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- Title
Optimization of Culture Conditions for the Generation of Canine CD20-CAR-T Cells for Adoptive Immunotherapy.
- Authors
OSAMU SAKAI; HIROKA YAMAMOTO; MASAYA IGASE; TAKUYA MIZUNO
- Abstract
Background/Aim: Chimeric antigen receptor (CAR) T cell therapy targeting CD20 has the potential to become a promising novel treatment for canine B cell lymphoid malignancy. However, the optimal approach for producing potent CAR-T cells with favorable phenotype for dogs remains unknown. In this study, we assessed several culture conditions and their effects on the phenotypic characteristics of CD20-CAR-T cells. Materials and Methods: Canine CAR-T cells were generated by incubating with several mitogens in the presence or absence of Akt inhibitor. Gene transduction efficiency and phenotypic characteristics were determined by flow cytometry. Results: Comparison of several kinds of mitogens revealed that stimulation with phytohemagglutinin has high transduction efficacy, whereas stimulation with concanavalin A was superior in memory T cell formation. Akt inhibition at the initial stage of CAR-T production tended to enhance transduction efficiency and memory T cell formation. Conclusion: This study provides a significant insight into the understanding of the ex vivo expansion of canine T cells in adoptive immunotherapy.
- Subjects
CHIMERIC antigen receptors; B cell lymphoma; IMMUNOTHERAPY; PHYTOHEMAGGLUTININS; CELLULAR therapy; DOG diseases
- Publication
In Vivo, 2022, Vol 36, Issue 2, p764
- ISSN
0258-851X
- Publication type
Article
- DOI
10.21873/invivo.12763