title: Rapid Detection of Neutrophil Oxidative Burst Capacity is Predictive of Whole Blood Cytokine Responses.
abstract: BACKGROUND
  Maladaptive immune responses, particularly cytokine and chemokine-driven, are a significant contributor to the deleterious inflammation present in many types of injury and infection. Widely available applications to rapidly assess individual inflammatory capacity could permit identification of patients at risk for exacerbated immune responses and guide therapy. Here we evaluate neutrophil oxidative burst (NOX) capacity measured by plate reader to immuno-type Rhesus Macaques as an acute strategy to rapidly detect inflammatory capacity and predict maladaptive immune responses as assayed by cytokine array.
  
  
  METHODS
  Whole blood was collected from anesthetized Rhesus Macaques (n = 25) and analyzed for plasma cytokine secretion (23-plex Luminex assay) and NOX capacity. For cytokine secretion, paired samples were either unstimulated or ex-vivo lipopolysaccharide (LPS)-stimulated (100μg/mL/24h). NOX capacity was measured in dihydrorhodamine-123 loaded samples following phorbol 12-myristate 13-acetate (PMA)/ionomycin treatment. Pearson's test was utilized to correlate NOX capacity with cytokine secretion, p<0.05 considered significant.
  
  
  RESULTS
  LPS stimulation induced secretion of the inflammatory molecules G-CSF, IL-1β, IL-1RA, IL-6, IL-10, IL-12/23(p40), IL-18, MIP-1α, MIP-1β, and TNFα. Although values were variable, several cytokines correlated with NOX capacity, p-values≤0.0001. Specifically, IL-1β (r = 0.66), IL-6 (r = 0.74), the Th1-polarizing cytokine IL-12/23(p40) (r = 0.78), and TNFα (r = 0.76) were strongly associated with NOX.
  
  
  CONCLUSIONS
  NOX capacity correlated with Th1-polarizing cytokine secretion, indicating its ability to rapidly predict inflammatory responses. These data suggest that NOX capacity may quickly identify patients at risk for maladaptive immune responses and who may benefit from immuno-modulatory therapies. Future studies will assess the in-vivo predictive value of NOX in animal models of immune-mediated pathologies.
authors.affiliation: Naval Medical Research Unit San Antonio, JBSA-Ft. Sam Houston, Texas, United States of America.
authors.forename: Philip J
authors.initials: PJ
authors.lastname: Vernon
authors.affiliation: Naval Medical Research Unit San Antonio, JBSA-Ft. Sam Houston, Texas, United States of America.
authors.forename: Leasha J
authors.initials: LJ
authors.lastname: Schaub
authors.affiliation: 59th Medical Wing, US Air Force, JBSA-Ft. Sam Houston, Texas, United States of America.
authors.forename: Jurandir J
authors.initials: JJ
authors.lastname: Dallelucca
authors.affiliation: US Army Medical Research and Materiel Command, Ft. Detrick, Maryland, United States of America.
authors.forename: Anthony E
authors.initials: AE
authors.lastname: Pusateri
authors.affiliation: Naval Medical Research Unit San Antonio, JBSA-Ft. Sam Houston, Texas, United States of America.
authors.forename: Forest R
authors.initials: FR
authors.lastname: Sheppard
mesh_terms: D000818:Animals; D054405:Chemokine CCL3 / Q000097:blood; D054407:Chemokine CCL4 / Q000097:blood; D016207:Cytokines / Q000097:blood* / Q000502:physiology; D005434:Flow Cytometry; D016179:Granulocyte Colony-Stimulating Factor / Q000097:blood; D007249:Inflammation / Q000503:physiopathology; D018664:Interleukin-12 / Q000097:blood; D020382:Interleukin-18 / Q000097:blood; D053583:Interleukin-1beta / Q000097:blood; D015850:Interleukin-6 / Q000097:blood; D008253:Macaca mulatta; D009504:Neutrophils / Q000502:physiology*; D016897:Respiratory Burst / Q000502:physiology*; D014409:Tumor Necrosis Factor-alpha / Q000097:blood
chemical_list: D054405:Chemokine CCL3; D054407:Chemokine CCL4; D016207:Cytokines; D020382:Interleukin-18; D053583:Interleukin-1beta; D015850:Interleukin-6; D014409:Tumor Necrosis Factor-alpha; D016179:Granulocyte Colony-Stimulating Factor; D018664:Interleukin-12