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. Author manuscript; available in PMC: 2013 Aug 1.
Published in final edited form as: Cancer Res. 2012 May 30;72(15):3744–3752. doi: 10.1158/0008-5472.CAN-11-2695

Figure 1. Tumor Ascites Inhibits CD1d-mediated NKT cell activation.

Figure 1

(A) LCD1dwt cells were treated with control medium or with ovarian cancer ascites fluid from patients for 4 h, then washed extensively and cocultured with a panel of NKT cell hybridomas (DN32.D3, N38-3C3 and N37-1A12). After 20–24 h, IL-2 was measured as an indication of NKT cell activation using standard cytokine ELISA. (B) The effect of ascites on NKT cell recognition of CD1d1 molecules is rapid. LCD1d1wt cells were incubated with ascites for the indicated times. The cells were then co-cultured with NKT cell hybridomas and IL-2 release was measured. (C) Conditioned medium from ascites-derived cells is inhibitory. LCD1d1wt cells were incubated for 4 h with fresh cell culture medium, ascites fluid, or conditioned medium from ascites-derived cells from patient OC-95. Treated APCs were then co-cultured with NKT cell hybridomas and IL-2 release was measured. Data are shown as mean ±S.E.M. of one experiment set up in triplicate. The experiments were performed at least twice with each ascites sample.