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Rapid Detection of Neutrophil Oxidative Burst Capacity is Predictive of Whole Blood Cytokine Responses.

Vernon PJ, Schaub LJ, Dallelucca JJ, Pusateri AE, Sheppard FR - PLoS ONE (2015)

Bottom Line: Pearson's test was utilized to correlate NOX capacity with cytokine secretion, p<0.05 considered significant.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.These data suggest that NOX capacity may quickly identify patients at risk for maladaptive immune responses and who may benefit from immuno-modulatory therapies.

View Article: PubMed Central - PubMed

Affiliation: Naval Medical Research Unit San Antonio, JBSA-Ft. Sam Houston, Texas, United States of America.

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.

Conclusion: 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.

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Baseline neutrophil oxidative burst capacity is predictive of ex-vivo whole blood production of IL-1β, IL-6, TNFα, and the Th1-polarizing cytokine, IL-12/23(p40).(A) Scatter plots of cytokine fold change (y-axis) and neutrophil oxidative burst fold change in MFI (x-axis) that were determined to have statistically significant correlative coefficient (r) values (p≤0.0001) by Pearson’s Test. (B) Scatter plots of cytokine fold change and neutrophil oxidative burst capacity fold change for two cytokines found to be not statistically significant: IL-4 (Th2 cytokine), IL-17 (Th17 cytokine).
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pone.0146105.g003: Baseline neutrophil oxidative burst capacity is predictive of ex-vivo whole blood production of IL-1β, IL-6, TNFα, and the Th1-polarizing cytokine, IL-12/23(p40).(A) Scatter plots of cytokine fold change (y-axis) and neutrophil oxidative burst fold change in MFI (x-axis) that were determined to have statistically significant correlative coefficient (r) values (p≤0.0001) by Pearson’s Test. (B) Scatter plots of cytokine fold change and neutrophil oxidative burst capacity fold change for two cytokines found to be not statistically significant: IL-4 (Th2 cytokine), IL-17 (Th17 cytokine).

Mentions: Neutrophil oxidative burst capacity was determined to be positively correlated with the secretion of the cytokines G-CSF, GM-CSF, IL-1β, IL-1RA, IL-2, IL-6, IL-8, IL-12/23(p40), MIP-1α, and TNFα (p<0.05) (Table 1). For the purpose of determining the predictive ability of NOX capacity with regard to cytokine production, a much more stringent statistical threshold of p≤0.0001 was used. NOX capacity measured rapidly in fresh blood could predict the production of IL-1β (r = 0.6615, R2 = 0.4356), IL-6 (r = 0.7378, R2 = 0.5443) TNFα (r = 0.7564, R2 = 0.5722) and the Th1-polarizing cytokine IL-12/23(p40) (r = 0.7789, R2 = 0.6068) during a 24 hour ex-vivo stimulation with a high degree of accuracy (Fig 3A). Conversely, the Th2-associated cytokine IL-4 and the Th17-associated cytokine IL-17A were not correlated with NOX capacity with r values of 0.1689 and -0.167 and R2 values of 0.02853 and 0.02789, respectively (Fig 3B). These data suggest that in addition to the ability of NOX capacity to predict the extent of production of the broad pro-inflammatory markers IL1β, IL-6 and TNFα, which are generally present in elevated quantities during most tissue injury, it is also predictive of the specifically Th1-polarizing cytokine, IL-12/23(p40).


Rapid Detection of Neutrophil Oxidative Burst Capacity is Predictive of Whole Blood Cytokine Responses.

Vernon PJ, Schaub LJ, Dallelucca JJ, Pusateri AE, Sheppard FR - PLoS ONE (2015)

Baseline neutrophil oxidative burst capacity is predictive of ex-vivo whole blood production of IL-1β, IL-6, TNFα, and the Th1-polarizing cytokine, IL-12/23(p40).(A) Scatter plots of cytokine fold change (y-axis) and neutrophil oxidative burst fold change in MFI (x-axis) that were determined to have statistically significant correlative coefficient (r) values (p≤0.0001) by Pearson’s Test. (B) Scatter plots of cytokine fold change and neutrophil oxidative burst capacity fold change for two cytokines found to be not statistically significant: IL-4 (Th2 cytokine), IL-17 (Th17 cytokine).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4696850&req=5

pone.0146105.g003: Baseline neutrophil oxidative burst capacity is predictive of ex-vivo whole blood production of IL-1β, IL-6, TNFα, and the Th1-polarizing cytokine, IL-12/23(p40).(A) Scatter plots of cytokine fold change (y-axis) and neutrophil oxidative burst fold change in MFI (x-axis) that were determined to have statistically significant correlative coefficient (r) values (p≤0.0001) by Pearson’s Test. (B) Scatter plots of cytokine fold change and neutrophil oxidative burst capacity fold change for two cytokines found to be not statistically significant: IL-4 (Th2 cytokine), IL-17 (Th17 cytokine).
Mentions: Neutrophil oxidative burst capacity was determined to be positively correlated with the secretion of the cytokines G-CSF, GM-CSF, IL-1β, IL-1RA, IL-2, IL-6, IL-8, IL-12/23(p40), MIP-1α, and TNFα (p<0.05) (Table 1). For the purpose of determining the predictive ability of NOX capacity with regard to cytokine production, a much more stringent statistical threshold of p≤0.0001 was used. NOX capacity measured rapidly in fresh blood could predict the production of IL-1β (r = 0.6615, R2 = 0.4356), IL-6 (r = 0.7378, R2 = 0.5443) TNFα (r = 0.7564, R2 = 0.5722) and the Th1-polarizing cytokine IL-12/23(p40) (r = 0.7789, R2 = 0.6068) during a 24 hour ex-vivo stimulation with a high degree of accuracy (Fig 3A). Conversely, the Th2-associated cytokine IL-4 and the Th17-associated cytokine IL-17A were not correlated with NOX capacity with r values of 0.1689 and -0.167 and R2 values of 0.02853 and 0.02789, respectively (Fig 3B). These data suggest that in addition to the ability of NOX capacity to predict the extent of production of the broad pro-inflammatory markers IL1β, IL-6 and TNFα, which are generally present in elevated quantities during most tissue injury, it is also predictive of the specifically Th1-polarizing cytokine, IL-12/23(p40).

Bottom Line: Pearson's test was utilized to correlate NOX capacity with cytokine secretion, p<0.05 considered significant.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.These data suggest that NOX capacity may quickly identify patients at risk for maladaptive immune responses and who may benefit from immuno-modulatory therapies.

View Article: PubMed Central - PubMed

Affiliation: Naval Medical Research Unit San Antonio, JBSA-Ft. Sam Houston, Texas, United States of America.

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.

Conclusion: 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.

Show MeSH
Related in: MedlinePlus