Carbon-based nanomaterials including carbon nanotubes (CNTs) have been shown to trigger inflammation. However, how these materials are 'sensed' by immune cells is not known. Here we compared the effects of two carbon-based nanomaterials, single-walled CNTs (SWCNTs) and graphene oxide (GO), on primary human monocyte-derived macrophages. Genome-wide transcriptomics assessment was performed at sub-cytotoxic doses. Pathway analysis of the microarray data revealed pronounced effects on chemokine-encoding genes in macrophages exposed to SWCNTs, but not in response to GO, and these results were validated by multiplex array-based cytokine and chemokine profiling. Conditioned medium from SWCNT-exposed cells acted as a chemoattractant for dendritic cells. Chemokine secretion was reduced upon inhibition of NF-κB, as predicted by upstream regulator analysis of the transcriptomics data, and Toll-like receptors (TLRs) and their adaptor molecule, MyD88 were shown to be important for CCL5 secretion. Moreover, a specific role for TLR2/4 was confirmed by using reporter cell lines. Computational studies to elucidate how SWCNTs may interact with TLR4 in the absence of a protein corona suggested that binding is guided mainly by hydrophobic interactions. Taken together, these results imply that CNTs may be 'sensed' as pathogens by immune cells.

Department of Chemical Sciences and Technologies University of Rome Tor Vergata Rome 00133 Italy

Department of Experimental Medicine and Surgery University of Rome Tor Vergata Rome 00173 Italy

Department of Genetic Toxicology and Nanotoxicology Institute of Experimental Medicine AS CR 14220 Prague Czech Republic

Department Pharmacology and Physiology West Virginia University Morgantown WV 26505 USA

Exposure Assessment Branch National Institute for Occupational Safety and Health Morgantown WV 26505 USA

Faculty of Social Sciences University of Tampere 33014 Tampere Finland

Inorganic and Materials Chemistry University of Cologne 50939 Cologne Germany

Laboratory of Cellular Immunology Humanitas Clinical and Research Institute 20089 Rozzano Milano Italy

Laboratory of Environmental Toxicology National Institute of Chemical Physics and Biophysics Tallinn 12618 Estonia

Nanosafety and Nanomedicine Laboratory Division of Molecular Toxicology Institute of Environmental Medicine Karolinska Institutet 17177 Stockholm Sweden

Sanford Burnham Prebys Medical Discovery Institute La Jolla CA 92037 USA

Turku Centre for Biotechnology University of Turku 20520 Turku and Åbo Akademi University 20500 Turku Finland

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Immunotoxicity of Carbon-Based Nanomaterials, Starring Phagocytes