Analysis and Development of Antigen-specific T Cells Derived from Peripheral Blood Mononuclear Cells of Healthy Donors.
Anticancer Res
; 44(4): 1377-1387, 2024 Apr.
Article
in En
| MEDLINE
| ID: mdl-38537976
ABSTRACT
BACKGROUND/AIM:
Adoptive cell therapy using antigen-specific T cells is a promising treatment modality for cancer patients. Various methods to isolate specific T cells and identify corresponding T cell receptor (TCR) sequences are known. This study aimed to identify antigen-specific TCR from T cells isolated using carboxyfluorescein succinimidyl ester (CFSE), which marks proliferating activated T cells. MATERIALS ANDMETHODS:
CFSE stained healthy donor peripheral blood mononuclear cells (PBMCs) were treated with cytomegalovirus (CMV) or Epstein-Barr virus (EBV) peptides for seven days. Then, proliferating T cells with decreased CFSE staining were isolated and single cell VDJ sequencing was performed on isolated T cells to identify antigen-specific TCRs.RESULTS:
As antigen-specific TCR candidates, ten TCR clones were selected for the CMV antigen and five for the EBV antigen. The reactivity of ten CMV TCR-transduced T cells and one EBV TCR-transduced T cell toward T2 cells pulsed with CMV or EBV peptide was confirmed via NFAT-luciferase, IFN-γ ELISA, and cytotoxicity assays.CONCLUSION:
Identification of antigen-specific TCRs with CFSE staining is a valid method for the development of effective immunotherapy. The identified CMV- or EBV-specific TCRs can be used for adoptive cell therapy to treat cancer.Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Succinimides
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Cytomegalovirus Infections
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Epstein-Barr Virus Infections
/
Fluoresceins
/
Neoplasms
Limits:
Humans
Language:
En
Journal:
Anticancer Res
Year:
2024
Document type:
Article