Abstract 2303: Innate immunity driving tumor angiogenesis: the role of natural killer cells in non small cell lung cancer (NSCLC) .
Authors: Albini Adriana , Bruno Antonino , Pagani Arianna , Imperatori Andrea , Spagnoletti Marco , Rotolo Nicola , Cantelmo AnnaRita , Franzi Francesca , Capella Carlo , Ferlazzo Guido , Mortara Lorenzo , Noonan Douglas M. 
Published: August 2014

Abstract
Immune cells infiltrating tumors often show a skewed phenotype that reflects attenuation of anti-tumor activity and enhancement of pro-tumor activities, including angiogenesis.
Natural Killer (NK) cells are effectors lymphocytes of innate immunity that can potentially control tumors by their cytotoxic activity. Nevetherless the role of NK cells in angiogenesis remains to be defined.
Here we investigate the subset distribution of tumor infiltrating NKs and the correlation with pro-angiogenic factor production, like VEGF, PlGF and IL-8, on clinical samples belonging from patient with non-small cell lung cancer (NSCLC).
Samples (tumor, adjacent normal tissue and peripheral blood), solid tissues are mechanically dispersed, while blood is processed using a Ficoll gradient. Flow cytometry (FC) analyses were performed to evaluate specific markers (CD56, CD16, CD335, CD3) for NK cells.
The CD56brightCD16−NK phenotype predominated in all NSCLC samples while the CD56dimCD16+ cytotoxic NK phenotype prevailed in adjacent normal tissues and in non-oncologic lung tissues, independent of smoking status. The CD56+CD16− NK subset was associated with angiogenic cytokines production like VEGF, PlGF and IL-8 (CXCL8)
Further, patients with the SQK histotype showed significantly higher angiogenic factor production by CD56+CD16−NK cells infiltrating tumors, adjacent tissues and peripheral blood, compared to those from adenocarcinoma (ADK) and control tissues.
Supernatants derived from NSCLC infiltrating CD56+CD16−NK cells induced endothelial cell chemotaxis and formation of capillary-like structures in vitro, particularly evident in SQK derived NK cell supernatants and absent in controls
As TGFβ1 expression has been associated with poorer survival in non-small cell lung cancer (NSCLC) squamous cell carcinoma (SQK) and polarization of peripheral NK cells towards a decidual NK phenotype, a subset associated with angiogenesis, we evaluated whether TGFb culd be implicated in these polarization phenomena. We demonstrated as exposure to TGFβ1 up-regulated VEGF and PlGF production by peripheral blood CD56+CD16−NK cells from healthy subjects.
Our data suggest that, like other immune cells, NK cells in NSCLC can be switched to an angiogenic phenotype, particularly evident for SQK and likely mediated by TGFβ1, which could contribute to angiogenesis in SQK NSCLC development and progression.
Citation Format: Adriana Albini, Antonino Bruno, Arianna Pagani, Andrea Imperatori, Marco Spagnoletti, Nicola Rotolo, AnnaRita Cantelmo, Francesca Franzi, Carlo Capella, Guido Ferlazzo, Lorenzo Mortara, Douglas M. Noonan. Innate immunity driving tumor angiogenesis: the role of natural killer cells in non small cell lung cancer (NSCLC) . [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2303. doi:10.1158/1538-7445.AM2013-2303