| Doctorant : |
Maruthi PRASANNA
|
| Directeur de thèse : |
Cyrille GRANDJEAN ,
Directeur de recherche CNRS |
| co-directeur de thèse : | Noemí Csaba, Associate Professor, University of Santiago de Compostela, Spain |
| Encadrant : |
Cyrille GRANDJEAN ,
Directeur de recherche CNRS |
| Financement : |
NANOFAR |
| Date de la soutenance : |
vendredi 11 septembre 2020, 11h00 |
| Modalité : |
|
| Jury : |
- Président de jury : Antonio Almeida, Professor, University of Lisbon
- Rapporteur : Maria J. Alonso Fernandez, Professor, Santiago de Compostela University
- Rapporteur : Dr. Rosana Simon, Lecturer, University of Vigo
- Examinateur :
Bernard OFFMANN ,
Professeur Université
- Directeur de thèse :
Cyrille GRANDJEAN ,
Directeur de recherche CNRS
- co-directeur de thèse : Noemí Csaba, Associate Professor, University of Santiago de Compostela, Spain
- Encadrant :
Cyrille GRANDJEAN ,
Directeur de recherche CNRS
|
This study is of great importance for the advancement in the development of pneumococcal vaccines and the improvement of their efficacy. Indeed, researchers have succeeded in preparing and characterizing chitosan nanoparticles loaded with antigen, suitable for immunization.
The currently available pneumococcal vaccines are not effective in complete prevention of pneumococcal infections. To develop a better vaccine, first, “we were able to produce a glycoconjugate antigen (molecule resulting from the binding of carbohydrates to proteins or carbohydrates to lipids) based on a common pneumococcal vaccine. On the basis of the positive immune responses generated by the glycoconjugate antigen in mice, we further envisaged that their combination with the nanotechnologies would generate a vaccine with enhanced potential,” explains Maruthi Prasanna.
Following this approach, chitosan nanoparticles were used to encapsulate the glycoconjugate. Finally, the developed nanovaccines generated an enhanced immune response against both the protein and polysaccharide components when compared to the naked glycoconjugate.
Publications
2019
Pillot, Aline; Defontaine, Alain; Fateh, Amina; Lambert, Annie; Prasanna, Maruthi; Fanuel, Mathieu; Pipelier, Muriel; Csaba, Noemi; Violo, Typhaine; Camberlein, Emilie; Grandjean, Cyrille
Site-Specific Conjugation for Fully Controlled Glycoconjugate Vaccine Preparation Article de journal
Dans: Frontiers in Chemistry, vol. 7, no. November, p. 1–9, 2019, ISSN: 22962646.
@article{Pillot2019,
title = {Site-Specific Conjugation for Fully Controlled Glycoconjugate Vaccine Preparation},
author = {Aline Pillot and Alain Defontaine and Amina Fateh and Annie Lambert and Maruthi Prasanna and Mathieu Fanuel and Muriel Pipelier and Noemi Csaba and Typhaine Violo and Emilie Camberlein and Cyrille Grandjean},
doi = {10.3389/fchem.2019.00726},
issn = {22962646},
year = {2019},
date = {2019-01-01},
journal = {Frontiers in Chemistry},
volume = {7},
number = {November},
pages = {1--9},
abstract = {Glycoconjugate vaccines are formed by covalently link a carbohydrate antigen to a carrier protein whose role is to achieve a long lasting immune response directed against the carbohydrate antigen. The nature of the sugar antigen, its length, its ratio per carrier protein and the conjugation chemistry impact on both structure and the immune response of a glycoconjugate vaccine. In addition it has long been assumed that the sites at which the carbohydrate antigen is attached can also have an impact. These important issue can now be addressed owing to the development of novel chemoselective ligation reactions as well as techniques such as site-selective mutagenesis, glycoengineering, or extension of the genetic code. The preparation and characterization of homogeneous bivalent pneumococcal vaccines is reported. The preparation and characterization of homogeneous bivalent pneumococcal vaccines is reported. A synthetic tetrasaccharide representative of the serotype 14 capsular polysaccharide of Streptococcus pneumoniae has been linked using the thiol/maleimide coupling chemistry to four different Pneumococcal surface adhesin A (PsaA) mutants, each harboring a single cysteine mutation at a defined position. Humoral response of these 1 to 1 carbohydrate antigen/PsaA conjugates have been assessed in mice. Our results showed that the carbohydrate antigen-PsaA connectivity impacts the anti-carrier response and raise questions about the design of glycoconjugate vaccine whereby the protein plays the dual role of immunogen and carrier.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Glycoconjugate vaccines are formed by covalently link a carbohydrate antigen to a carrier protein whose role is to achieve a long lasting immune response directed against the carbohydrate antigen. The nature of the sugar antigen, its length, its ratio per carrier protein and the conjugation chemistry impact on both structure and the immune response of a glycoconjugate vaccine. In addition it has long been assumed that the sites at which the carbohydrate antigen is attached can also have an impact. These important issue can now be addressed owing to the development of novel chemoselective ligation reactions as well as techniques such as site-selective mutagenesis, glycoengineering, or extension of the genetic code. The preparation and characterization of homogeneous bivalent pneumococcal vaccines is reported. The preparation and characterization of homogeneous bivalent pneumococcal vaccines is reported. A synthetic tetrasaccharide representative of the serotype 14 capsular polysaccharide of Streptococcus pneumoniae has been linked using the thiol/maleimide coupling chemistry to four different Pneumococcal surface adhesin A (PsaA) mutants, each harboring a single cysteine mutation at a defined position. Humoral response of these 1 to 1 carbohydrate antigen/PsaA conjugates have been assessed in mice. Our results showed that the carbohydrate antigen-PsaA connectivity impacts the anti-carrier response and raise questions about the design of glycoconjugate vaccine whereby the protein plays the dual role of immunogen and carrier.
