Optimization of Culture Conditions for the Generation of Canine CD20-CAR-T Cells for Adoptive Immunotherapy.

Osamu Sakai
Hiroka Yamamoto
Masaya Igase
Takuya Mizuno

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.

In vivo (Athens, Greece)

36

2

764

772

10.21873/invivo.12763

https://doi.org/10.21873/invivo.12763
https://www.ncbi.nlm.nih.gov/pubmed/35241532
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8931891