High antibody titres induced by protein subunit vaccines using Mycobacterium ulcerans antigens Hsp18 and MUL_3720 with a TLR-2 agonist fail to protect against Buruli ulcer in mice

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Biochemistry, Biophysics and Molecular Biology

Main article text

 

Introduction

Materials & Methods

Strains and culture conditions

Recombinant protein expression

Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE)

Western Blotting

Analysis of electrostatic interaction between protein antigen and lipopeptide formulations

Lipopeptide vaccine preparation

Ethics statement for animal experiments

Vaccination of animals

M. ulcerans challenge

Serum antibody titre measurements

Statistical analysis

Results

Recombinant MUL_3720 and Hsp18 both bound to R4Pam2Cys

Vaccination induced strong protein-specific antibody responses

Measurement of IgG antibody subtypes following MUL_3720 + R4Pam2Cys and Hsp18 + R4Pam2Cys vaccination

MUL_3720 + R4Pam2Cys and Hsp18 + R4Pam2Cys do not protect against the onset of BU

Antibody titres do not correlate with protection against M. ulcerans

Challenge with M. ulcerans did not induce protein-specific antibody levels comparable to vaccination with MUL_3720 or Hsp18

Discussion

Conclusions

Supplemental Information

Summary of protein antigens and assay characteristics

DOI: 10.7717/peerj.9659/supp-1

Raw data for protein-adjuvant binding

DOI: 10.7717/peerj.9659/supp-2

Antibody titre raw data

DOI: 10.7717/peerj.9659/supp-3

Survival outcomes BALB/C mice

DOI: 10.7717/peerj.9659/supp-4

S urvival outcomes in vaccinated C57B6 mice

DOI: 10.7717/peerj.9659/supp-5

Additional Information and Declarations

Competing Interests

Timothy Stinear is an Academic Editor for PeerJ.

Author Contributions

Kirstie M. Mangas, Nicholas J. Tobias, Estelle Marion and Laurent Marsollier conceived and designed the experiments, performed the experiments, analyzed the data, prepared figures and/or tables, authored or reviewed drafts of the paper, and approved the final draft.

Jérémie Babonneau performed the experiments, analyzed the data, prepared figures and/or tables, authored or reviewed drafts of the paper, and approved the final draft.

Jessica L. Porter performed the experiments, authored or reviewed drafts of the paper, and approved the final draft.

Sacha J. Pidot and Timothy P. Stinear conceived and designed the experiments, analyzed the data, prepared figures and/or tables, authored or reviewed drafts of the paper, and approved the final draft.

Chinn Yi Wong performed the experiments, prepared figures and/or tables, authored or reviewed drafts of the paper, and approved the final draft.

David C. Jackson conceived and designed the experiments, analyzed the data, authored or reviewed drafts of the paper, and approved the final draft.

Brendon Y. Chua conceived and designed the experiments, performed the experiments, analyzed the data, prepared figures and/or tables, authored or reviewed drafts of the paper, and approved the final draft.

Animal Ethics

The following information was supplied relating to ethical approvals (i.e., approving body and any reference numbers):

The Ethics Committee of region Pays de la Loire provided full approval for this research under protocol nos. CEEA 2009.14 and CEEA 2012.145.

Data Availability

The following information was supplied regarding data availability:

The raw measurements are available as a Supplemental File.

Funding

This work was supported by the National Health and Medical Research Council of Australia (GNT1105525). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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