Priming of Neutrophils and Differentiated PLB-985 Cells by Pathophysiological Concentrations of TNF-α Is Partially Oxygen Dependent

• Published in: Journal of innate immunity 2011-01, Vol.3 (3), p.298-314
• Main Authors: Volk, A. Paige Davis, Barber, Brieanna M., Goss, Kelli L., Ruff, Jake G., Heise, Christine K., Hook, Jessica S., Moreland, Jessica G.
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; CD11b Antigen - genetics; CD11b Antigen - metabolism; Cell Differentiation; Cell Line, Tumor; Cell Movement - immunology; Enzyme Activation - immunology; Fundamental and applied biological sciences. Psychology; Fundamental immunology; General aspects; Genetics of the immune response; Humans; Immunobiology; MAP Kinase Signaling System - immunology; Mathematics in biology. Statistical analysis. Models. Metrology. Data processing in biology (general aspects); Membrane Glycoproteins - genetics; Membrane Glycoproteins - metabolism; NADP - genetics; NADP - metabolism; NADPH Oxidase 2; NADPH Oxidases - genetics; NADPH Oxidases - metabolism; Neutrophil Activation; Neutrophils - immunology; Neutrophils - metabolism; Neutrophils - pathology; Oxygen - immunology; Oxygen - metabolism; p38 Mitogen-Activated Protein Kinases - metabolism; Research Article; Sepsis - immunology; Sepsis - prevention & control; Tumor Necrosis Factor-alpha - immunology; Tumor Necrosis Factor-alpha - metabolism
• ISSN: 1662-811X; 1662-8128
• DOI: 10.1159/000321439

Activation of polymorphonuclear leukocytes (PMN) can be modulated to intermediate ‘primed’ states characterized by enhanced responsiveness to subsequent stimuli. We studied priming in response to TNF-α in human PMN and PLB-985 cells, a myeloid cell line differentiated to a neutrophilic phenotype (PLB-D). PMN generated reactive oxygen species (ROS) in response to TNF-α alone, and NADPH oxidase activity increased in response to stimulation with formyl-Met-Leu-Phe after priming. PLB-D cells also demonstrated priming of NADPH oxidase activity. Similar to priming by endotoxin, priming of the respiratory burst by TNF-α was predominantly oxygen dependent, with marked attenuation of ROS generation if primed anaerobically. Both PMN and PLB-D cells displayed significant increases in cell surface CD11b and gp91 phox expression after TNF-α priming and PMN displayed activation of MAPK. In response to TNF-α priming, neither mobilization of intracellular proteins nor activation of MAPK pathways was NADPH oxidase dependent. Priming of PMN and PLB-D cells by low TNF-α concentrations enhanced chemotaxis. These data demonstrate that pathophysiological concentrations of TNF-α elicit NADPH oxidase-derived ROS and prime cells for enhanced surface protein expression, activation of p38 and ERK1/2 MAPK pathways, and increased chemotaxis. Furthermore, PLB-D cells undergo TNF-α priming and provide a genetically modifiable model to study priming mechanisms.