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XU Weili, YYANG Xiaofeng, LI Meng, ZHOU Hui, LI Suolin. yeloid-derived Suppressor Cells Inhibit Proliferation and Killing Activity of Neuroblastoma Antigen-specific Cytotoxic T Lymphocyte in vitro[J]. Cancer Research on Prevention and Treatment, 2017, 44(11): 728-732. DOI: 10.3971/j.issn.1000-8578.2017.17.0709
Citation: XU Weili, YYANG Xiaofeng, LI Meng, ZHOU Hui, LI Suolin. yeloid-derived Suppressor Cells Inhibit Proliferation and Killing Activity of Neuroblastoma Antigen-specific Cytotoxic T Lymphocyte in vitro[J]. Cancer Research on Prevention and Treatment, 2017, 44(11): 728-732. DOI: 10.3971/j.issn.1000-8578.2017.17.0709

yeloid-derived Suppressor Cells Inhibit Proliferation and Killing Activity of Neuroblastoma Antigen-specific Cytotoxic T Lymphocyte in vitro

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  • Received Date: June 20, 2017
  • Revised Date: September 25, 2017
  • Available Online: January 12, 2024
  • Objective 

    To explore the inhibitory role of myeloid-derived suppressor cell (MDSC) in the proliferation and killing activity of neuroblastoma antigen-specific cytotoxic T lymphocyte (CTL) in vitro.

    Methods 

    The neuroblastoma antigen specific CTLs were successfully prepared on the basis of cultivation of neuroblastoma SK-N-SH cells and separation of BALB/c mice myeloid-derived dendritic cell (DC) and CD3+T cells in vitro. MDSCs were purified and cultivated with CTLs, then the inhibitory role of MDSC in the proliferation of CTL was detected by fluorescence staining of 5, 6-carboxyfluorescein diacetate succinimidyl ester (CFSE) and flow cytometry. Furthermore, CTL, SK-N-SH and MDSC were mixed and cultivated, the killing rate of CTL on SK-N-SH and the secretion of IL-2, IFN-γ in supernatant of the different groups were detected by ELISA.

    Results 

    After magnetic cell sorting, the rate of Gr-1+CD11b+MDSC reached to 84.6% by flow cytometry test. The levels of IL-2 and IFN-γ in supernatant of antigen-loaded CTLs were significantly higher than those in supernatant of T cells (P < 0.05). The proliferation of CTLs cultivated with MDSC was significantly inhibited, with strong fluorescence in view: however, CTLs cultivated alone proliferated obviously, with weak fluorescence intensity. The killing rate of CTLs to SK-N-SH in MDSC+CTL+SK-N-SH group was significant lower than that in CTL+SK-N-SH group (t=6.506, P < 0.001). Significant difference existed in the secretion levels of IL-2 and IFN-γ in the supernatant between the two groups (all P < 0.01).

    Conclusion 

    MDSC inhibite the proliferation and activity of neuroblastoma antigen-specific CTLs in vitro result in immune tolerance and reduced the killing effect of CTL on neuroblastoma cells.

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