Impaired neutrophil chemotaxis in sepsis associates with GRK expression and inhibition of actin assembly and tyrosine phosphorylation

• Published in: Blood Vol. 108; no. 9; pp. 2906 - 2913
• Main Authors: Arraes, Sandra Mara A., Freitas, Marta S., da Silva, Simone V., de Paula Neto, Heitor A., Alves-Filho, Jose Carlos, Martins, Maria Auxiliadora, Basile-Filho, Anibal, Tavares-Murta, Beatriz M., Barja-Fidalgo, Christina, Cunha, Fernando Q.
• Format: Journal Article
• Language: English
• Published: Washington, DC Elsevier Inc 01.11.2006; The Americain Society of Hematology
• Subjects: Actins - biosynthesis; beta-Adrenergic Receptor Kinases - genetics; Biological and medical sciences; Cell Movement; Chemotaxis, Leukocyte - physiology; Cytokines - pharmacology; DNA Primers; Fundamental and applied biological sciences. Psychology; Fundamental immunology; G-Protein-Coupled Receptor Kinase 2; G-Protein-Coupled Receptor Kinase 5; Humans; Immunobiology; Lipopolysaccharides - pharmacology; Myeloid cells: ontogeny, maturation, markers, receptors; N-Formylmethionine Leucyl-Phenylalanine - pharmacology; Neutrophils - drug effects; Neutrophils - physiology; Phosphotyrosine - metabolism; Polynuclears; Protein-Serine-Threonine Kinases - genetics; Receptors, Interleukin-8A - genetics; Reverse Transcriptase Polymerase Chain Reaction; Sepsis - blood; Human; Phosphorylation; Enzyme; Transferases; Granulocyte; Inflammation; Chemotaxis; Infection; Sepsis syndrome; G protein-coupled receptor kinase; Leukotriene B4; Actin; Neutrophil; Protein-tyrosine kinase; Tumor necrosis factor α; Interleukin 8; Gamma interferon
• ISSN: 0006-4971; 1528-0020
• DOI: 10.1182/blood-2006-05-024638

The deregulation of inflammatory response during sepsis seems to reflect the overproduction of mediators, which suppress leukocyte functions. We investigated the intracellular mechanisms underlying the inability of neutrophils from severe septic patients to migrate toward chemoattractants. Patients with sepsis (52) and 15 volunteers were prospectively enrolled. Patients presented increased circulating levels of tumor necrosis factor-α, interferon-γ, interleukin (IL)–8, and IL-10. Patients showed reduced neutrophil chemotaxis to formyl-methionyl-leucyl-phenylalanine (FMLP), leukotriene B4 (LTB4) or IL-8. No difference in the transcription or expression of the IL-8 receptor, CXCR1, was detected in neutrophils from controls and patients. However, septic neutrophils failed to increase tyrosine phosphorylation and actin polymerization in response to IL-8 or LTB4. In contrast, septic neutrophils, similar to controls, showed phagocytic activity that induced actin polymerization and augmented phosphotyrosine content. Treatment of control neutrophils with cytokines and lipopolysaccharide (LPS) to mimic endogenous septic environment inhibited actin polymerization and tyrosine phosphorylation in response to IL-8 or LTB4. High expression of G protein–coupled receptor kinase 2 (GRK2) and GRK5 was detected in septic neutrophils and control cells treated with cytokines plus LPS. Data suggest that endogenous mediators produced during sepsis might continually activate circulating neutrophils, leading to GRK activation, which may induce neutrophil desensitization to chemoattractants.