Figure 8.

Coculture of ex vivo isolated PDAC cells with autologous PBMC or TIL inhibits γδ T cell proliferation and enhances release of galectin-3. (A) In total, 5 × 10³ or (B) 1.5 × 10⁵ ex vivo isolated tumor cells (TuC) and (A) 2.5 × 10⁵ autologous PBMC or (B) 1.5 × 10³ autologous TIL were cultured alone or together in complete medium or were stimulated with 2.5 μM zoledronic acid with 50 IU/mL rIL-2 (Z). At day 0 and 11 days after culture, the absolute cell number of the Vγ9 γδ T cells was determined using SCDA. Cell culture supernatants were collected after 96 h and galectin-3 was determined by ELISA. Each symbol presents the data of one donor, and the lines represent the median of 4 different independent experiments. (A) Statistical comparison of matched samples was carried out parametrically by using paired, two-tailed t-test. P-value; *P < 0.05, ***P < 0.001. (B) Wilcoxon non-parametric, matched-pairs signed rank test (left panel) or parametric, matched-pairs, two-tailed t-test (right panel) was carried out. Samples present no significant differences. (C) Histograms are showing intracellular gal-3 expression stained with anti-gal-3 Ab (gray) compared to the appropriate isotype-control (unfilled) in pan-Cytokeratin⁺ tumor cells and CD45⁺ leukocytes of 2 representative donors (PDAC #1 and #2) out of 4.