Abstract. V · 24NKT cells are lymphocytes expressing both T-cell antigen receptors and NK-cell antigen receptors on their cell surface and are involved in tumor immunity. They exert their antitumor effects after being activated by a specific ligand, ·-galactosyl ceramide (·-GalCer). Malignant pleural effusion, a frequently occurring complication in patients with lung cancer, contains numerous lymphocytes. In the present study, we examined the presence and functions of V · 24NKT cells in the lymphocytes in pleural effusion in vitro. The subjects were 13 untreated patients with primary lung cancer, who suffered malignant pleural effusion as a complication and who were treated between April 2004 and October 2007 at our hospital. Mononuclear cells were separated from the malignant pleural effusion and incubated with ·-GalCer and IL-12. The production of IFN-Á and IL-4 after incubation and the proportion of the V · 24NKT cells before and after incubation were determined and compared. In the group cultured with ·-GalCer alone, no significant increase in IFN-Á production was observed in comparison with the control group. In the group cultured with ·-GalCer + IL-12, IFN-Á production increased significantly in comparison with the control group, and the proportion of V · 24NKT cells increased after incubation. IL-4 production was very much lower than IFN-Á production. V · 24NKT cells were present in malignant pleural effusion in patients with lung cancer, but IFN-Á production did not increase after addition of ·-GalCer alone. The V · 24NKT cells were activated by ·-GalCer in the presence of IL-12. The V · 24NKT cells in malignant pleural effusion were not activated by ·-GalCer alone, suggesting that V · 24NKT cell function is attenuated in malignant pleural effusion.