BACKGROUND: Nuclear factor-kappa B (NF-kappa B) plays a critical role in the cellular response to a variety of stimuli, and it regulates the production of various inflammatory cytokines, adhesion molecules, and enzymes. Polymorphonuclear leukocytes (PMNLs) play a central role in systemic inflammatory response after severe insult. The role of NF-kappa B in activation of PMNLs, however, has not been clear. We developed a simple flow cytometric method for quantifying expression of intranuclear NF-kappa B in PMNLs, and we used it to evaluate NF-kappa B activity in patients with systemic inflammatory response syndrome (SIRS). METHODS: Thirty patients who fulfilled the criteria for SIRS and 24 healthy volunteers were included as study subjects. Expression of intranuclear NF-kappa B with and without stimulation by lipopolysaccharide was quantified by our new method. Oxidative activity in PMNLs with and without formylmethionyl-leucyl-phenylalanine stimulation was measured by flow cytometry. Levels of interleukin-6, interleukin-8, PMNL elastase, and nitric oxide metabolites in blood were also measured. RESULTS: Expression of intranuclear NF-kappa B in PMNLs both with and without LPS stimulation was significantly elevated in SIRS patients in comparison with that of healthy volunteers. PMNL oxidative activity was significantly elevated in SIRS patients. Positive correlation was observed between intranuclear NF-kappa B expression and PMNL oxidative activity, whereas no relation was observed between intranuclear NF-kappa B expression and serum concentrations of chemical mediators. CONCLUSION: Our new flow cytometric method proved useful for quantifying intranuclear NF-kappa B expression in PMNLs. In PMNLs from SIRS patients, intranuclear NF-kappa B expression and oxidative activity were significantly elevated with positive correlation, and enhanced expression of NF-kappa B may play an important role in PMNL activation in SIRS.
BACKGROUND:Nuclear factor-kappa B (NF-kappa B) plays a critical role in the cellular response to a variety of stimuli, and it regulates the production of various inflammatory cytokines, adhesion molecules, and enzymes. Polymorphonuclear leukocytes (PMNLs) play a central role in systemic inflammatory response after severe insult. The role of NF-kappa B in activation of PMNLs, however, has not been clear. We developed a simple flow cytometric method for quantifying expression of intranuclear NF-kappa B in PMNLs, and we used it to evaluate NF-kappa B activity in patients with systemic inflammatory response syndrome (SIRS). METHODS: Thirty patients who fulfilled the criteria for SIRS and 24 healthy volunteers were included as study subjects. Expression of intranuclear NF-kappa B with and without stimulation by lipopolysaccharide was quantified by our new method. Oxidative activity in PMNLs with and without formylmethionyl-leucyl-phenylalanine stimulation was measured by flow cytometry. Levels of interleukin-6, interleukin-8, PMNL elastase, and nitric oxide metabolites in blood were also measured. RESULTS: Expression of intranuclear NF-kappa B in PMNLs both with and without LPS stimulation was significantly elevated in SIRS patients in comparison with that of healthy volunteers. PMNL oxidative activity was significantly elevated in SIRS patients. Positive correlation was observed between intranuclear NF-kappa B expression and PMNL oxidative activity, whereas no relation was observed between intranuclear NF-kappa B expression and serum concentrations of chemical mediators. CONCLUSION: Our new flow cytometric method proved useful for quantifying intranuclear NF-kappa B expression in PMNLs. In PMNLs from SIRS patients, intranuclear NF-kappa B expression and oxidative activity were significantly elevated with positive correlation, and enhanced expression of NF-kappa B may play an important role in PMNL activation in SIRS.
Authors: S M Lewis; D F Treacher; J Edgeworth; G Mahalingam; C S Brown; T A Mare; M Stacey; R Beale; K A Brown Journal: Clin Exp Immunol Date: 2015-09-22 Impact factor: 4.330
Authors: Maike E van Hezel; Margit Boshuizen; Anna L Peters; M Straat; Alexander P Vlaar; Angelique M E Spoelstra-de Man; Michael W T Tanck; Anton T J Tool; Boukje M Beuger; Taco W Kuijpers; Nicole P Juffermans; Robin van Bruggen Journal: Transfusion Date: 2019-12-05 Impact factor: 3.157