| Full text | |
| Author(s): Show less - |
Machado, Michele Procopio
;
dos Santos, Matheus Henrique
;
Guimaraes, Julia Garcia
;
de Campos, Gabriela Yamazaki
;
Oliveira Brito, Patricia Kellen Martins
;
Ferreira, Camilly Melo Garcia
;
Rezende, Caroline Patini
;
Frota, Natalia Fernandes
;
Soares, Sandro Gomes
;
Kumaresan, Pappanaicken R.
;
Lourenzoni, Marcos Roberto
;
da Silva, Thiago Aparecido
Total Authors: 12
|
| Document type: | Journal article |
| Source: | BIOENGINEERED; v. 14, n. 1, p. 26-pg., 2023-12-31. |
| Abstract | |
Cryptococcus spp. has a polysaccharide capsule composed of glucuronoxylomannan-GXM, a major virulence factor that can prevent the recognition of fungi by immune cells. Chimeric Antigen Receptor (CAR) redirects T cells to target Cryptococcus spp. as previously demonstrated by a CAR specific to GXM, GXMR-CAR. The current study evaluated the strength of the signal transduction triggered by GXMR-CAR, composed of a distinct antigen-binding domain sourced from a single-chain variable fragment (scFv). GXM-specific scFv derived from mAbs 2H1 and 18B7, 2H1-GXMR-CAR and 18B7-GXMR-CAR, respectively, were designed to express CD8 molecule as hinge/transmembrane, and the costimulatory molecule CD137 (4-1BB) coupled to CD3 zeta. The 2H1-GXMR-CAR or 18B7-GXMR-CAR Jurkat cells recognized soluble GXM from C. gattii and C. neoformans, and the levels of IL-2 released by the modified cells did not differ between the GXMR-CAR constructs after exposure to Cryptococcus spp. 18B7-GXMR-CAR triggered tonic signaling was more pronounced in modified Jurkat cells, and a protein kinase inhibitor of the Src family (dasatinib) significantly reduced GXMR-CAR tonic signaling and inhibited cell activation against ligands. 18B7 scFv showed a structural modification of the variable heavy (VH) chain that clarified the difference in the strength of tonic signaling and the level of cell activation between 2H1-GXMR-CAR and 18B7-GXMR-CAR. GXMR-CAR constructs induced T-cell activation against clinical isolates of Cryptococcus spp. and serum from patients with cryptococcosis induced high levels of IL-2, mainly in cells modified with 18B7-GXMR-CAR. Thus, 18B7-GXMR-CAR and 2H1-GXMR-CAR mediated T cell activation against Cryptococcus spp. and 18B7 and 2H1 scFv influenced the strength of tonic signaling. HIGHLIGHTS center dot 2H1-GXMR-CAR and 18B7-GXMR-CAR are efficiently expressed on the cell surface; center dot 2H1-GXMR-CAR and 18B7-GXMR-CAR redirected T cells toward the ligands; center dot 18B7-GXMR-CAR provided highest levels of tonic signaling; center dot Binding pocket of 18B7 scFv favored the tonic signaling triggered by GXMR-CAR; center dot Binding pocket of 18B7 scFv favored the tonic signaling triggered by GXMR-CAR; [GRAPHICS] . (AU) | |
| FAPESP's process: | 18/18538-0 - Bioengineered of T- and NK-cells by CAR against invasive fungal infections |
| Grantee: | Thiago Aparecido da Silva |
| Support Opportunities: | Research Grants - Young Investigators Grants |
| FAPESP's process: | 21/02758-4 - Construction of CAR receptors to redirect T and NK cells to control invasive Candida albicans infection |
| Grantee: | Gabriela Yamazaki de Campos |
| Support Opportunities: | Scholarships in Brazil - Doctorate |
| FAPESP's process: | 20/16738-2 - Redirection of T cells via CAR to fight against Candida albicans infection |
| Grantee: | Júlia Garcia Guimarães |
| Support Opportunities: | Scholarships in Brazil - Master |
| FAPESP's process: | 19/26074-7 - Bioengineered of T- and NK-cells by CAR against invasive fungal infections |
| Grantee: | Thiago Aparecido da Silva |
| Support Opportunities: | Scholarships in Brazil - Young Researchers |
| FAPESP's process: | 20/11307-3 - Impact of different scFv in GXMR-CAR expressed by T lymphocytes during T-cell activation against Cryptococcus spp |
| Grantee: | Michele Procópio Machado |
| Support Opportunities: | Scholarships in Brazil - Master |