Neutrophil Extracellular Traps in Plasma from Dogs with Immune-mediated Hemolytic Anemia

C. Lawson, S. A. Smith, M. O'Brien, M. McMichael

Research output: Contribution to journalArticlepeer-review


Background: Neutrophil extracellular traps (NETs) are part of the innate immune response and are essential in local pathogen control, but are associated with pathological inflammation, organ damage, autoimmunity, and thrombosis. Immune-mediated hemolytic anemia (IMHA) is a pro-inflammatory, prothrombotic disease associated with high mortality. Hypothesis/Objectives: Neutrophil extracellular traps (NETs) are a feature of the inflammatory process in dogs with IMHA. The objective of the study was to evaluate plasma from dogs with IMHA for the presence of 2 indirect markers and 1 direct marker of NETs. Animals: Healthy client-owned dogs (56) and hospitalized dogs with IMHA (n = 35). Methods: Prospective study. Plasma samples for all dogs were evaluated for cell-free DNA using a fluorescence assay, histone-DNA (hisDNA) complex using an ELISA, and citrullinated histone H3 (specific for NETosis) using Western blot. Reference intervals were generated using plasma from healthy dogs. Results: In dogs with IMHA, cell-free DNA concentration was above the reference interval in 17% of samples with a median (range) of 1.0 μg/mL (0.1–17.3), and hisDNA concentration was above the reference interval in 94% of samples with a median (range) of 30.7 × pooled normal plasma (PNP; 0.6–372.1). Western blot for citrullinated histone H3 identified detectable bands in 84% samples from dogs with IMHA. Conclusions and Clinical Importance: The assay for cell-free DNA detected evidence of NETs in fewer dogs than did the other approaches. Excessive NETs appears to be a feature of IMHA in dogs and contributions to the prothrombotic state deserve further study.

Original languageEnglish (US)
Pages (from-to)128-134
Number of pages7
JournalJournal of veterinary internal medicine
Issue number1
StatePublished - Jan 1 2018


  • DNA
  • Histone
  • IMHA
  • Inflammation

ASJC Scopus subject areas

  • General Veterinary


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