PDF(Pubmed)
Abstract：
UNASSIGNED: Medulloblastoma (MB) is the most prevalent paediatric brain tumour. Despite improvements in patient survival with current treatment strategies, the quality of life of these patients remains poor owing to the sequelae and relapse risk. An alternative, or, in addition to the current standard treatment, could be considered immunotherapy, such as Natural Killer cells (NK). NK cells are cytotoxic innate lymphoid cells that play a major role in cancer immunosurveillance. To date, the mechanism of cytotoxicity of NK cells, especially regarding the steps of adhesion, conjugation, cytotoxic granule polarisation in the cell contact area, perforin and granzyme release in two and three dimensions, and therapeutic efficacy in vivo have not been precisely described.
UNASSIGNED: Each step of NK cytotoxicity against the three MB cell lines was explored using confocal microscopy for conjugation, Elispot for degranulation, flow cytometry, and luminescence assays for target cell necrosis and lysis and mediators released by cytokine array, and then confirmed in a 3D spheroid model. Medulloblastoma-xenografted mice were treated with NK cells. Their persistence was evaluated by flow cytometry, and their efficacy in tumour growth and survival was determined. In addition, their effects on the tumour transcriptome were evaluated.
UNASSIGNED: NK cells showed variable affinities for conjugation with MB target cells depending on their subgroup and cytokine activation. Chemokines secreted during NK and MB cell co-culture are mainly associated with angiogenesis and immune cell recruitment. NK cell cytotoxicity induces MB cell death in both 2D and 3D co-culture models. NK cells initiated an inflammatory response in a human MB murine model by modulating the MB cell transcriptome.
UNASSIGNED: Our study confirmed that NK cells possess both in vitro and in vivo cytotoxic activity against MB cells and are of interest for the development of immunotherapy.
UNASSIGNED: Each step of NK cytotoxicity against the three MB cell lines was explored using confocal microscopy for conjugation, Elispot for degranulation, flow cytometry, and luminescence assays for target cell necrosis and lysis and mediators released by cytokine array, and then confirmed in a 3D spheroid model. Medulloblastoma-xenografted mice were treated with NK cells. Their persistence was evaluated by flow cytometry, and their efficacy in tumour growth and survival was determined. In addition, their effects on the tumour transcriptome were evaluated.
UNASSIGNED: NK cells showed variable affinities for conjugation with MB target cells depending on their subgroup and cytokine activation. Chemokines secreted during NK and MB cell co-culture are mainly associated with angiogenesis and immune cell recruitment. NK cell cytotoxicity induces MB cell death in both 2D and 3D co-culture models. NK cells initiated an inflammatory response in a human MB murine model by modulating the MB cell transcriptome.
UNASSIGNED: Our study confirmed that NK cells possess both in vitro and in vivo cytotoxic activity against MB cells and are of interest for the development of immunotherapy.
摘要：
■髓母细胞瘤（MB）是最常见的儿科脑肿瘤。尽管目前的治疗策略改善了患者的生存率，由于后遗症和复发风险，这些患者的生活质量仍然很差。另一种选择，或者，除了目前的标准治疗，可以考虑免疫疗法，例如自然杀伤细胞(NK)。NK细胞是细胞毒性的固有淋巴细胞，在癌症免疫监视中起主要作用。迄今为止,NK细胞的细胞毒性机制，特别是关于粘合的步骤，共轭，细胞接触区域的细胞毒性颗粒极化，穿孔素和颗粒酶在二维和三维中的释放，和体内治疗功效尚未得到准确描述。
■使用共聚焦显微镜进行接合，探索了针对三种MB细胞系的NK细胞毒性的每个步骤，脱粒的elispot,流式细胞术,以及通过细胞因子阵列释放的靶细胞坏死和裂解以及介质的发光测定，然后在3D球体模型中确认。用NK细胞处理髓母细胞瘤异种移植小鼠。通过流式细胞术评估它们的持久性，并确定了它们在肿瘤生长和存活中的功效。此外,评估了它们对肿瘤转录组的影响。
■NK细胞显示与MB靶细胞结合的不同亲和力,这取决于它们的亚群和细胞因子活化。NK和MB细胞共培养过程中分泌的趋化因子主要与血管生成和免疫细胞募集有关。NK细胞的细胞毒性在2D和3D共培养模型中诱导MB细胞死亡。NK细胞通过调节MB细胞转录组在人MB鼠模型中引发炎症反应。
■我们的研究证实，NK细胞在体外和体内都具有针对MB细胞的细胞毒活性，并对免疫治疗的发展感兴趣。
■使用共聚焦显微镜进行接合，探索了针对三种MB细胞系的NK细胞毒性的每个步骤，脱粒的elispot,流式细胞术,以及通过细胞因子阵列释放的靶细胞坏死和裂解以及介质的发光测定，然后在3D球体模型中确认。用NK细胞处理髓母细胞瘤异种移植小鼠。通过流式细胞术评估它们的持久性，并确定了它们在肿瘤生长和存活中的功效。此外,评估了它们对肿瘤转录组的影响。
■NK细胞显示与MB靶细胞结合的不同亲和力,这取决于它们的亚群和细胞因子活化。NK和MB细胞共培养过程中分泌的趋化因子主要与血管生成和免疫细胞募集有关。NK细胞的细胞毒性在2D和3D共培养模型中诱导MB细胞死亡。NK细胞通过调节MB细胞转录组在人MB鼠模型中引发炎症反应。
■我们的研究证实，NK细胞在体外和体内都具有针对MB细胞的细胞毒活性，并对免疫治疗的发展感兴趣。
