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Analyzing and identifying novel B cell epitopes within Toxoplasma gondii GRA4.

Wang Y, Wang G, Ou J, Yin H, Zhang D - Parasit Vectors (2014)

Bottom Line: The potential B cell epitopes of GRA4 predicted by bioinformatics tools focused on six regions of GRA4, 52-77 aa, 93-112 aa, 127-157 aa, 178-201 aa, 223-252 aa and 314-333 aa.Three of the eleven peptides (amino acids 62-77, 233-252 and 314-333) tested were recognized by all sera.The identified epitopes may be useful for additional studies of epitope-based vaccines and diagnostic reagents.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, 730046, China. wangyh061001@163.com.

ABSTRACT

Background: The identification of specific epitopes targeted by the host antibody response is important for understanding the natural response to infection and for the development of epitope-based marker vaccines and diagnostic tools for toxoplasmosis. In this study, Toxoplasma gondii GRA4 epitopes were identified using software-based prediction and a synthetic peptide technique.

Methods: The complete GRA4 gene sequence was obtained from T. gondii of the Gansu Jingtai strain of tachyzoites. The potential B cell epitopes of GRA4 was predicted using the PROTEAN subroutine in the DNASTAR software package. The peptides with good hydrophilicity, high accessibility, high flexibility and strong antigenicity were chemically synthesized and assessed by ELISA using pig sera from different time points after infection.

Results: The potential B cell epitopes of GRA4 predicted by bioinformatics tools focused on six regions of GRA4, 52-77 aa, 93-112 aa, 127-157 aa, 178-201 aa, 223-252 aa and 314-333 aa. Eleven shorter peptides from the six regions were synthesized and assessed by ELISA using pig sera from different time points after infection. Three of the eleven peptides (amino acids 62-77, 233-252 and 314-333) tested were recognized by all sera.

Conclusions: We precisely located the T. gondii GRA4 epitopes using pig sera collected at different time points after infection. The identified epitopes may be useful for additional studies of epitope-based vaccines and diagnostic reagents.

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Related in: MedlinePlus

ELISA of IgG antibodies against an irrelevant peptide (A), STAg (B) and recombinant GRA4 (C) in the four groups of pig sera. The cut-off point for the assay is indicated by the horizontal line.
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Fig4: ELISA of IgG antibodies against an irrelevant peptide (A), STAg (B) and recombinant GRA4 (C) in the four groups of pig sera. The cut-off point for the assay is indicated by the horizontal line.

Mentions: To determine the specificity of the anti-peptide antibody, an ELISA using an irrelevant peptide was also performed. This peptide did not react with the sera (Figure 4A). To compare the serological reactivity of the peptides with STAg and recombinant GRA4, ELISAs using STAg and recombinant GRA4 were also performed. A total of 51 sera samples reacted with STAg (Figure 4B) and recombinant GRA4 (Figure 4C).Figure 4


Analyzing and identifying novel B cell epitopes within Toxoplasma gondii GRA4.

Wang Y, Wang G, Ou J, Yin H, Zhang D - Parasit Vectors (2014)

ELISA of IgG antibodies against an irrelevant peptide (A), STAg (B) and recombinant GRA4 (C) in the four groups of pig sera. The cut-off point for the assay is indicated by the horizontal line.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4195951&req=5

Fig4: ELISA of IgG antibodies against an irrelevant peptide (A), STAg (B) and recombinant GRA4 (C) in the four groups of pig sera. The cut-off point for the assay is indicated by the horizontal line.
Mentions: To determine the specificity of the anti-peptide antibody, an ELISA using an irrelevant peptide was also performed. This peptide did not react with the sera (Figure 4A). To compare the serological reactivity of the peptides with STAg and recombinant GRA4, ELISAs using STAg and recombinant GRA4 were also performed. A total of 51 sera samples reacted with STAg (Figure 4B) and recombinant GRA4 (Figure 4C).Figure 4

Bottom Line: The potential B cell epitopes of GRA4 predicted by bioinformatics tools focused on six regions of GRA4, 52-77 aa, 93-112 aa, 127-157 aa, 178-201 aa, 223-252 aa and 314-333 aa.Three of the eleven peptides (amino acids 62-77, 233-252 and 314-333) tested were recognized by all sera.The identified epitopes may be useful for additional studies of epitope-based vaccines and diagnostic reagents.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, 730046, China. wangyh061001@163.com.

ABSTRACT

Background: The identification of specific epitopes targeted by the host antibody response is important for understanding the natural response to infection and for the development of epitope-based marker vaccines and diagnostic tools for toxoplasmosis. In this study, Toxoplasma gondii GRA4 epitopes were identified using software-based prediction and a synthetic peptide technique.

Methods: The complete GRA4 gene sequence was obtained from T. gondii of the Gansu Jingtai strain of tachyzoites. The potential B cell epitopes of GRA4 was predicted using the PROTEAN subroutine in the DNASTAR software package. The peptides with good hydrophilicity, high accessibility, high flexibility and strong antigenicity were chemically synthesized and assessed by ELISA using pig sera from different time points after infection.

Results: The potential B cell epitopes of GRA4 predicted by bioinformatics tools focused on six regions of GRA4, 52-77 aa, 93-112 aa, 127-157 aa, 178-201 aa, 223-252 aa and 314-333 aa. Eleven shorter peptides from the six regions were synthesized and assessed by ELISA using pig sera from different time points after infection. Three of the eleven peptides (amino acids 62-77, 233-252 and 314-333) tested were recognized by all sera.

Conclusions: We precisely located the T. gondii GRA4 epitopes using pig sera collected at different time points after infection. The identified epitopes may be useful for additional studies of epitope-based vaccines and diagnostic reagents.

Show MeSH
Related in: MedlinePlus