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Sequence- and Structure-Based Immunoreactive Epitope Discovery for Burkholderia pseudomallei Flagellin.

Nithichanon A, Rinchai D, Gori A, Lassaux P, Peri C, Conchillio-Solé O, Ferrer-Navarro M, Gourlay LJ, Nardini M, Vila J, Daura X, Colombo G, Bolognesi M, Lertmemonkolchai G - PLoS Negl Trop Dis (2015)

Bottom Line: Due to its inherent resistance to the major antibiotic classes and its facultative intracellular pathogenicity, an effective vaccine would be extremely desirable, along with appropriate prevention and therapeutic management.All three epitopes were shown to be immunoreactive against human IgG antibodies and to elicit cytokine production from human peripheral blood mononuclear cells.Furthermore, two of the peptides (F51-69 and F270-288) were found to be dominant immunoreactive epitopes, and their antibodies enhanced the bactericidal activities of purified human neutrophils.

View Article: PubMed Central - PubMed

Affiliation: The Centre for Research and Development of Medical Diagnostic Laboratories, Faculty of Associated Medical Sciences, Khon Kaen University, Khon Kaen, Thailand.

ABSTRACT
Burkholderia pseudomallei is a Gram-negative bacterium responsible for melioidosis, a serious and often fatal infectious disease that is poorly controlled by existing treatments. Due to its inherent resistance to the major antibiotic classes and its facultative intracellular pathogenicity, an effective vaccine would be extremely desirable, along with appropriate prevention and therapeutic management. One of the main subunit vaccine candidates is flagellin of Burkholderia pseudomallei (FliCBp). Here, we present the high resolution crystal structure of FliCBp and report the synthesis and characterization of three peptides predicted to be both B and T cell FliCBp epitopes, by both structure-based in silico methods, and sequence-based epitope prediction tools. All three epitopes were shown to be immunoreactive against human IgG antibodies and to elicit cytokine production from human peripheral blood mononuclear cells. Furthermore, two of the peptides (F51-69 and F270-288) were found to be dominant immunoreactive epitopes, and their antibodies enhanced the bactericidal activities of purified human neutrophils. The epitopes derived from this study may represent potential melioidosis vaccine components.

No MeSH data available.


Related in: MedlinePlus

FliCBp B-cell linear sequence-based epitope predictions using three predictor methods: BepiPred (threshold = 0.35), BCPred (classifier specificity = 75%), and AAP (classifier specificity = 75%).Epitope predictions were carried out on the full-length amino acid sequence of FliCBp, using three web-accessible prediction servers: BepiPred, BCPred and AAP [21,22]. Predicted epitope residues by BepiPred, BCPred and AAP are shown in red, blue and green font, respectively on the amino acid sequence (residue numbers are indicated). Grey shaded boxes indicate consensus positive residues identified by at least two epitope predictors.
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pntd.0003917.g001: FliCBp B-cell linear sequence-based epitope predictions using three predictor methods: BepiPred (threshold = 0.35), BCPred (classifier specificity = 75%), and AAP (classifier specificity = 75%).Epitope predictions were carried out on the full-length amino acid sequence of FliCBp, using three web-accessible prediction servers: BepiPred, BCPred and AAP [21,22]. Predicted epitope residues by BepiPred, BCPred and AAP are shown in red, blue and green font, respectively on the amino acid sequence (residue numbers are indicated). Grey shaded boxes indicate consensus positive residues identified by at least two epitope predictors.

Mentions: Epitope predictions were carried out on the full-length (residues 1 to 388) amino acid sequence of FliCBp (UNIPROT code H7C7G3), using the web-accessible prediction servers BepiPred, BCPred and AAP [21,22]. Six linear B cell epitopes were identified (Fig 1) and subsequently used for T cell epitope predictions using the IEDB server [23,48] (www.iedb.org), selecting for HLA DRB1 alleles common in Thailand; HLA DRB1*0301, 0405, 07, 09, 1202, 1501, 1502 and 1602 [28]. A detailed overview of the consensus between linear epitopes predicted by sequence-based web servers is shown in Fig 1. The top three epitopes that were predicted to be both B and T cell epitopes are F51-69, F96-111 and F270-288. Interestingly, F51-69 overlaps with Peptide 6 (residues 51–70); F96-111 overlaps with Peptide 10 (residues 91–110), and F270-288 overlaps with Peptide 28 (residues 271–290) from the peptide panel synthesized by Musson et al. The following peptides were synthesized as described in the Materials and Methods section: F51-69; 51-TRMQTQINGLNQGVSNAND-69, F96-111; 96-VQASNGPLSASDASAL-111, F270-288; 270-NATAMVAQINAVNKPQTVS-288.


