OBJECTIVES: To determine whether blood neutrophils from healthy individuals and blood and synovial fluid neutrophils from patients with rheumatoid arthritis (RA) responded differently to priming agonists and stimuli of the oxidative burst and, if so, whether this was a property of a subpopulation of neutrophils. METHODS: Continuous flow electrophoresis was used to separate neutrophils into subpopulations based upon quantitative differences in net negative surface charge. The generation of superoxide anion (O2-) was used as a measure of oxidative activity using 10(-7) mol/l N-formyl-methionylleucyl-phenylalanine (FMLP) as the stimulating agonist and 10(-8) mol/l platelet activating factor (PAF) as the priming agent. RESULTS: The production of O2- by blood and synovial fluid neutrophils from RA patients in response to FMLP was greater than that observed with control blood neutrophils (p < 0.001). Priming of normal blood neutrophils with PAF increased their FMLP induced oxidative burst (p < 0.001), but PAF treatment had no effect on rheumatoid neutrophils. Neutrophils from synovial fluid of RA patients were less electronegative than paired blood samples and exposure of blood neutrophils to FMLP but not PAF reduced their surface charge. Continuous flow electrophoresis isolated three neutrophil subpopulations: cells of least surface electronegativity were ascribed to pool P1 and cells of greatest surface electro-negativity to P3. Normal blood neutrophils from P3, but not P1, showed increased oxidative activity after PAF priming (twofold increase; p < 0.01), whereas the responsiveness of rheumatoid blood and synovial fluid neutrophils from P1 and P3 was not modified by PAF treatment under the same conditions. CONCLUSION: It is suggested that most of the circulating neutrophils in RA are already in a state of readiness to generate O2- upon activation by an inflammatory stimulus. This is in contrast to normal blood neutrophils, which have both responsive and non-responsive subpopulations with respect to priming agonists.
OBJECTIVES: To determine whether blood neutrophils from healthy individuals and blood and synovial fluid neutrophils from patients with rheumatoid arthritis (RA) responded differently to priming agonists and stimuli of the oxidative burst and, if so, whether this was a property of a subpopulation of neutrophils. METHODS: Continuous flow electrophoresis was used to separate neutrophils into subpopulations based upon quantitative differences in net negative surface charge. The generation of superoxide anion (O2-) was used as a measure of oxidative activity using 10(-7) mol/l N-formyl-methionylleucyl-phenylalanine (FMLP) as the stimulating agonist and 10(-8) mol/l platelet activating factor (PAF) as the priming agent. RESULTS: The production of O2- by blood and synovial fluid neutrophils from RApatients in response to FMLP was greater than that observed with control blood neutrophils (p < 0.001). Priming of normal blood neutrophils with PAF increased their FMLP induced oxidative burst (p < 0.001), but PAF treatment had no effect on rheumatoid neutrophils. Neutrophils from synovial fluid of RApatients were less electronegative than paired blood samples and exposure of blood neutrophils to FMLP but not PAF reduced their surface charge. Continuous flow electrophoresis isolated three neutrophil subpopulations: cells of least surface electronegativity were ascribed to pool P1 and cells of greatest surface electro-negativity to P3. Normal blood neutrophils from P3, but not P1, showed increased oxidative activity after PAF priming (twofold increase; p < 0.01), whereas the responsiveness of rheumatoid blood and synovial fluid neutrophils from P1 and P3 was not modified by PAF treatment under the same conditions. CONCLUSION: It is suggested that most of the circulating neutrophils in RA are already in a state of readiness to generate O2- upon activation by an inflammatory stimulus. This is in contrast to normal blood neutrophils, which have both responsive and non-responsive subpopulations with respect to priming agonists.
Authors: L E Leigh; B Ghebrehiwet; T P Perera; I N Bird; P Strong; U Kishore; K B Reid; P Eggleton Journal: Biochem J Date: 1998-02-15 Impact factor: 3.857
Authors: U Kishore; R D Sontheimer; K N Sastry; K S Zaner; E G Zappi; G R Hughes; M A Khamashta; P Strong; K B Reid; P Eggleton Journal: Biochem J Date: 1997-03-01 Impact factor: 3.857
Authors: Jean Lucas G da Silva; Daniela F Passos; Viviane M Bernardes; Fernanda L Cabral; Paulo G Schimites; Alessandra G Manzoni; Edilene Gadelha de Oliveira; Cristiane de Bona da Silva; Ruy Carlos Ruver Beck; Matheus H Jantsch; Roberto M Maciel; Daniela B R Leal Journal: Inflammation Date: 2019-10 Impact factor: 4.092
Authors: Otavio Cabral-Marques; Tabata Takahashi França; Ashraf Al-Sbiei; Lena Friederike Schimke; Taj Ali Khan; Claudia Feriotti; Tania Alves da Costa; Osvaldo Reis Junior; Cristina Worm Weber; Janaíra Fernandes Ferreira; Fabiola Scancetti Tavares; Claudia Valente; Regina Sumiko Watanabe Di Gesu; Asif Iqbal; Gabriela Riemekasten; Gustavo Pessini Amarante-Mendes; José Alexandre Marzagão Barbuto; Beatriz Tavares Costa-Carvalho; Paulo Vitor Soeiro Pereira; Maria J Fernandez-Cabezudo; Vera Lucia Garcia Calich; Luigi D Notarangelo; Troy R Torgerson; Basel K Al-Ramadi; Hans D Ochs; Antonio Condino-Neto Journal: J Allergy Clin Immunol Date: 2018-03-05 Impact factor: 10.793