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优化肿瘤相关抗原刺激的自体树突状细胞和细胞因子诱导的杀伤细胞共培养,以增强癌症免疫治疗中的细胞毒性。

Optimizing tumor-associated antigen-stimulated autologous dendritic cell and cytokine-induced killer cell coculture to enhance cytotoxicity for cancer immunotherapy in manufacturing.

机构信息

Division of Pediatric Neurosurgery, Department of Neurosurgery, Neurological Institute, Taipei Veterans General Hospital, Taipei, Taiwan.

School of Medicine, National Yang Ming Chiao Tung University, Taipei, Taiwan.

出版信息

BMC Immunol. 2023 Jun 29;24(1):14. doi: 10.1186/s12865-023-00552-5.

DOI:10.1186/s12865-023-00552-5
PMID:37386444
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10311787/
Abstract

BACKGROUND

Dendritic Cell Cytokine-induced killer cell (DC-CIK) coculture treatment in cancer immunotherapy has been shown to be effective. However, the cost of DC- CIK therapy is prohibitive for many patients, and the lack of standard manufacturing processes and treatment strategies are major limitations. Our study used tumor lysate as a tumor-associated antigen source and DCs and CIK cells in coculture. We developed an efficient method to obtain autologous DCs- and CIK cells from peripheral blood. We used flow cytometry to assess DC activation and the cytometric bead array assay to quantify cytokines secreted by CIK cells.

RESULTS

We evaluated the antitumor activity of DC- CIK coculture in vitro with the K562 cell line. We demonstrated that a manufacturing process employing frozen immature DCs can yield the lowest loss with the highest economic benefits. DC-CIK coculture can effectively upgrade CIK cells' immunological specificity to tumors in the presence of tumor-associated antigens.

CONCLUSION

In vitro experiments revealed that when the DC- CIK cell ratio was 1: 20 in the coculture, CIK cells secreted the highest number of cytokines on the 14th day and the antitumor immune effect showed the highest potency. CIK cells' cytotoxicity to K562 cells was highest when the CIK: K562 cell ratio was 25: 1. We developed an efficient manufacturing process for DC- CIK coculture, while also establishing the optimal DC- CIK cell ratio for immunological activity and the best cytotoxic CIK: K562 cell ratio.

摘要

背景

树突状细胞细胞因子诱导的杀伤细胞(DC-CIK)共培养治疗在癌症免疫治疗中已被证明是有效的。然而,对于许多患者来说,DC-CIK 治疗的费用是令人望而却步的,缺乏标准的制造工艺和治疗策略是主要的限制。我们的研究使用肿瘤裂解物作为肿瘤相关抗原的来源,并在共培养中使用 DC 和 CIK 细胞。我们开发了一种从外周血中获得自体 DC 和 CIK 细胞的有效方法。我们使用流式细胞术评估 DC 的激活,并用细胞因子珠阵列分析法来定量 CIK 细胞分泌的细胞因子。

结果

我们用 K562 细胞系评估了 DC-CIK 共培养的体外抗肿瘤活性。我们证明,采用冷冻未成熟 DC 的制造工艺可以在获得最高经济效益的同时,使损失最小化。在存在肿瘤相关抗原的情况下,DC-CIK 共培养可以有效地提高 CIK 细胞对肿瘤的免疫特异性。

结论

体外实验表明,当共培养中 DC-CIK 细胞比例为 1:20 时,CIK 细胞在第 14 天分泌的细胞因子数量最多,抗肿瘤免疫效应的效力最高。当 CIK:K562 细胞比例为 25:1 时,CIK 细胞对 K562 细胞的细胞毒性最高。我们开发了一种高效的 DC-CIK 共培养制造工艺,同时确定了免疫活性的最佳 DC-CIK 细胞比例和最佳的细胞毒性 CIK:K562 细胞比例。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a20b/10311787/0c90ec356bde/12865_2023_552_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a20b/10311787/e1e8350f4202/12865_2023_552_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a20b/10311787/9dc5228f0358/12865_2023_552_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a20b/10311787/d8876c906fd9/12865_2023_552_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a20b/10311787/0c90ec356bde/12865_2023_552_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a20b/10311787/e1e8350f4202/12865_2023_552_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a20b/10311787/9dc5228f0358/12865_2023_552_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a20b/10311787/d8876c906fd9/12865_2023_552_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a20b/10311787/0c90ec356bde/12865_2023_552_Fig4_HTML.jpg

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