The effects of αGalCer-induced TCRVα24 Vβ11+ natural killer T cells on NK cell cytotoxicity in umbilical cord blood
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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).
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.
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.
In UCB samples, αGalCer–pulsed DCs and NKT cells acted together to enhance NK cytotoxicity in vitro.
KeywordsUmbilical cord blood αGalCer Natural killer T cells NK cells Dendritic cells
- 5.Brooks EG., Balk SP, Aupeix K, Colonna M, Strominger JL, Groh-Spies V (1993) Human T-cell receptor (TCR) alpha/beta + CD4-CD8- T cells express oligoclonal TCRs, share junctional motifs across TCR V beta-gene families, and phenotypically resemble memory T cells. Proc Natl Acad Sci USA 90:11787PubMedGoogle Scholar
- 10.Nicol A, Nieda M, Koezuka Y, Porcelli S, Suzuki K, Tadokoro K, Durrant S, Juji T (2000) Human invariant Valpha24+ natural killer T cells activated by alpha-galactosylceramide (KRN7000) have cytotoxic anti-tumour activity through mechanisms distinct from T cells and natural killer cells. Immunology 99:229CrossRefPubMedGoogle Scholar
- 12.Hagihara M, Gansuvd B, Ueda Y, Tsuchiya T, Masui A, Tazume K, Inoue H., Kato S, Hotta T. (2002) Killing activity of human umbilical cord blood derived TCRValpha 24+ NKT cells against normal and malignant hematological cells in vitro: a comparative study with NK cells or OKT3 activated T lymphocytes or with adult peripheral blood NKT cells. Cancer Immunol Immunother 51:1CrossRefPubMedGoogle Scholar
- 13.Kitamura H, Iwakabe K, Yahata T, Nishimura S, Ohta A, Ohmi Y, Sato M, Takeda K, Okumura K, Van Kaer L, Kawano T, Taniguchi M, Nishimura T (1999) The natural killer T (NKT) cell ligand alpha-galactosylceramide demonstrates its immunopotentiating effect by inducing interleukin (IL)-12 production by dendritic cells and IL-12 receptor expression on NKT cells. J Exp Med 189:1121PubMedGoogle Scholar
- 16.Ishihara S, Nieda M, Kitayama J, Osada T, Yabe T, Kikuchi A, Koezuka Y, Porcelli SA, Tadokoro K, Nagawa H, Juji T (2000) Alpha-glycosylceramides enhance the antitumor cytotoxicity of hepatic lymphocytes obtained from cancer patients by activating CD3−CD56+ NK cells in vitro. J Immunol 165:1659PubMedGoogle Scholar
- 23.Poggi A, Carosio R, Spaggiari GM, Fortis C, Tambussi G, Dell'Antonio G, Dal Cin E, Rubartelli A, Zocchi MR (2002) NK cell activation by dendritic cells is dependent on LFA-1-mediated induction of calcium-calmodulin kinase II: inhibition by HIV-1 Tat C-terminal domain. J Immunol 168:95PubMedGoogle Scholar
- 30.Okai M, Nieda M, Tazbirkova A, Horley D, Kikuchi A, Durrant S, Takahashi T,. Boyd A, Abraham R, Yagita H, Juji T, Nicol A (2002) Human peripheral blood Valpha24+ Vbeta11+ NKT cells expand following administration of alpha-galactosylceramide-pulsed dendritic cells. Vox Sang 83:250CrossRefPubMedGoogle Scholar
- 41.Hammond K, Poulton LD, Palmisano LJ, Silveira PA, Godfrey DI, Baxter AG. (1998) alpha/beta-T cell receptor (TCR)+CD4−CD8− (NKT) thymocytes prevent insulin-dependent diabetes mellitus in nonobese diabetic (NOD)/Lt mice by the influence of interleukin (IL)-4 and/or IL-10. J Exp Med 187:1047PubMedGoogle Scholar