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. Author manuscript; available in PMC: 2017 Aug 15.
Published in final edited form as: Clin Cancer Res. 2016 Apr 13;22(16):4249–4258. doi: 10.1158/1078-0432.CCR-15-2518

Figure 1. Tumor Ascites and Conditioned medium from ovarian cancer cell lines inhibits NKT cell activation.

Figure 1

(A) LCD1d 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, N37-1A12, N38-3C3 and N38-2C12). After 20-24 h, IL-2 was measured as an indication of NKT cell activation using standard cytokine ELISA. (B, C) LCD1d cells were treated with supernatants from confluent ovarian cancer cell lines OVCAR-3 and SK-OV-3 for 4 hours at 37°C and washed extensively following treatment. Control LCD1d cells were concurrently treated with RPMI (B+ Medium) and McCoy media. Lvector cells serve as a negative control. N.D.=not detectable. Following treatment, LCD1d cells were co-cultured with NKT cell hybridomas, DN32.D3, and incubated for 24 hours at 37° C. Standard ELISA was performed to measure cytokine production (B) IL-2 (C) IL-4. 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 and three times with conditioned medium. T-tests were performed to compare medium vs. conditioned medium groups, yielding p-value of p=0.0061 for IL-2 and p=0.0187 for IL-4.