title: Nicotine drives neutrophil extracellular traps formation and accelerates collagen-induced arthritis.
abstract: The aim was to investigate the effects of nicotine on neutrophil extracellular traps (NETs) formation in current and non-smokers and on a murine model of RA.
  
  
  
  We compared spontaneous and phorbol 12-myristate 13-acetate-induced NETosis between current and non-smokers by DNA release binding. Nicotine-induced NETosis from non-smokers was assessed by DNA release binding, NET-specific (myeloperoxidase (MPO)-DNA complex) ELISA and real-time fluorescence microscopy. We also used immunofluorescent staining to detect nicotinic acetylcholine receptors (nAChRs) on neutrophils and performed a functional analysis to assess the role of nAChRs in nicotine-induced NETosis. Finally, we investigated the effects of systemic nicotine exposure on arthritis severity and NETosis in the CIA mouse model.
  
  
  
  Neutrophils derived from current smokers displayed elevated levels of spontaneous and phorbol 12-myristate 13-acetate-induced NETosis. Nicotine induced dose-dependent NETosis in ex vivo neutrophils from healthy non-smokers, and co-incubation with ACPA-immune complexes or TNF-α facilitated a synergistic effect on NETosis. Real-time fluorescence microscopy revealed robust formation of NET-like structures in nicotine-exposed neutrophils. Immunofluorescent staining demonstrated the presence of the α7 subunit of the nAChR on neutrophils. Stimulation of neutrophils with an α7-specific nAChR agonist induced NETosis, whereas pretreatment with an nAChR antagonist attenuated nicotine-induced NETosis. Nicotine administration to mice with CIA exacerbated inflammatory arthritis, with higher plasma levels of NET-associated MPO-DNA complex.
  
  
  
  We demonstrate that nicotine is a potent inducer of NETosis, which may play an important role in accelerating arthritis in the CIA model. This study generates awareness of and the mechanisms by which nicotine-containing products, including e-cigarettes, may have deleterious effects on patients with RA.
authors.affiliation: Division of Immunology and Rheumatology, Stanford University School of Medicine, Stanford, CA, USA.|Department of Medicine, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.|Department of Medicine, VA Palo Alto Healthcare System, Palo Alto.|Department of Internal Medicine, Stanford University School of Medicine, Stanford, CA.
authors.forename: Jaejoon
authors.initials: J
authors.lastname: Lee
authors.affiliation: Division of Immunology and Rheumatology, Stanford University School of Medicine, Stanford, CA, USA.|Department of Medicine, VA Palo Alto Healthcare System, Palo Alto.|Department of Internal Medicine, Stanford University School of Medicine, Stanford, CA.
authors.forename: Ayala
authors.initials: A
authors.lastname: Luria
authors.affiliation: Division of Immunology and Rheumatology, Stanford University School of Medicine, Stanford, CA, USA.|Department of Medicine, VA Palo Alto Healthcare System, Palo Alto.|Department of Internal Medicine, Stanford University School of Medicine, Stanford, CA.
authors.forename: Christopher
authors.initials: C
authors.lastname: Rhodes
authors.affiliation: Division of Immunology and Rheumatology, Stanford University School of Medicine, Stanford, CA, USA.|Department of Medicine, VA Palo Alto Healthcare System, Palo Alto.|Department of Internal Medicine, Stanford University School of Medicine, Stanford, CA.
authors.forename: Harini
authors.initials: H
authors.lastname: Raghu
authors.affiliation: Division of Immunology and Rheumatology, Stanford University School of Medicine, Stanford, CA, USA.|Department of Medicine, VA Palo Alto Healthcare System, Palo Alto.|Department of Internal Medicine, Stanford University School of Medicine, Stanford, CA.
authors.forename: Nithya
authors.initials: N
authors.lastname: Lingampalli
authors.affiliation: Division of Immunology and Rheumatology, Stanford University School of Medicine, Stanford, CA, USA.|Department of Medicine, VA Palo Alto Healthcare System, Palo Alto.|Department of Internal Medicine, Stanford University School of Medicine, Stanford, CA.
authors.forename: Orr
authors.initials: O
authors.lastname: Sharpe
authors.affiliation: Department of Infectious Diseases, University of Georgia College of Veterinary Medicine, Athens, GA, USA.
authors.forename: Balazs
authors.initials: B
authors.lastname: Rada
authors.affiliation: Department of Microbiology and Immunology, Pusan National University School of Medicine, Pusan, Korea.
authors.forename: Dong Hyun
authors.initials: DH
authors.lastname: Sohn
authors.affiliation: Division of Immunology and Rheumatology, Stanford University School of Medicine, Stanford, CA, USA.|Department of Medicine, VA Palo Alto Healthcare System, Palo Alto.|Department of Internal Medicine, Stanford University School of Medicine, Stanford, CA.
authors.forename: William H
authors.initials: WH
authors.lastname: Robinson
authors.affiliation: Division of Immunology and Rheumatology, Stanford University School of Medicine, Stanford, CA, USA.|Department of Medicine, VA Palo Alto Healthcare System, Palo Alto.|Department of Internal Medicine, Stanford University School of Medicine, Stanford, CA.
authors.forename: Jeremy
authors.initials: J
authors.lastname: Sokolove
mesh_terms: D000818:Animals; D001169:Arthritis, Experimental / Q000209:etiology; D001172:Arthritis, Rheumatoid / Q000209:etiology*; D002356:Cartilage / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D066300:Electronic Nicotine Delivery Systems; D004797:Enzyme-Linked Immunosorbent Assay; D065206:Extracellular Traps / Q000187:drug effects*; D006801:Humans; D055104:Infusions, Subcutaneous; D008297:Male; D008811:Mice, Inbred DBA; D009504:Neutrophils / Q000187:drug effects; D009538:Nicotine / Q000008:administration & dosage / Q000494:pharmacology*; D018722:Nicotinic Agonists / Q000008:administration & dosage / Q000494:pharmacology*; D009195:Peroxidase / Q000378:metabolism; D012907:Smoking / Q000009:adverse effects; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
chemical_list: D018722:Nicotinic Agonists; D009538:Nicotine; D009195:Peroxidase; D013755:Tetradecanoylphorbol Acetate
keywords: animal model; collagen-induced arthritis; neutrophil extracellular traps; nicotine; rheumatoid arthritis