RESUMO
An important checkpoint in the progression of melanoma is the metastasis to lymph nodes. Here, to investigate the role of lymph node NK cells in disease progression, we analyze frequency, phenotype and functions of NK cells from tumour-infiltrated (TILN) and tumour-free ipsilateral lymph nodes (TFLN) of the same patients. We show an expansion of CD56(dim)CD57(dim)CD69 + CCR7 +KIR+ NK cells in TILN. TILN NK cells display robust cytotoxic activity against autologous melanoma cells. In the blood of metastatic melanoma patients, the frequency of NK cells expressing the receptors for CXCL8 receptor is increased compared with healthy subjects, and blood NK cells also express the receptors for CCL2 and IL-6. These factors are produced in high amount in TILN and in vitro switch the phenotype of blood NK cells from healthy donors to the phenotype associated with TILN. Our data suggest that the microenvironment of TILN generates and/or recruits a particularly effective NK cell subset.
Assuntos
Células Matadoras Naturais/imunologia , Linfonodos/imunologia , Melanoma/imunologia , Neoplasias Cutâneas/imunologia , Adulto , Idoso , Antígenos CD/metabolismo , Antígenos de Diferenciação de Linfócitos T/metabolismo , Antígeno CD56/metabolismo , Antígenos CD57/metabolismo , Feminino , Humanos , Células Matadoras Naturais/metabolismo , Lectinas Tipo C/metabolismo , Linfonodos/patologia , Metástase Linfática , Masculino , Melanoma/patologia , Pessoa de Meia-Idade , Receptores CCR7/metabolismo , Receptores de Interleucina-6/metabolismo , Receptores KIR2DL2/metabolismo , Receptores KIR2DL3/metabolismo , Neoplasias Cutâneas/patologia , Microambiente Tumoral/imunologiaRESUMO
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.
Assuntos
Antígenos de Diferenciação de Linfócitos T/metabolismo , Células Matadoras Naturais/imunologia , Melanoma/imunologia , Receptores Desencadeadores da Citotoxicidade Natural/metabolismo , Animais , Anticorpos/imunologia , Anticorpos/farmacologia , Antígenos de Diferenciação de Linfócitos T/imunologia , Antígenos Ly/genética , Antígenos Ly/metabolismo , Linhagem Celular Tumoral , Testes Imunológicos de Citotoxicidade , Citotoxicidade Imunológica/efeitos dos fármacos , Citotoxicidade Imunológica/imunologia , Proteínas de Ligação a DNA/genética , Humanos , Imunoterapia Adotiva/métodos , Subunidade gama Comum de Receptores de Interleucina/genética , Células Matadoras Naturais/metabolismo , Células Matadoras Naturais/transplante , Ligantes , Metástase Linfática/imunologia , Antígeno-1 Associado à Função Linfocitária/metabolismo , Melanoma/metabolismo , Melanoma/patologia , Melanoma/terapia , Melanoma Experimental/metabolismo , Camundongos , Camundongos Endogâmicos NOD , Camundongos Endogâmicos , Camundongos Knockout , Camundongos SCID , Camundongos Transgênicos , Receptor 1 Desencadeador da Citotoxicidade Natural/genética , Receptor 1 Desencadeador da Citotoxicidade Natural/metabolismo , Células T Matadoras Naturais/imunologia , Receptores Desencadeadores da Citotoxicidade Natural/imunologia , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Early hematopoietic zinc finger/zinc finger protein 521 (EHZF/ZNF521) is a novel zinc finger protein expressed in hematopoietic stem and progenitor cells and is down-regulated during their differentiation. Its transcript is also abundant in some hematopoietic malignancies. Analysis of the changes in the antigenic profile of cells transfected with EHZF cDNA revealed up-regulation of HLA class I cell surface expression. This phenotypic change was associated with an increased level of HLA class I H chain, in absence of detectable changes in the expression of other Ag-processing machinery components. Enhanced resistance of target cells to NK cell-mediated cytotoxicity was induced by enforced expression of EHZF in the cervical carcinoma cell line HeLa and in the B lymphoblastoid cell line IM9. Preincubation of transfected cells with HLA class I Ag-specific mAb restored target cell susceptibility to NK cell-mediated lysis, indicating a specific role for HLA class I Ag up-regulation in the NK resistance induced by EHZF. A potential clinical significance of these findings is further suggested by the inverse correlation between EHZF and MHC class I expression levels, and autologous NK susceptibility of freshly explanted multiple myeloma cells.