Supplemental Figure 2. (A) Gating strategy for NKG2D expression on NK cells in FNA and PBMCs from an ATC patient. Lymphocytes are gated according to size (forward scatter FSC) and granularity (side scatter SSC), viability (7AAD- cells), NK cell phenotype (CD3-CD56+ cells) and NKG2D expression is plotted as a histogram where solid grey histogram represent isotype control antibody staining and open histogram represent staining with NKG2D-specific antibody. (B) Gating strategy for ULBP2/5/6 expression in FNA from an ATC patient. Tumor cells are first gated according to size (forward scatter FSC), granularity (side scatter SSC) and viability (LIVE/DEAD- cells). Granulocytes that may appear in the same FSC/SSC population are gated out by excluding the CD11c+ cells. ULBP2/5/6 expression is plotted as a histogram where solid grey histogram represents isotype control. antibody staining and open histogram represents staining with ULBP2/5/6-specific antibody. FNA = fine needle aspirate. PBMC = peripheral blood mononuclear cells.
ARTICLE ABSTRACT
Purpose: Anaplastic thyroid carcinoma (ATC) is one of the most aggressive forms of cancer with no curative therapies available. To date, strategies to target ATC by immunotherapy have not been evaluated. We investigated whether ATC would be a suitable target for natural killer (NK) cell–based immunotherapy.Experimental Design: We first established seven new cell lines from ATC tumors, three from papillary thyroid carcinoma tumors and analyzed them together with eight additional ATC cell lines. Cells were analyzed for sensitivity to lysis by NK cells and their ability to chemoattract and regulate the activity of NK cells. In addition, fresh tumor samples and peripheral blood from six patients with ATC were analyzed for NK cell infiltration and phenotype.Results: We observed that ATC cell lines are sensitive to lysis by ex vivo expanded NK cells and that the lysis was abrogated upon blockade of NKG2D. Sensitivity of thyroid cancer cell lines to NK cell–mediated lysis correlated with surface expression of UL16-binding protein 2 on tumor cells. Moreover, ATC cell lines produced high levels of CXCL10 and stimulated migration of expanded NK cells and ATC tumors were enriched for NK cells expressing the cognate chemokine receptor CXCR3. However, compared with NK cells in peripheral blood, ATC tumor–derived NK cells displayed a suppressed phenotype with a downregulated expression of NKG2D. In vitro, suppression of NK cell–mediated lysis and NKG2D expression by ATC cells was restored upon neutralization of prostaglandin-E2.Conclusions: ATC cell lines are sensitive to NK cell–mediated lysis via ULBP2/5/6 and chemoattract CXCR3-positive NK cells. Patients with ATC may benefit from NK cell–based immunotherapy. Clin Cancer Res; 20(22); 5733–44. ©2014 AACR.
