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Functionally Expression of Metalloproteinase in Taenia solium Metacestode and Its Evaluation for Serodiagnosis of Cysticercosis.

Zhang Y, Bae YA, Zong HY, Kong Y, Cai GB - Iran J Parasitol (2016 Jan-Mar)

Bottom Line: After refolding and purification, enzymatic properties of the recombinant metalloproteinase were observed.The recombinant TsMP protein showed proteolytic activity, which preferred host extracellular matrix proteins such as collagen and fibronectin as degradable substrates.TsMP might be involved in the processing of numerous host proteins and play an important role in the parasite life cycle.

View Article: PubMed Central - PubMed

Affiliation: Dept. of Medical Genetics, Wuhan University School of Basic Medicial Sciences, Wuhan 430071, China.

ABSTRACT

Background: Parasite proteases have important roles in cleavage of host proteins during the invasion of host tissues and participate in the parasite's evasion from the host's immune response. The aim of the present study was to estimate a metalloproteinase properties of Taenia solium metacestode (TsMP) during host-parasite interactions, and evaluate its potential as a serodiagnostic antigen for cysticercosis.

Methods: The cDNA coding for the mature catalytic domain of TsMP was cloned into pGEX-6P-1 expression vector. A recombinant glutathione S-transferase and TsMP fusion protein was induced. After refolding and purification, enzymatic properties of the recombinant metalloproteinase were observed. Immunoblot assay was processed to evaluate its potential as a serodiagnostic antigen for cysticercosis.

Results: The recombinant TsMP protein showed proteolytic activity, which preferred host extracellular matrix proteins such as collagen and fibronectin as degradable substrates. In immunoblot assay, 87.5% of sera from patients with cysticercosis showed strong reactivity. In sera from patients with other parasitic infections and from normal controls, it showed high specificity.

Conclusions: TsMP might be involved in the processing of numerous host proteins and play an important role in the parasite life cycle. A single recombinant TsMP antigen could have a potential value for serodiagnosis of cysticercosis.

No MeSH data available.


Related in: MedlinePlus

Immunoblot analysis of Taenia solium metacestode recombinant mTsMP protein against serum samples from patients with different parasitic infection. After transfer-blot of recombinant mTsMP separated by 12% SDS-PAGE, each strip was incubated with the respective serum in a dilution of 1:200, subsequently with 1:1,000 diluted peroxidase conjugated anti-human IgG. The colour reactions were developed with 4-chloro-1-naphthol (4C1N) and 0.03% hydrogen peroxide (H2O2). Panel TsM, cysticercosis; panel Ec, echinococcosis; panel Sp, sparganosis; panel Pw, paragonimiasis; panel Cs, clonorchiasis; panel Normal, healthy controls. Lanes 1-10, each represent serum from different patient. The low molecular weight marker in kilodaltons (kDa) was shown at the left
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Figure 5: Immunoblot analysis of Taenia solium metacestode recombinant mTsMP protein against serum samples from patients with different parasitic infection. After transfer-blot of recombinant mTsMP separated by 12% SDS-PAGE, each strip was incubated with the respective serum in a dilution of 1:200, subsequently with 1:1,000 diluted peroxidase conjugated anti-human IgG. The colour reactions were developed with 4-chloro-1-naphthol (4C1N) and 0.03% hydrogen peroxide (H2O2). Panel TsM, cysticercosis; panel Ec, echinococcosis; panel Sp, sparganosis; panel Pw, paragonimiasis; panel Cs, clonorchiasis; panel Normal, healthy controls. Lanes 1-10, each represent serum from different patient. The low molecular weight marker in kilodaltons (kDa) was shown at the left

Mentions: To assess the diagnostic value of recombinant mTsMP, we tested its immunoreactivity by an immunoblot assay using individual sera from patients with various helminthic infections including cysticercosis, echinococcosis, sparganosis, paragonimiasis and clonorchiasis. A typical result of the immunoblot analysis is shown in Fig. 5. A strong recognition of recombinant mTsMP was observed in 87.5% (35/40 cases) of sera from patients with cysticercosis who were examined (Fig. 5, panel TsM). Meanwhile, weak cross-reactivity was observed in only 1 sample from a patient with echinococcosis (Fig. 5, panel Ec). None of the sera from patients with other parasitic infections or from normal controls showed any antibody positive reactivity (Fig. 5, panel Sp, Pw, Cs and Normal).


