[摘 要] 目的：探讨分化抑制因子2（Id2）在诱导生成中央记忆性T（Tcm）细胞及增强T细胞抗肿瘤持久性中的作用。方法： 磁珠分选CD8+初始T细胞，与负载癌胚抗原（CEA）的树突状细胞（DC）共培养，经白介素-2（IL-2）或IL-7/15/21/23分别诱导培养 效应T（Teff）或Tcm细胞；qPCR和WB法分别检测T细胞中Id2和Id3 mRNA、蛋白表达；慢病毒敲减T细胞中Id2基因，用流式细 胞术检测其T细胞记忆表型；WB法检测PI3K/AKT通路相关蛋白的表达；Seahorse能量代谢仪分析细胞外酸化速率（ECAR）和耗 氧速率（OCR）；斑马鱼结肠癌HCT116细胞移植瘤模型分析Teff和Tcm细胞的抗肿瘤差异，进一步观察敲减Id2基因的Tcm细胞 （Tcm-shId2）对第二次移植瘤的生长抑制。结果：Tcm细胞高表达Id3 mRNA（P < 0.05），而Teff细胞高表达Id2 mRNA（P < 0.001）。 成功构建敲减Id2基因的Tcm细胞（Tcm-shId2）且其Id3表达明显上调，敲减Id2可促进Tcm细胞的形成（P < 0.05）。Tcm-shId2细 胞通过PI3K/AKT通路进行代谢重编程，有效抑制斑马鱼体内结肠癌移植瘤的生长，对第二次移植瘤也能产生显著抑制作用 （P < 0.01）。结论：Id2可能通过调控PI3K/AKT通路改变T细胞代谢模式，从而促进CD8+ T细胞向Tcm细胞分化，有效抑制结肠 癌移植瘤的生长。

[Abstract] Objective: To investigate the role of inhibitor of differentiation 2 (Id2) in inducing the generation of central memory T (Tcm) cells and enhancing the anti-tumor persistence of T cells. Methods: CD8+ na?ve T cells were sorted with magnetic beads and then co-cultured with carcinoembryonic antigen (CEA)-loaded dendritic cells (DCs). These cells were induced into effector T (Teff) or Tcm cells by interleukin-2 (IL-2) or IL-7/15/21/23, respectively. The mRNA and protein expression of Id2 and Id3 in T cells were detected using qPCR and WB, respectively. Id2 gene in T cells was knocked down using lentivirus, and the T cell memory phenotype was analyzed by flow cytometry. The expression of PI3K/AKT pathway-related proteins was examined by WB. The extracellular acidification rate (ECAR) and oxygen consumption rate (OCR) were assessed using a Seahorse extracellular flux analyzer. A zebrafish colorectal cancer HCT116 xenograft model was employed to analyze the anti-tumor differences between Teff and Tcm cells. The effect of Id2 gene knockdown in Tcm cells (Tcm-shId2) on the growth inhibition of secondary xenografts was also observed. Results: Tcm cells exhibited high expression of Id3 mRNA (P < 0.05), whereas Teff cells showed high expression of Id2 mRNA (P < 0.001). Tcm cells with Id2 knockdown (Tcm-shId2) were successfully constructed, showing significantly upregulated Id3 expression. Knockdown of Id2 promoted the formation of Tcm cell (P < 0.05). Tcm-shId2 cells underwent metabolic reprogramming via the PI3K/AKT pathway, which effectively suppressed the growth of colorectal cancer xenografts in zebrafish and also produced significant inhibitory effects on secondary tumor growth (P < 0.01). Conclusion: Id2 gene may regulate T cell metabolism through the PI3K/AKT signaling pathway,promoting the differentiation of CD8+ T cells into Tcm cells and effectively inhibiting the growth of colorectal cancer xenografts.

福建省卫生计生科研人才培养项目医学创新课题（No. 2018-CX-9）；福建省科技创新联合资金（No. 2018Y9108）； 福建省自然科学基金（No. 2023J011247）；福建省科技创新联合资金（No. 2024Y9607）