Mature Cytotoxic CD56bright/CD16+Natural Killer Cells Can Infiltrate Lymph Nodes Adjacent to Metastatic Melanoma

Meriem Messaoudene,Isabelle Cremer,Eve Maubec,Sebastien Albert,Emmanuelle Fourmentraux-Neves,Xavier Sastre-Garau,Nicolas Jacquelot,Johan Chanal,Delphine Mitilian,Sarra Mazouz-Dorval,Benoit Couturaud,Charles Guédon,Marie-Francoise Avril,Anne Caignard,Giulia Fregni,Laurence Zitvogel,Angelique Girod,Lydia Deschamps,Sylvie Rusakiewicz

doi:10.1158/0008-5472.can-13-1303

Abstract

Melanomas are characterized by high metastatic potential, with regional lymph node representing the most frequent site of early dissemination in this disease. These regional lymph nodes also represent the primary site for differentiation of natural killer (NK) cells. Although blood-derived NK cells can efficiently lyse melanoma cells isolated from metastatic lymph node (M-LN), there has been no study of the properties of the most disease-relevant NK cells isolated from M-LN in patients with melanoma. Here, we report that M-LN contains 0.5% to 11% of CD56(bright) NK cells among CD45(+) hematopoietic cells present and that this cell population surrounds tumor cell clusters in M-LN. This NK cell population was characterized by expression of CD62L, chemokine receptors, and high levels of natural cytotoxicity receptors (NCR), NK group 2 D (NKG2D), and DNAX accessory molecule 1 (DNAM-1). Expression of NCR-NKp30 and NKG2D correlated negatively with percentages of tumor cells in M-LN. Interestingly, M-LN contained a unique subset of mature CD56(bright)CD16(+) NK cells displaying coregulated expression of NCR and NKG2D activating receptors. Ex vivo analyses suggested that M-LN-derived NK cells were inactive but could be activated by appropriate cytokine signals [interleukin (IL)-2 or IL-15], and could lyse metastatic melanoma cells in a highly efficient manner compared with blood-derived NK cells. Taken together, the results offer evidence that adjuvant immunotherapy that targets NK cells in M-LN for activation may improve treatment of patients with sentinel lymph node-positive melanoma.

Highlights

Melanomas are highly metastatic tumors for which the treatment of advanced stages is still unsatisfactory
In metastatic lymph node (M-LN), the lymph node architecture was altered by invading tumor cells: natural killer (NK) cells were distributed around the lymphoid structures, squeezed under the lymph node capsule, and rarely detected in the tumor cell clusters (Fig. 1A)
Phenotypic analyses by multiparametric flow cytometry were performed in 22 M-LN obtained from 16 stage III patients with melanoma treated by lymph node dissection (Table 1) and in four of six D-LN

Summary

Introduction

Melanomas are highly metastatic tumors for which the treatment of advanced stages is still unsatisfactory. The main way of dissemination is the lymph and the invasion of sentinel lymph node, the initial tumor-draining lymph node (LN), is correlated with prognosis. Stage III patients are characterized by lymph node metastases in the draining basin. These patients are treated by lymph node. Authors' Affiliations: 1INSERM U1016, CNRS UMR 8104, Institut Cochin; 2APHP, Department of Dermatology, Hospital Cochin, University Paris Descartes; 3APHP, Department of Dermatology; 4APHP, Department of Surgery, Hospital Bichat, University Paris Diderot; 5Surgery Department, Institut Curie, rue d'Ulm-Paris; 6Hospital Foch, Suresnes; 7Centre de Recherche des Cordeliers, 15 rue de l'ecole de medecine; and 8INSERM U1015 INSERM, Institut Gustave Roussy, Villejuif, Paris, France. Fregni: Institute of Pathology, Centre Hospitalier Universitaire Vaudois, Faculty of Biology and Medicine, University of Lausanne, Lausanne, Switzerland

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