优化犬源 CD20-CAR-T 细胞培养条件用于过继免疫治疗。
Optimization of Culture Conditions for the Generation of Canine CD20-CAR-T Cells for Adoptive Immunotherapy.
机构信息
Laboratory of Molecular Diagnostics and Therapeutics, Joint Graduate school of Veterinary Medicine, Yamaguchi University, Yoshida, Japan.
Laboratory of Molecular Diagnostics and Therapeutics, Joint Graduate school of Veterinary Medicine, Yamaguchi University, Yoshida, Japan
出版信息
In Vivo. 2022 Mar-Apr;36(2):764-772. doi: 10.21873/invivo.12763.
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.
背景/目的：靶向 CD20 的嵌合抗原受体 (CAR) T 细胞疗法有可能成为治疗犬 B 细胞淋巴样恶性肿瘤的一种有前途的新方法。然而，对于产生具有犬类有利表型的有效 CAR-T 细胞的最佳方法仍不清楚。在这项研究中，我们评估了几种培养条件及其对 CD20-CAR-T 细胞表型特征的影响。
材料和方法
通过在存在或不存在 Akt 抑制剂的情况下用几种有丝分裂原孵育来产生犬科 CAR-T 细胞。通过流式细胞术确定基因转导效率和表型特征。
结果
比较几种有丝分裂原发现，植物血球凝集素刺激具有高转导效率，而刀豆蛋白 A 刺激在记忆 T 细胞形成方面更具优势。在 CAR-T 生产的初始阶段抑制 Akt 倾向于提高转导效率和记忆 T 细胞的形成。
结论
本研究为理解在过继免疫治疗中犬 T 细胞的体外扩增提供了重要的见解。
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