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Vaccination against Streptococcus pneumoniae using truncated derivatives of polyhistidine triad protein D.

Plumptre CD, Ogunniyi AD, Paton JC - PLoS ONE (2013)

Bottom Line: In this study, we purified recombinant truncated derivatives of PhtD and examined their secondary structural composition, as well as their capacity to bind antibodies from polyclonal murine serum generated against the full length protein.This allowed the identification of a particularly immunogenic fragment of PhtD, which was also purified and characterised.These experiments revealed that whilst the immunogenic region identified may be a promising candidate to protect against sepsis, the full length PhtD was ineffective at conferring significant protective immunity.

View Article: PubMed Central - PubMed

Affiliation: Research Centre for Infectious Diseases, School of Molecular and Biomedical Science, University of Adelaide, Adelaide, South Australia, Australia.

ABSTRACT
Polyhistidine triad protein D (PhtD) has been described as a promising vaccine candidate for use against Streptococcus pneumoniae, but there has been a lack of examination of its structure and of which region(s) of the protein are targeted by protective immune responses. In this study, we purified recombinant truncated derivatives of PhtD and examined their secondary structural composition, as well as their capacity to bind antibodies from polyclonal murine serum generated against the full length protein. This allowed the identification of a particularly immunogenic fragment of PhtD, which was also purified and characterised. The truncated derivatives were tested as vaccine antigens in mouse models of pneumococcal sepsis and colonisation, using alum and E. coli heat labile toxin B subunit respectively as adjuvants. These experiments revealed that whilst the immunogenic region identified may be a promising candidate to protect against sepsis, the full length PhtD was ineffective at conferring significant protective immunity. These results are significant for the potential for PhtD to be used in novel vaccines, which are currently being tested in clinical trials.

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Related in: MedlinePlus

SDS-PAGE analysis of purified truncated derivatives of PhtD.Truncated derivatives of PhtD were analysed by SDS-PAGE using a 12% acrylamide gel and subsequently stained using Coomassie R250. 1 µg of each protein was used. The identity of the protein is indicated above each lane (FL; full length).
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pone-0078916-g003: SDS-PAGE analysis of purified truncated derivatives of PhtD.Truncated derivatives of PhtD were analysed by SDS-PAGE using a 12% acrylamide gel and subsequently stained using Coomassie R250. 1 µg of each protein was used. The identity of the protein is indicated above each lane (FL; full length).

Mentions: The protein products of the cloned genes were expressed in E. coli and purified by metal affinity chromatography. All were judged to be >90% pure by Coomassie staining following SDS-PAGE analysis (see Figure 3).


Vaccination against Streptococcus pneumoniae using truncated derivatives of polyhistidine triad protein D.

Plumptre CD, Ogunniyi AD, Paton JC - PLoS ONE (2013)

SDS-PAGE analysis of purified truncated derivatives of PhtD.Truncated derivatives of PhtD were analysed by SDS-PAGE using a 12% acrylamide gel and subsequently stained using Coomassie R250. 1 µg of each protein was used. The identity of the protein is indicated above each lane (FL; full length).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3814962&req=5

pone-0078916-g003: SDS-PAGE analysis of purified truncated derivatives of PhtD.Truncated derivatives of PhtD were analysed by SDS-PAGE using a 12% acrylamide gel and subsequently stained using Coomassie R250. 1 µg of each protein was used. The identity of the protein is indicated above each lane (FL; full length).
Mentions: The protein products of the cloned genes were expressed in E. coli and purified by metal affinity chromatography. All were judged to be >90% pure by Coomassie staining following SDS-PAGE analysis (see Figure 3).

Bottom Line: In this study, we purified recombinant truncated derivatives of PhtD and examined their secondary structural composition, as well as their capacity to bind antibodies from polyclonal murine serum generated against the full length protein.This allowed the identification of a particularly immunogenic fragment of PhtD, which was also purified and characterised.These experiments revealed that whilst the immunogenic region identified may be a promising candidate to protect against sepsis, the full length PhtD was ineffective at conferring significant protective immunity.

View Article: PubMed Central - PubMed

Affiliation: Research Centre for Infectious Diseases, School of Molecular and Biomedical Science, University of Adelaide, Adelaide, South Australia, Australia.

ABSTRACT
Polyhistidine triad protein D (PhtD) has been described as a promising vaccine candidate for use against Streptococcus pneumoniae, but there has been a lack of examination of its structure and of which region(s) of the protein are targeted by protective immune responses. In this study, we purified recombinant truncated derivatives of PhtD and examined their secondary structural composition, as well as their capacity to bind antibodies from polyclonal murine serum generated against the full length protein. This allowed the identification of a particularly immunogenic fragment of PhtD, which was also purified and characterised. The truncated derivatives were tested as vaccine antigens in mouse models of pneumococcal sepsis and colonisation, using alum and E. coli heat labile toxin B subunit respectively as adjuvants. These experiments revealed that whilst the immunogenic region identified may be a promising candidate to protect against sepsis, the full length PhtD was ineffective at conferring significant protective immunity. These results are significant for the potential for PhtD to be used in novel vaccines, which are currently being tested in clinical trials.

Show MeSH
Related in: MedlinePlus