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外周血自然杀伤(NK)细胞在人工抗原呈递细胞存在或不存在的情况下培养时的扩增比较的实验研究。

An Experimental Study Comparing the Expansion of Peripheral Blood Natural Killer (NK) Cells Cultured with Artificial Antigen-Presenting Cells, in the Presence or Absence of Bone Marrow Mesenchymal Stem Cells (MSCs).

机构信息

Cellular Technology and Therapy Center, Hospital de Clínicas de Porto Alegre, Rua Ramiro Barcelos, 2350, Porto Alegre, RS, CEP 90035-903, Brazil.

Federal University of Rio Grande Do Sul, Porto Alegre, RS, Brazil.

出版信息

Mol Biotechnol. 2020 May;62(5):306-315. doi: 10.1007/s12033-020-00250-2.

Abstract

NK cells have been seen as potential agents in adoptive immunotherapy for cancer. The main challenge for the success of this approach is to obtain a great quantity of activated NK cells for adoptive transfer. The present study had aimed to evaluate the effect of a feeder layer of irradiated MSCs in the in vitro expansion of NK cells. MSCs were obtained from the bone marrow (BM) cells remaining in the bag and filter used in the transplantation of hematopoietic stem cells. NK cells were obtained from peripheral blood (PB) of healthy volunteers. NK expansion and activation were stimulated by culture with artificial antigen-presenting cells (aAPCs) and IL-2, in the presence or absence of BM-MSCs. NK cell proliferation, phenotypic expression and cytotoxic activity were evaluated. Both culture conditions showed high NK purity with predominance of NK CD56CD16 subset post expansion. However, cultures without the presence of MSCs showed higher NK proliferation, expression of activation markers (CD16 and NKG2D) and related cytotoxic activity. In this experimental study, the presence of a feeder layer of irradiated BM-MSCs interfered negatively in the expansion of PB-NKs, limiting their growth and activation. Further investigation is needed to understand the mechanisms of NK-MSC interaction and its implications.

摘要

自然杀伤 (NK) 细胞被视为癌症过继免疫治疗的潜在药物。这种方法成功的主要挑战是获得大量激活的 NK 细胞进行过继转移。本研究旨在评估辐照骨髓间充质干细胞 (MSCs) 饲养层对 NK 细胞体外扩增的影响。MSCs 从造血干细胞移植中剩余的袋子和过滤器中的骨髓 (BM) 细胞中获得。NK 细胞从健康志愿者的外周血 (PB) 中获得。通过与人工抗原呈递细胞 (aAPC) 和 IL-2 培养,在存在或不存在 BM-MSCs 的情况下刺激 NK 细胞扩增和激活。评估了 NK 细胞的增殖、表型表达和细胞毒性活性。两种培养条件均显示 NK 纯度高,扩增后 NK CD56CD16 亚群占主导地位。然而,没有 MSCs 存在的培养显示出更高的 NK 增殖、激活标志物 (CD16 和 NKG2D) 的表达和相关的细胞毒性活性。在这项实验研究中,辐照 BM-MSCs 饲养层的存在对 PB-NK 的扩增产生了负面影响,限制了它们的生长和激活。需要进一步研究以了解 NK-MSC 相互作用的机制及其影响。

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