Department of Immunology and Regenerative Biology, Weizmann Institute of Science, Rehovot, Israel.
Bioinformatics Unit, Department of Life Sciences Core Facilities, Weizmann Institute of Science, Rehovot, Israel.
J Immunother Cancer. 2024 Jun 12;12(6):e009011. doi: 10.1136/jitc-2024-009011.
Adoptive cancer immunotherapy, using engineered T-cells, expressing chimeric antigen receptor or autologous tumor infiltrating lymphocytes became, in recent years, a major therapeutic approach for diverse types of cancer. However, despite the transformative potential of adoptive cancer immunotherapy, this field still faces major challenges, manifested by the apparent decline of the cytotoxic capacity of effector CD8 T cells upon their expansion. To address these challenges, we have developed an ex vivo "synthetic immune niche" (SIN), composed of immobilized CCL21 and ICAM1, which synergistically induce an efficient expansion of antigen-specific CD8 T cells while retaining, and even enhancing their cytotoxic potency.
To explore the molecular mechanisms through which a CCL21+ICAM1-based SIN modulates the interplay between the proliferation and cytotoxic potency of antigen-activated and CD3/CD28-activated effector CD8 T cells, we performed integrated analysis of specific differentiation markers via flow cytometry, together with gene expression profiling.
On day 3, the transcriptomic effect induced by the SIN was largely similar for both dendritic cell (DC)/ovalbumin (OVA)-activated and anti-CD3/CD28-activated cells. Cell proliferation increased and the cells exhibited high killing capacity. On day 4 and on, the proliferation/cytotoxicity phenotypes became radically "activation-specific"; The DC/OVA-activated cells lost their cytotoxic activity, which, in turn, was rescued by the SIN treatment. On longer incubation, the cytotoxic activity further declined, and on day7, could not be rescued by the SIN. SIN stimulation following activation with anti-CD3/CD28 beads induced a major increase in the proliferative phenotype while transiently suppressing their cytotoxicity for 2-3 days and fully regaining their killing activity on day 7. Potential molecular regulatory pathways of the SIN effects were identified, based on transcriptomic and multispectral imaging profiling.
These data indicate that cell proliferation and cytotoxicity are negatively correlated, and the interplay between them is differentially regulated by the mode of initial activation. The SIN stimulation greatly enhances the cell expansion, following both activation modes, while displaying high survival and cytotoxic potency at specific time points following stimulation, suggesting that it could effectively reinforce adoptive cancer immunotherapy.
近年来，采用嵌合抗原受体或自体肿瘤浸润淋巴细胞的过继性癌症免疫疗法已成为多种癌症的主要治疗方法。然而，尽管过继性癌症免疫疗法具有变革性的潜力，但该领域仍面临重大挑战，表现为效应 CD8 T 细胞在扩增后细胞毒性明显下降。为了解决这些挑战，我们开发了一种体外“合成免疫生态位”（SIN），由固定化 CCL21 和 ICAM1 组成，它们协同诱导抗原特异性 CD8 T 细胞的有效扩增，同时保留甚至增强其细胞毒性。
为了探索基于 CCL21+ICAM1 的 SIN 调节抗原激活和 CD3/CD28 激活的效应 CD8 T 细胞增殖和细胞毒性之间相互作用的分子机制，我们通过流式细胞术进行了特定分化标志物的综合分析，同时进行了基因表达谱分析。
在第 3 天，SIN 诱导的转录组效应对于树突状细胞（DC）/卵清蛋白（OVA）激活和抗 CD3/CD28 激活的细胞大致相似。细胞增殖增加，细胞具有高杀伤能力。在第 4 天及以后，增殖/细胞毒性表型变得截然不同“激活特异性”；DC/OVA 激活的细胞失去了细胞毒性，而 SIN 处理则挽救了这种细胞毒性。在更长的孵育时间后，细胞毒性进一步下降，在第 7 天，无法通过 SIN 挽救。用抗 CD3/CD28 珠激活后用 SIN 刺激会引起增殖表型的显著增加，同时短暂抑制其细胞毒性 2-3 天，并在第 7 天完全恢复其杀伤活性。基于转录组和多光谱成像谱分析，确定了 SIN 效应的潜在分子调控途径。
这些数据表明细胞增殖和细胞毒性呈负相关，它们之间的相互作用受初始激活模式的差异调节。SIN 刺激极大地增强了两种激活模式后的细胞扩增，同时在刺激后特定时间点显示出高存活率和细胞毒性，这表明它可以有效地增强过继性癌症免疫疗法。
