Objective
This programme aims at the improvement of antigenic peptide-based vaccines by vehiculating these sequences through recombinant carrier proteins adequately tailored to this purpose.
Vaccine formulations, although available for a variety of diseases caused by infectious pathogens, can still be improved in a number of ways, such as the elimination of potential risks related to the use of live or inactivated vaccines. The use of subunit vaccines consisting of antigenic proteins or peptides derived from the pathogens responds to criteria of safety and, due to the exploitation of recombinant DNA technology or chemical synthesis, of reproducibility and economy. Several peptide vaccines have been proposed, but their general applicability has been hampered, among the others, by stability and toxicity problems exhibited by the synthetic molecules.
We propose here to use a combined approach derived both from the use as vaccines of peptide-based formulations and of recombinant proteins. We plan to construct, by insertion of appropriate oligonucleotides into suitable vectors, fusion proteins carrying peptides representing T- and B-cell epitopes in order to use them as possible vaccines. This technique should improve the stability and reduce the clearance of the antigenic peptides, possibly allowing to control its toxicity. This methodology also implies a greater reproducibility in comparison to the production of chemical adducts between synthetic peptides and carrier proteins. In recent years a growing interest in the study of peptide antigenicity in relation to the role of flanking sequences and protein topology in processing, presentation and recognition has been observed. The information available on the antigenicity of recombinant fusion proteins and their effect on the selection of antigen receptor repertoires is however limited. We thus plan to perform a systematic study of the ability of peptides representing B- and T-cell epitopes derived from HIV-l polypeptides to be recognized and to stimulate an immune response when inserted into different carrier molecules upon cloning of their coding sequences into suitable vectors and expression. We shall also try to include in antigenic recombinant proteins tagging sequences that could control their uptake by different presenting cells (APC) or address them into different processing compartments of the same APC. Furthermore, the limitations stemming from the genetic restrictions imposed by MHC haplotypes on T-cell recognition of the antigenic peptides to be presented to the immune system will be considered. To this end the interaction of peptides with MHC molecules will be analysed with the aim of designing peptides carrying agretopes (residues recognized by MHC antigens) allowing presentation by different MHC alleles without affecting the epitopes (residues recognized by T cell receptors). The work plan proposed thus includes: i) the choice, manipulation and functional characterization of different carrier proteins (monomeric or multimeric fusion proteins, multiple phage display vectors) engineered with T- and B-cell epitopes ii) the introduction and the study of the effect of multiple determinants in chimeric constructs including combinations of T- and B-epitopes or antigenic epitopes plus targeting determinants such as receptor binding peptides and sequences affecting protein internalization in a cell-specific or compartment-specific fashion iii) the systematic substitution of the residues of peptides to be inserted at appropriate positions of the carriers with the aim of abolishing the limitations imposed by haplotype restrictions on peptide/MHC interactions and thus on presentation to T-cells and stimulation of an effective immune response.
The laboratories involved in this project collectively display the know-how required to implement this programme. In particular, DNA recombinant techniques, methods for T-cell cloning and immunological analysis, procedures for phage display vectors construction and characterization, methodologies for the study of epitope conformations in chimeric molecules and for the molecular analysis of peptide-MHC interactions in living cells are all available to us. The participation of SANOFI as an associated partner, indicates that the research proposed can have at a later stage practical applications for the formulation of vaccines directed against specific diseases.
Fields of science (EuroSciVoc)
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques.
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques.
- natural sciencesbiological sciencesmicrobiologyvirology
- natural sciencesbiological sciencesgeneticsDNA
- natural sciencesbiological sciencesbiochemistrybiomoleculesproteins
- medical and health sciencesbasic medicineimmunology
- medical and health sciencesbasic medicinepharmacology and pharmacypharmaceutical drugsvaccines
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Topic(s)
Call for proposal
Data not availableFunding Scheme
CSC - Cost-sharing contractsCoordinator
94801 Villejuif
France