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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

Recognition of predicted epitope peptides by rabbit anti-FliCBp sera as detected by Indirect ELISA.Controls (A) composed of human serum albumin (HSA), crude B. pseudomallei, recombinant FliCBp, without primary antibody (No 10 Ab) and without secondary antibody (No 20 Ab) and F51-69, F96-111, F270-288 and F213-231 peptides (B) were coated at various concentration (1, 3 and 10 μg/ml) onto a 96-well polystyrene plate and probed with diluted rabbit antibodies to FliCBp and quantified by indirect ELISA. Results are represented by the Absorbance index (O.D.test−O.D.uncoated / O.D.uncoated). Experiments were performed in duplicate and results represent the mean of the Absorbance index ± SE.
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pntd.0003917.g002: Recognition of predicted epitope peptides by rabbit anti-FliCBp sera as detected by Indirect ELISA.Controls (A) composed of human serum albumin (HSA), crude B. pseudomallei, recombinant FliCBp, without primary antibody (No 10 Ab) and without secondary antibody (No 20 Ab) and F51-69, F96-111, F270-288 and F213-231 peptides (B) were coated at various concentration (1, 3 and 10 μg/ml) onto a 96-well polystyrene plate and probed with diluted rabbit antibodies to FliCBp and quantified by indirect ELISA. Results are represented by the Absorbance index (O.D.test−O.D.uncoated / O.D.uncoated). Experiments were performed in duplicate and results represent the mean of the Absorbance index ± SE.

Mentions: Predicted B and T cell epitopes were initially evaluated for immunorecognition by probing them with rabbit anti-FliCBp anti-sera. All three peptides were recognized (Fig 2). Subsequently, crude B. pseudomallei extract (Bps), recombinant FliCBp, and peptides F51-69, F96-111 and F270-288 were tested against human plasma samples for immunorecognition (Fig 3). In order to investigate a possible role of the peptides in protection, samples were tested from diverse sub-populations, including healthy seronegative, healthy seropositive and recovered melioidosis patient groups. Results showed that F51-69 and F270-288 were recognized by antibodies in human plasma, in contrast to F96-111. In addition, antisera from recovered individuals were shown to recognize Bps extract, FliCBp, and all peptides, except for F96-111, to a greater extent than those from healthy seropositive and seronegative individuals (Fig 3).


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)

Recognition of predicted epitope peptides by rabbit anti-FliCBp sera as detected by Indirect ELISA.Controls (A) composed of human serum albumin (HSA), crude B. pseudomallei, recombinant FliCBp, without primary antibody (No 10 Ab) and without secondary antibody (No 20 Ab) and F51-69, F96-111, F270-288 and F213-231 peptides (B) were coated at various concentration (1, 3 and 10 μg/ml) onto a 96-well polystyrene plate and probed with diluted rabbit antibodies to FliCBp and quantified by indirect ELISA. Results are represented by the Absorbance index (O.D.test−O.D.uncoated / O.D.uncoated). Experiments were performed in duplicate and results represent the mean of the Absorbance index ± SE.
© Copyright Policy
Related In: Results  -  Collection

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

pntd.0003917.g002: Recognition of predicted epitope peptides by rabbit anti-FliCBp sera as detected by Indirect ELISA.Controls (A) composed of human serum albumin (HSA), crude B. pseudomallei, recombinant FliCBp, without primary antibody (No 10 Ab) and without secondary antibody (No 20 Ab) and F51-69, F96-111, F270-288 and F213-231 peptides (B) were coated at various concentration (1, 3 and 10 μg/ml) onto a 96-well polystyrene plate and probed with diluted rabbit antibodies to FliCBp and quantified by indirect ELISA. Results are represented by the Absorbance index (O.D.test−O.D.uncoated / O.D.uncoated). Experiments were performed in duplicate and results represent the mean of the Absorbance index ± SE.
Mentions: Predicted B and T cell epitopes were initially evaluated for immunorecognition by probing them with rabbit anti-FliCBp anti-sera. All three peptides were recognized (Fig 2). Subsequently, crude B. pseudomallei extract (Bps), recombinant FliCBp, and peptides F51-69, F96-111 and F270-288 were tested against human plasma samples for immunorecognition (Fig 3). In order to investigate a possible role of the peptides in protection, samples were tested from diverse sub-populations, including healthy seronegative, healthy seropositive and recovered melioidosis patient groups. Results showed that F51-69 and F270-288 were recognized by antibodies in human plasma, in contrast to F96-111. In addition, antisera from recovered individuals were shown to recognize Bps extract, FliCBp, and all peptides, except for F96-111, to a greater extent than those from healthy seropositive and seronegative individuals (Fig 3).

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