Prasanna, Maruthi; Soulard, Daphnée; Camberlein, Emilie; Ruffier, Nicolas; Lambert, Annie; Trottein, François; Csaba, Noemi; Grandjean, Cyrille
Semisynthetic glycoconjugate based on dual role protein/PsaA as a pneumococcal vaccine Article de journal
Dans: European Journal of Pharmaceutical Sciences, vol. 129, p. 31–41, 2019, ISSN: 0928-0987.
@article{PRASANNA201931,
title = {Semisynthetic glycoconjugate based on dual role protein/PsaA as a pneumococcal vaccine},
author = {Maruthi Prasanna and Daphnée Soulard and Emilie Camberlein and Nicolas Ruffier and Annie Lambert and François Trottein and Noemi Csaba and Cyrille Grandjean},
url = {http://www.sciencedirect.com/science/article/pii/S0928098718305487},
doi = {https://doi.org/10.1016/j.ejps.2018.12.013},
issn = {0928-0987},
year = {2019},
date = {2019-01-01},
journal = {European Journal of Pharmaceutical Sciences},
volume = {129},
pages = {31--41},
abstract = {Pneumococcal infections remain a major public health concern worldwide. The currently available vaccines in the market are based on pneumococcal capsular polysaccharides but they still need to be improved to secure an optimal coverage notably in population at risk. To circumvent this, association of virulence pneumococcal proteins to the polysaccharide valencies has been proposed with the hope to observe an additive - if not synergistic - protective effect. Along this line, the use of the highly conserved and ubiquitous pneumococcal surface adhesin A (PsaA) as a protein carrier for a synthetic pneumococcal oligosaccharide is demonstrated herein for the first time. A tetrasaccharide mimicking functional antigenic determinants from the S. pneumoniae serotype 14 capsular polysaccharide (Pn14TS) was chemically synthesised. The mature PsaA (mPsaA) was expressed in E. coli and purified using affinity chromatography. The Pn14PS was conjugated to mPsaA using maleimide-thiol coupling chemistry to obtain mPsaA-Pn14PS conjugate (protein/sugar molar ratio: 1/5.4). The mPsaA retained the structural conformation after the conjugation and lyophilisation. The prepared glycoconjugate adjuvanted with α-galactosylceramide, a potent activator of invariant Natural Killer T cells, was tested in mice for its immunological response upon subcutaneous injection in comparison with mPsaA alone and a model BSA conjugate (BSA-Pn14PS, used here as a control). Mice immunised with the mPsaA-Pn14TS produced a robust IgG response against mPsaA and against the capsular polysaccharide from pneumococcal serotype 14. These data provide the basis for novel pneumococcal vaccine development.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Pneumococcal infections remain a major public health concern worldwide. The currently available vaccines in the market are based on pneumococcal capsular polysaccharides but they still need to be improved to secure an optimal coverage notably in population at risk. To circumvent this, association of virulence pneumococcal proteins to the polysaccharide valencies has been proposed with the hope to observe an additive - if not synergistic - protective effect. Along this line, the use of the highly conserved and ubiquitous pneumococcal surface adhesin A (PsaA) as a protein carrier for a synthetic pneumococcal oligosaccharide is demonstrated herein for the first time. A tetrasaccharide mimicking functional antigenic determinants from the S. pneumoniae serotype 14 capsular polysaccharide (Pn14TS) was chemically synthesised. The mature PsaA (mPsaA) was expressed in E. coli and purified using affinity chromatography. The Pn14PS was conjugated to mPsaA using maleimide-thiol coupling chemistry to obtain mPsaA-Pn14PS conjugate (protein/sugar molar ratio: 1/5.4). The mPsaA retained the structural conformation after the conjugation and lyophilisation. The prepared glycoconjugate adjuvanted with α-galactosylceramide, a potent activator of invariant Natural Killer T cells, was tested in mice for its immunological response upon subcutaneous injection in comparison with mPsaA alone and a model BSA conjugate (BSA-Pn14PS, used here as a control). Mice immunised with the mPsaA-Pn14TS produced a robust IgG response against mPsaA and against the capsular polysaccharide from pneumococcal serotype 14. These data provide the basis for novel pneumococcal vaccine development.
Lien
