PDF(Pubmed)
Abstract：
BACKGROUND: 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.
METHODS: 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.
RESULTS: 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.
CONCLUSIONS: 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.
METHODS: 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.
RESULTS: 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.
CONCLUSIONS: 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.
摘要：
背景：过继性癌症免疫治疗，使用工程化的T细胞，表达嵌合抗原受体或自体肿瘤浸润淋巴细胞，近年来,不同类型癌症的主要治疗方法。然而,尽管过继性癌症免疫疗法具有转化潜力，这个领域仍然面临重大挑战，表现为效应CD8+T细胞的细胞毒性能力在其扩增后明显下降。为了应对这些挑战，我们开发了一种离体“合成免疫生态位”(SIN)，由固定的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，甚至增强它们的细胞毒性。
方法：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细胞的增殖和细胞毒性之间的相互作用的分子机制，我们通过流式细胞术对特异性分化标志物进行了综合分析，以及基因表达谱分析。
结果:在第3天,对于树突细胞(DC)/卵清蛋白(OVA)活化的和抗CD3/CD28活化的细胞,由SIN诱导的转录组效应在很大程度上相似。细胞增殖增加，细胞表现出高杀伤能力。在第四天和以后，增殖/细胞毒性表型完全变成“激活特异性”；DC/OVA激活的细胞失去其细胞毒活性，which,反过来,被SIN治疗救出。在更长的孵化时间里，细胞毒活性进一步下降，在第7天，不能被罪拯救。用抗CD3/CD28珠活化后的SIN刺激诱导增殖表型的主要增加,同时瞬时抑制其细胞毒性2-3天,并在第7天完全恢复其杀伤活性。确定了SIN效应的潜在分子调控途径，基于转录组学和多光谱成像分析。
结论：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到初始激活模式的不同调节。SIN刺激大大增强了细胞的扩增，在两种激活模式下，在刺激后的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加强过继性癌症免疫疗法。
方法：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细胞的增殖和细胞毒性之间的相互作用的分子机制，我们通过流式细胞术对特异性分化标志物进行了综合分析，以及基因表达谱分析。
结果:在第3天,对于树突细胞(DC)/卵清蛋白(OVA)活化的和抗CD3/CD28活化的细胞,由SIN诱导的转录组效应在很大程度上相似。细胞增殖增加，细胞表现出高杀伤能力。在第四天和以后，增殖/细胞毒性表型完全变成“激活特异性”；DC/OVA激活的细胞失去其细胞毒活性，which,反过来,被SIN治疗救出。在更长的孵化时间里，细胞毒活性进一步下降，在第7天，不能被罪拯救。用抗CD3/CD28珠活化后的SIN刺激诱导增殖表型的主要增加,同时瞬时抑制其细胞毒性2-3天,并在第7天完全恢复其杀伤活性。确定了SIN效应的潜在分子调控途径，基于转录组学和多光谱成像分析。
结论：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到初始激活模式的不同调节。SIN刺激大大增强了细胞的扩增，在两种激活模式下，在刺激后的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加强过继性癌症免疫疗法。
