NCRs and DNAM-1 mediate NK cell recognition and lysis of human and mouse melanoma cell lines in vitro and in vivo
Tadepally Lakshmikanth, Shannon Burke, Talib Hassan Ali, Silvia Kimpfler, Francesco Ursini, Loredana Ruggeri, Marusca Capanni, Viktor Umansky, Annette Paschen, Antje Sucker, Daniela Pende, Veronika Groh, Roberto Biassoni, Petter Höglund, Masashi Kato, Kazuko Shibuya, Dirk Schadendorf, Andrea Anichini, Soldano Ferrone, Andrea Velardi, Klas Kärre, Akira Shibuya, Ennio Carbone, Francesco Colucci
Tadepally Lakshmikanth, Shannon Burke, Talib Hassan Ali, Silvia Kimpfler, Francesco Ursini, Loredana Ruggeri, Marusca Capanni, Viktor Umansky, Annette Paschen, Antje Sucker, Daniela Pende, Veronika Groh, Roberto Biassoni, Petter Höglund, Masashi Kato, Kazuko Shibuya, Dirk Schadendorf, Andrea Anichini, Soldano Ferrone, Andrea Velardi, Klas Kärre, Akira Shibuya, Ennio Carbone, Francesco Colucci
View: Text | PDF
Research Article Oncology

NCRs and DNAM-1 mediate NK cell recognition and lysis of human and mouse melanoma cell lines in vitro and in vivo

Abstract

NK cells use a variety of receptors to detect abnormal cells, including tumors and their metastases. However, in the case of melanoma, it remains to be determined what specific molecular interactions are involved and whether NK cells control metastatic progression and/or the route of dissemination. Here we show that human melanoma cell lines derived from LN metastases express ligands for natural cytotoxicity receptors (NCRs) and DNAX accessory molecule-1 (DNAM-1), two emerging NK cell receptors key for cancer cell recognition, but not NK group 2 member D (NKG2D). Compared with cell lines derived from metastases taken from other anatomical sites, LN metastases were more susceptible to NK cell lysis and preferentially targeted by adoptively transferred NK cells in a xenogeneic model of cell therapy. In mice, DNAM-1 and NCR ligands were also found on spontaneous melanomas and melanoma cell lines. Interference with DNAM-1 and NCRs by antibody blockade or genetic disruption reduced killing of melanoma cells. Taken together, these results show that DNAM-1 and NCRs are critical for NK cell–mediated innate immunity to melanoma cells and provide a background to design NK cell–based immunotherapeutic strategies against melanoma and possibly other tumors.

Authors

Tadepally Lakshmikanth, Shannon Burke, Talib Hassan Ali, Silvia Kimpfler, Francesco Ursini, Loredana Ruggeri, Marusca Capanni, Viktor Umansky, Annette Paschen, Antje Sucker, Daniela Pende, Veronika Groh, Roberto Biassoni, Petter Höglund, Masashi Kato, Kazuko Shibuya, Dirk Schadendorf, Andrea Anichini, Soldano Ferrone, Andrea Velardi, Klas Kärre, Akira Shibuya, Ennio Carbone, Francesco Colucci

×

Figure 6

Differential contribution of NKp46 and DNAM-1.

Options: View larger image (or click on image) Download as PowerPoint
Differential contribution of NKp46 and DNAM-1. (A) CFSE-labeled RET cell...
(A) CFSE-labeled RET cells (2 × 105) were injected i.p. into control IgG– and DNAM-1–treated or NK1.1-depleted B6 mice, and the number of residual RET cells were counted 48 hours later. Representative plots of peritoneal lavage CFSE gating are shown. (B) Quantification of residual cells. Data are mean + SEM of 8–10 mice per group and are pooled from 2 experiments. *P < 0.05 compared with control. (C) NK cells were expanded for 5 days in IL-2, sorted into DNAM-1+ and DNAM-1–, recultured for 48 hours, and tested for cytotoxicity at the indicated E:T ratio. Results are representative of 2–3 experiments. (D) NK cells from WT or Ncr1–/– mice were expanded, sorted, recultured as described in C, and injected i.p. into Rag2–/–Il2rg–/– mice (2.5 × 105/mouse) immediately after i.p. injection of 5 × 105 CFSE-labeled RET cells. Residual RET cells were counted following lavage 48 hours later. Results are individual mice from 3 independent experiments. The bar represents the mean of 8–9 mice per group. (E and F) RET cells (105) were injected i.v. into control IgG, anti–DNAM-1–treated or anti-NK1.1–depleted WT or Ncr1–/– mice (E). (F) RET cells (2 × 105) were injected i.v. into WT, Ncr1–/–, or NK1.1-depleted WT mice. Lung metastases were counted 14 days later. Results are from individual mice, and the bar represents the mean of 4–5 (E) and 6–11 (F) mice per group. **P < 0.01.