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Generation of chimeric antigen receptor (CAR) to recognize Paracoccidioides brasiliensis

Grant number: 22/13543-1
Support Opportunities:Scholarships in Brazil - Scientific Initiation
Effective date (Start): November 01, 2022
Effective date (End): October 31, 2023
Field of knowledge:Biological Sciences - Immunology - Applied Immunology
Principal Investigator:Thiago Aparecido da Silva
Grantee:Letícia Gregolato Alves
Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
Associated research grant:18/18538-0 - Bioengineered of T- and NK-cells by CAR against invasive fungal infections, AP.JP


Paracoccidioides brasiliensis is a pathogenic fungus belonging to the genus Paracoccidioides that has a restricted location a Latin America with the highest occurrence of cases in Brazil, Colombia and Venezuela. It is dimorphic and presents the carbohydrate ±-1,3-glucan, a virulence factor that compromises the recognition of the fungus by cells of innate immunity through pattern recognition receptors (PRRs). In addition to PPRs, CD4+ T cells with profiles Th1 and Th17 promote a protective antifungal response, however, the microenvironment at the infection site has a regulatory character and may prevent the recruitment of these effector immune cells. Based on this assumption, the current project proposes the use of the antigenic receptor (CAR) in the redirection of T cells to the site of infection to favor the direct action of cytotoxic cells on the fungus. Then, the current proposal aim to generate a CAR receptor constructs with specificity to P. brasiliensis. The cell wall components are important to redirect T cells to the infection site too and for that the target recognition domain present in the receptor CAR will be generated by the Phage Display technique, which will result in the identification of different scFv fragments. The project aims to evaluate the following parameters: (i) use of P. brasiliensis cell wall components for the murine model immunization and scFv coding sequence library construction; (ii) expression of the scFv library by the Phage Display technique to select scFv specific to the cell wall of P. brasiliensis; (iii) phage sequencing expressing scFv specific for P. brasiliensis; (iv) assembly of the constructs of the CAR receptors specific for P. brasiliensis in lentiviral vector with the object to express the constituent portions of the receptor.

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