| 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. |
| 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 |
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| KeyWord:glioblastoma; immunotherapy; dendritic cell (DC) vaccine; reactive oxygen species |
| 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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| Abstract: |
| 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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