Functionally Expression of Metalloproteinase in Taenia solium Metacestode and Its Evaluation for Serodiagnosis of Cysticercosis.

Zhang Y, Bae YA, Zong HY, Kong Y, Cai GB - Iran J Parasitol (2016 Jan-Mar)

Immunoblot analysis of Taenia solium metacestode recombinant mTsMP protein against serum samples from patients with different parasitic infection. After transfer-blot of recombinant mTsMP separated by 12% SDS-PAGE, each strip was incubated with the respective serum in a dilution of 1:200, subsequently with 1:1,000 diluted peroxidase conjugated anti-human IgG. The colour reactions were developed with 4-chloro-1-naphthol (4C1N) and 0.03% hydrogen peroxide (H2O2). Panel TsM, cysticercosis; panel Ec, echinococcosis; panel Sp, sparganosis; panel Pw, paragonimiasis; panel Cs, clonorchiasis; panel Normal, healthy controls. Lanes 1-10, each represent serum from different patient. The low molecular weight marker in kilodaltons (kDa) was shown at the left
© Copyright Policy
Related In: Results  -  Collection

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Figure 5: Immunoblot analysis of Taenia solium metacestode recombinant mTsMP protein against serum samples from patients with different parasitic infection. After transfer-blot of recombinant mTsMP separated by 12% SDS-PAGE, each strip was incubated with the respective serum in a dilution of 1:200, subsequently with 1:1,000 diluted peroxidase conjugated anti-human IgG. The colour reactions were developed with 4-chloro-1-naphthol (4C1N) and 0.03% hydrogen peroxide (H2O2). Panel TsM, cysticercosis; panel Ec, echinococcosis; panel Sp, sparganosis; panel Pw, paragonimiasis; panel Cs, clonorchiasis; panel Normal, healthy controls. Lanes 1-10, each represent serum from different patient. The low molecular weight marker in kilodaltons (kDa) was shown at the left
Mentions: To assess the diagnostic value of recombinant mTsMP, we tested its immunoreactivity by an immunoblot assay using individual sera from patients with various helminthic infections including cysticercosis, echinococcosis, sparganosis, paragonimiasis and clonorchiasis. A typical result of the immunoblot analysis is shown in Fig. 5. A strong recognition of recombinant mTsMP was observed in 87.5% (35/40 cases) of sera from patients with cysticercosis who were examined (Fig. 5, panel TsM). Meanwhile, weak cross-reactivity was observed in only 1 sample from a patient with echinococcosis (Fig. 5, panel Ec). None of the sera from patients with other parasitic infections or from normal controls showed any antibody positive reactivity (Fig. 5, panel Sp, Pw, Cs and Normal).

Bottom Line: After refolding and purification, enzymatic properties of the recombinant metalloproteinase were observed.The recombinant TsMP protein showed proteolytic activity, which preferred host extracellular matrix proteins such as collagen and fibronectin as degradable substrates.TsMP might be involved in the processing of numerous host proteins and play an important role in the parasite life cycle.

View Article: PubMed Central - PubMed

Affiliation: Dept. of Medical Genetics, Wuhan University School of Basic Medicial Sciences, Wuhan 430071, China.

ABSTRACT

Background: Parasite proteases have important roles in cleavage of host proteins during the invasion of host tissues and participate in the parasite's evasion from the host's immune response. The aim of the present study was to estimate a metalloproteinase properties of Taenia solium metacestode (TsMP) during host-parasite interactions, and evaluate its potential as a serodiagnostic antigen for cysticercosis.

Methods: The cDNA coding for the mature catalytic domain of TsMP was cloned into pGEX-6P-1 expression vector. A recombinant glutathione S-transferase and TsMP fusion protein was induced. After refolding and purification, enzymatic properties of the recombinant metalloproteinase were observed. Immunoblot assay was processed to evaluate its potential as a serodiagnostic antigen for cysticercosis.

Results: The recombinant TsMP protein showed proteolytic activity, which preferred host extracellular matrix proteins such as collagen and fibronectin as degradable substrates. In immunoblot assay, 87.5% of sera from patients with cysticercosis showed strong reactivity. In sera from patients with other parasitic infections and from normal controls, it showed high specificity.

Conclusions: TsMP might be involved in the processing of numerous host proteins and play an important role in the parasite life cycle. A single recombinant TsMP antigen could have a potential value for serodiagnosis of cysticercosis.

No MeSH data available.


Related in: MedlinePlus