Sequence- and Structure-Based Immunoreactive Epitope Discovery for Burkholderia pseudomallei Flagellin.

Nithichanon A, Rinchai D, Gori A, Lassaux P, Peri C, Conchillio-Solé O, Ferrer-Navarro M, Gourlay LJ, Nardini M, Vila J, Daura X, Colombo G, Bolognesi M, Lertmemonkolchai G - PLoS Negl Trop Dis (2015)

FliCBp B-cell linear sequence-based epitope predictions using three predictor methods: BepiPred (threshold = 0.35), BCPred (classifier specificity = 75%), and AAP (classifier specificity = 75%).Epitope predictions were carried out on the full-length amino acid sequence of FliCBp, using three web-accessible prediction servers: BepiPred, BCPred and AAP [21,22]. Predicted epitope residues by BepiPred, BCPred and AAP are shown in red, blue and green font, respectively on the amino acid sequence (residue numbers are indicated). Grey shaded boxes indicate consensus positive residues identified by at least two epitope predictors.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4519301&req=5

pntd.0003917.g001: FliCBp B-cell linear sequence-based epitope predictions using three predictor methods: BepiPred (threshold = 0.35), BCPred (classifier specificity = 75%), and AAP (classifier specificity = 75%).Epitope predictions were carried out on the full-length amino acid sequence of FliCBp, using three web-accessible prediction servers: BepiPred, BCPred and AAP [21,22]. Predicted epitope residues by BepiPred, BCPred and AAP are shown in red, blue and green font, respectively on the amino acid sequence (residue numbers are indicated). Grey shaded boxes indicate consensus positive residues identified by at least two epitope predictors.
Mentions: Epitope predictions were carried out on the full-length (residues 1 to 388) amino acid sequence of FliCBp (UNIPROT code H7C7G3), using the web-accessible prediction servers BepiPred, BCPred and AAP [21,22]. Six linear B cell epitopes were identified (Fig 1) and subsequently used for T cell epitope predictions using the IEDB server [23,48] (www.iedb.org), selecting for HLA DRB1 alleles common in Thailand; HLA DRB1*0301, 0405, 07, 09, 1202, 1501, 1502 and 1602 [28]. A detailed overview of the consensus between linear epitopes predicted by sequence-based web servers is shown in Fig 1. The top three epitopes that were predicted to be both B and T cell epitopes are F51-69, F96-111 and F270-288. Interestingly, F51-69 overlaps with Peptide 6 (residues 51–70); F96-111 overlaps with Peptide 10 (residues 91–110), and F270-288 overlaps with Peptide 28 (residues 271–290) from the peptide panel synthesized by Musson et al. The following peptides were synthesized as described in the Materials and Methods section: F51-69; 51-TRMQTQINGLNQGVSNAND-69, F96-111; 96-VQASNGPLSASDASAL-111, F270-288; 270-NATAMVAQINAVNKPQTVS-288.

Bottom Line: Due to its inherent resistance to the major antibiotic classes and its facultative intracellular pathogenicity, an effective vaccine would be extremely desirable, along with appropriate prevention and therapeutic management.All three epitopes were shown to be immunoreactive against human IgG antibodies and to elicit cytokine production from human peripheral blood mononuclear cells.Furthermore, two of the peptides (F51-69 and F270-288) were found to be dominant immunoreactive epitopes, and their antibodies enhanced the bactericidal activities of purified human neutrophils.

View Article: PubMed Central - PubMed

Affiliation: The Centre for Research and Development of Medical Diagnostic Laboratories, Faculty of Associated Medical Sciences, Khon Kaen University, Khon Kaen, Thailand.

ABSTRACT
Burkholderia pseudomallei is a Gram-negative bacterium responsible for melioidosis, a serious and often fatal infectious disease that is poorly controlled by existing treatments. Due to its inherent resistance to the major antibiotic classes and its facultative intracellular pathogenicity, an effective vaccine would be extremely desirable, along with appropriate prevention and therapeutic management. One of the main subunit vaccine candidates is flagellin of Burkholderia pseudomallei (FliCBp). Here, we present the high resolution crystal structure of FliCBp and report the synthesis and characterization of three peptides predicted to be both B and T cell FliCBp epitopes, by both structure-based in silico methods, and sequence-based epitope prediction tools. All three epitopes were shown to be immunoreactive against human IgG antibodies and to elicit cytokine production from human peripheral blood mononuclear cells. Furthermore, two of the peptides (F51-69 and F270-288) were found to be dominant immunoreactive epitopes, and their antibodies enhanced the bactericidal activities of purified human neutrophils. The epitopes derived from this study may represent potential melioidosis vaccine components.

No MeSH data available.


Related in: MedlinePlus