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The effects of αGalCer-induced TCRVα24 Vβ11+ natural killer T cells on NK cell cytotoxicity in umbilical cord blood

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Abstract

Purpose

The first objective of this study was to investigate in vitro effects of α-galactosylceramide (αGalCer) on the proliferation of umbilical cord blood (UCB) natural killer T (NKT) cells and enhancement of their cytotoxicity. The second one is to examine whether purified NKT cells could affect the cytotoxicity of UCB-NK cells either in the presence or absence of dendritic cells (DCs).

Methods

Mononuclear cells (MNCs) from UCB were cultured for 2 weeks in the presence of IL-2 (100 U/ml), with or without αGalCer. The effect of neutralizing monoclonal antibodies (MoAb) against TCRVα24 and CD1d was also examined. TCRVα24 Vβ11 double positive NKT cells were purified by FACS sorter and then cocultured with syngeneic isolated UCB−CD56+NK cells in either the presence or absence of DCs. The cytotoxicity against various malignant cell targets and cytokine production was determined.

Results

The addition of αGalCer induced human NKT cells to proliferate in UCB-MNCs to a greater extent than in adult PB-MNCs. However, it suppressed the cytotoxic activity against malignant cell targets. Anti-TCRVα24 and CD1d MoAb recovered the cytotoxicity by inhibiting the proliferation of UCB-NKT cells. NKT cells cocultured with auto-DCs significantly increased NK cell cytotoxicity against K562, and Raji cells and produced IFN-γ at much higher levels than UCB-NKT cells alone.

Conclusion

In UCB samples, αGalCer–pulsed DCs and NKT cells acted together to enhance NK cytotoxicity in vitro.

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Acknowledgements

This work was supported by Health and Labor Sciences Research Grants, Research on Pharmaceutical and Medical Safety, and the Japanese Health Sciences Foundation, Research on Health Sciences focusing on Drug Innovation. We particularly wish to thank Dr William J Hubbard (University of Alabama at Birmingham, AL, USA) for comments.

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Authors and Affiliations

  1. Department of Hematology and Oncology, Tokai University School of Medicine, Bohseidai, 259-1193, Isehara, Kanagawa, Japan

    Yoko Ueda, Masao Hagihara, Ayako Okamoto, Kei Tazume & Tomomitsu Hotta

  2. Department of Cell Transplantation and Regenerative Medicine, Tokai University, 259-1193, Isehara, Kanagawa, Japan

    Aya Masui, Ayako Higuchi & Syunichi Kato

  3. Division of Transplant of Immunology, University of Alabama at Birmingham, Birmingham, Alabama, USA

    Balgansuren Gansuvd

  4. Division of Pulmonary and Critical Care Medicine, Department of Medicine, University of California at San Diego Medical Center, San Diego, California, USA

    Ying Yu

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  1. Yoko Ueda
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Correspondence to Masao Hagihara.

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Ueda, Y., Hagihara, M., Gansuvd, B. et al. The effects of αGalCer-induced TCRVα24 Vβ11+ natural killer T cells on NK cell cytotoxicity in umbilical cord blood. Cancer Immunol Immunother 52, 625–631 (2003). https://doi.org/10.1007/s00262-003-0398-3

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