| Xiaojin Liu,Yiwei Qi (Co-first author),Feng Hu,Kai Shu,Ting Lei. Differential centrifugation enhances the anti-tumor immune effect of tumor lysate-pulsed dendritic cell vaccine against glioblastoma. Oncol Transl Med, 2022, 8: 209-216. |
| 差速离心增强全肿瘤抗原致敏树突状细胞疫苗体外抗肿瘤免疫活性的研究 |
| Differential centrifugation enhances the anti-tumor immune effect of tumor lysate-pulsed dendritic cell vaccine against glioblastoma |
| Received:June 15, 2022 Revised:October 21, 2022 |
| DOI：10.1007/s10330-022-0582-2 |
| 中文关键词: 胶质母细胞瘤；免疫治疗；树突状细胞疫苗；活性氧 |
| 英文关键词: glioblastoma; immunotherapy; dendritic cell (DC) vaccine; reactive oxygen species |
| 基金项目: |
| Author Name | Affiliation | E-mail | | Xiaojin Liu | Sino-German Neuro-Oncology Molecular Laboratory, Department of Neurosurgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology | liuxiaojin@tjh.tjmu.edu.cn | | Yiwei Qi (Co-first author) | Sino-German Neuro-Oncology Molecular Laboratory, Department of Neurosurgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology | | | Feng Hu | Sino-German Neuro-Oncology Molecular Laboratory, Department of Neurosurgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology | | | Kai Shu | Sino-German Neuro-Oncology Molecular Laboratory, Department of Neurosurgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology | | | Ting Lei* | Sino-German Neuro-Oncology Molecular Laboratory, Department of Neurosurgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology | tlei@tjh.tjmu.edu.cn |
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| 中文摘要: |
| 目的：本研究旨在通过差速离心提高全肿瘤抗原致敏树突状细胞疫苗的体外抗肿瘤免疫活性，为其临床应用于胶质母细胞瘤提供理论依据。方法：我们使用Ficoll-Paque PLUS提取外周血单个核细胞，并在体外用细胞因子将它们诱导成成熟的树突状细胞。通过差速离心制备改良全肿瘤抗原。流式细胞仪检测各组树突状细胞成熟标志物，包括改良全肿瘤抗原、全肿瘤抗原、阴性和阳性对照组。此外，它们刺激淋巴细胞增殖的能力和体外抗肿瘤作用通过 Cell Trace TM CFSE 进行了评估。通过ELISA测量IFN-γ分泌水平。通过 2",7"-二氯荧光素二乙酸酯 (DCFDA) 染色测量细胞内活性氧。所有结果均采用unpaired Student"s t-test进行统计学比较，P < 0.05 具有统计学差异。结果：与全肿瘤抗原致敏的DCs相比，改良全肿瘤抗原致敏DCs具有更高的成熟标志物表达：CD1a（7.38±0.53% VS 4.47±0.75%）和CD83（19.81±4.09% VS 9.64±1.50%），更能刺激淋巴细胞增殖（增殖指数/PI：8.54±0.16 VS 7.35±0.05）和分泌IFN-γ并诱导更强的体外对胶质母细胞瘤细胞的CTLs细胞毒性。此外，我们发现改良全肿瘤抗原致敏 DCs 内的 ROS 水平低于全肿瘤抗原致敏 DCs。结论：差速离心法可提高全肿瘤抗原致敏树突状细胞疫苗的体外抗肿瘤免疫活性，活性氧可能是影响全肿瘤抗原致敏树突状细胞功能的关键。 |
| 英文摘要: |
| Objective This study aimed to improve the antitumor immunocompetence of a tumor lysate-pulsed
dendritic cell (DC) vaccine through differential centrifugation and provide a theoretical basis for its clinical
application in glioblastoma.
Methods Peripheral blood mononuclear cells were extracted using Ficoll-Paque PLUS and induced
into mature DCs in vitro with a cytokine cocktail. The modified tumor lysate was generated by differential
centrifugation. The maturity mar kers of DCs in each group, namely the modified tumor lysate, tumor lysate,
and negative and positive control groups, were assessed using flow cytometry. Furthermore, their ability to
stimulate lymphocyte proliferation and in vitro antitumor effects were assessed using Cell Trace TM CFSE.
IFN-γ secretion levels were measured with ELISA. Intracellular reactive oxygen species were measured
using 2’,7’-dichlorofluorescein diacetate (DCFDA) staining. The results were statistically analyzed using an
unpaired Student’s t-test and were considered significant at P < 0.05.
Results Compared with tumor lysate-pulsed DCs, modified tumor lysate-pulsed DCs had a higher
expression of maturity markers: CD1a (7.38 ± 0.53% vs. 4.47 ± 0.75%) and CD83 (19.81 ± 4.09% vs. 9.64
± 1.50%), were better capable of stimulating lymphocyte proliferation [proliferation index (PI): 8.54 ± 0.16
vs. 7.35 ± 0.05], secreting IFN-γ, and inducing stronger in-vitro cytotoxic T lymphocyte (CTL) cytotoxicity
against glioblastoma cells. In addition, we found that the level of ROS in modified tumor lysate-pulsed DCs
was lower than that in tumor lysate-pulsed DCs.
Conclusion Differential centrifugation of tumor lysates can improve the antitumor immunocompetence
of DC vaccines, and reactive oxygen species may be the key to affecting DC function in the whole tumor
lysate. |
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