli expression vector pET-19b equipped with an N-terminal histidine tag was applied to express the apyrase of Aedes albopictus. The gene-coding mature apyrase protein was amplified by RT-PCR and cloned into pET-19b. Soluble apyrase protein with high purity was successfully obtained by utilization of the suitable renaturation approach and Ni-NTA purification column. Four monoclonal antibodies to apyrase from A. albopictus were produced in male BALB/c mice immunized with the renatured apyrase. Using immunofluorescence assay and immunoblotting analysis, recombinant apyrase showed fine consistency with native apyrase. From kinetic analysis, it had a K (m) of 11.6 μM and V (max) of 1.02 nM/S/μg protein for adenosine triphosphate. Adenosine diphosphate-induced platelet aggregation was inhibited by approximately 6% when 0.4 μM recombinant apyrase was added and by about 9.5% when the concentration of recombinant apyrase was 0.8 μM. The effect on platelet aggregation was dose dependent. In conclusion, the apyrase of A. albopictus was cloned and expressed highly in the E. coli expression system. Recombinant apyrase protein showed biological activity, and anti-apyrase monoclonal antibody was also prepared.

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Section: Discussionmentioning

confidence: 99%

Cloning, expression, and characterization of salivary apyrase from Aedes albopictus

Dong

et al. 2011

Parasitol Res

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“…The effect of linalool on integrity of the cercarial tegument was evaluated using detection of tegumentary protein SjCa8, a cercariae specific molecule, which has been described by Lv et al [19]. Briefly, after treatment with different compound for 30 min, and subsequent three washes (5 min each) with 1 × PBS, cercariae were incubated for 45 min at 37°C in PBS containing 5% bovine serum albumin (BSA, Sigma).…”

Section: Methodsmentioning

confidence: 99%

“…Briefly, after treatment with different compound for 30 min, and subsequent three washes (5 min each) with 1 × PBS, cercariae were incubated for 45 min at 37°C in PBS containing 5% bovine serum albumin (BSA, Sigma). The worms were treated for 45 min at 37°C with the primary antibodies (1:500 dilution) to recombinant SjCa8 prepared in our laboratory [19]. After an additional washing step, parasites were incubated for 1 h at room temperature with Cy3-conjugated anti-mouse IgG diluted at 1:100, washed thrice and then observed under an FV 500-IX81 laser scanning confocal system (Olympus, Japan) with Krypton/Argon laser and appropriate filters.…”

Section: Methodsmentioning

confidence: 99%

Linalool, derived from Cinnamomum camphora (L.) Presl leaf extracts, possesses molluscicidal activity against Oncomelania hupensis and inhibits infection of Schistosoma japonicum

et al. 2014

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BackgroundSchistosomiasis japonicum remains a considerable economic and public health concern in China, the Philippines and Indonesia. Currently available measures to control the unique intermediate host Oncomelania hupensis are frequently associated with severe side effects. Previous studies have demonstrated that linalool-rich extracts from various plants exhibited promising biological activities including cytotoxic, anti-microbial and anti-parasitic properties.MethodsWe identified the components of leaf extracts from Cinnamomum camphora by gas chromatography coupled to mass spectrometry (GC-MS) and investigated molluscicidal and larvicidal effects of linalool against O. hupensis and Schistosoma japonicium. The ultrastructural alterations in gills, salivary gland, stomach and hepatopancreas of snails were observed under the light microscope and transmission electron microscope, and lesions to tegument of cercaria were examined under a light microscope and fluorescence microscope. We then evaluated the effects of linalool on skin penetration and migration of schistosomula and adult survival by measurement of worm burden and egg counts in Balb/C mice infected with linalool-treated cercariae.ResultsIn the present work, 44 components were identified from the leaf extracts of C. camphora, of which linalool was the most abundant constituent. Linalool exhibited the striking molluscicidal and larvicidal effects with LC50 = 0.25 mg/L for O. hupensis and LC50 = 0.07 mg/L for cercaria of S. japonicium. After exposure to linalool, damage to the gills and hepatopancreas of the snails, and to the tegument and body-tail joint of cercariae was apparent. In addition, linalool markedly reduced the recovered schistosomulum from mouse skin after challenge infection, and therefore decreased the worm burden in infected animals, but not fecundity of female adults of the parasite.ConclusionsOur findings indicated that linalool might be a novel chemotherapeutic agent against S. japonicium and the snail intermediate host.

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“…One week after the final booster dose, mice were bled, and the antiserum was collected. Blood samples from C57BL/6 mouse infected with S. japonicum and normal mouse serum were prepared as our previous study (Lv et al 2009). All sera were separated by centrifuging the blood samples at 4°C (10,000 rpm, 20 min) and stored at −70°C until use.…”

Section: Mouse Sera Preparationmentioning

confidence: 99%

Cloning, molecular characterization of a 13-kDa antigen from Schistosoma japonicum, Sj13, a putative salivary diagnosis candidate for Schistosomiasis japonica

Zhou

Sun

et al. 2009

Parasitol Res

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Saliva has been suggested as an easily accessible and a noninvasive diagnostic alternative for detection of antibodies. To identify and characterize Schistosoma japonicum (S. japonicum) antigens that are recognized by saliva of infected host, we have used a pool of saliva from infected patients to immunoscreen an egg cDNA library of S. japonicum. The open reading frame of the isolated two clones encodes same protein of 116 amino acids exhibiting 100% identity to an amino acid sequence (AY222893) of S. japonicum in NCBInr database. The protein encoded is inferred a secretory protein with a molecular mass of 13 kDa (Sj13) and shares no homology to any entries in the NCBInr database, demonstrating that Sj13 might be a schistosome-specific protein. Reverse transcriptase polymerase chain reaction, Western blotting, and immunolocalization analysis revealed Sj13 could be detected in cercaria, adult, and egg and was localized to forehead and tegument of cercaria, cell body ("cytons") of adult worm, egg shell, and epidermal plate of miracidium. Furthermore, Sj13 showed a good antigenicity when reacted with saliva or serum from schistosomiasis patients. The recombinant Sj13 (rSj13) expressed and purified from Escherichia coli was applied to detect its specific salivary antibody for schistosomiasis diagnosis by an indirect enzyme linked immunosorbent assay (ELISA). Preliminary laboratory test of 116 subjects, 40 with parasitologically proven S. japonicumm infection, 46 with other infectious diseases, and 30 negative controls exhibited 92.50% sensitivity with saliva/rSj13 and 95.00% with serum/SWAP (P > 0.05). The specificity of the ELISA using saliva/rSj13 was 92.11% versus 85.53% with serum/SWAP (P < 0.05). No direct correlations of anti-Sj13 IgG levels with egg counts in stool were observed in saliva detection. These results suggest that Sj13 specific salivary antibody detection may be useful as an antigen for the salivary diagnosis of schistosomiasis japonica and contribute to epidemiological study of schistosomiasis infection in endemic areas.

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Expression profile, localization of an 8-kDa calcium-binding protein from Schistosoma japonicum (SjCa8), and vaccine potential of recombinant SjCa8 (rSjCa8) against infections in mice

Cited by 21 publications

References 34 publications

Cloning, expression, and characterization of salivary apyrase from Aedes albopictus

Cloning, expression, and characterization of salivary apyrase from Aedes albopictus

Linalool, derived from Cinnamomum camphora (L.) Presl leaf extracts, possesses molluscicidal activity against Oncomelania hupensis and inhibits infection of Schistosoma japonicum

Cloning, molecular characterization of a 13-kDa antigen from Schistosoma japonicum, Sj13, a putative salivary diagnosis candidate for Schistosomiasis japonica

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