title: Extracellular acidification induces human neutrophil activation.
abstract: In the current work, we evaluated the effect of extracellular acidification on neutrophil physiology. Neutrophils suspended in bicarbonate-buffered RPMI 1640 medium adjusted to acidic pH values (pH 6.5-7.0) underwent: 1) a rapid transient increase in intracellular free calcium concentration levels; 2) an increase in the forward light scattering properties; and 3) the up-regulation of surface expression of CD18. By contrast, extracellular acidosis was unable to induce neither the production of H2O2 nor the release of myeloperoxidase. Acidic extracellular pH also modulated the functional profile of neutrophils in response to conventional agonists such as FMLP, precipiting immune complexes, and opsonized zymosan. It was found that not only calcium mobilization, shape change response, and up-regulation of CD18 expression but also production of H2O2 and release of myeloperoxidase were markedly enhanced in neutrophils stimulated in acidic pH medium. Moreover, extracellular acidosis significantly delayed neutrophil apoptosis and concomitantly extended neutrophil functional lifespan. Extracellular acidification induced an immediate and abrupt fall in the intracellular pH, which persisted over the 240-s analyzed. A similar abrupt drop in the intracellular pH was detected in cells suspended in bicarbonate-supplemented PBS but not in those suspended in bicarbonate-free PBS. A role for intracellular acidification in neutrophil activation is suggested by the fact that only neutrophils suspended in bicarbonate-buffered media (i.e., RPMI 1640 and bicarbonate-supplemented PBS) underwent significant shape changes in response to extracellular acidification. Together, our results support the notion that extracellular acidosis may intensify acute inflammatory responses by inducing neutrophil activation as well as by delaying spontaneous apoptosis and extending neutrophil functional lifespan.
authors: Laboratory of Immunology, Institute of Hematologic Research, National Academy of Medicine, Buenos Aires University School of Medicine, Argentina. neu@connmed.com.ar
authors: A S
authors: AS
authors: Trevani
authors: G
authors: G
authors: Andonegui
authors: M
authors: M
authors: Giordano
authors: D H
authors: DH
authors: López
authors: R
authors: R
authors: Gamberale
authors: F
authors: F
authors: Minucci
authors: J R
authors: JR
authors: Geffner
mesh_terms: D000936:Antigen-Antibody Complex / Q000494:pharmacology; D018821:CD18 Antigens / Q000096:biosynthesis / Q000097:blood; D020013:Calcium Signaling / Q000276:immunology; D048429:Cell Size / Q000276:immunology; D002470:Cell Survival / Q000276:immunology; D003593:Cytoplasm / Q000378:metabolism; D005110:Extracellular Space / Q000276:immunology / Q000378:metabolism* / Q000502:physiology; D006801:Humans; D006851:Hydrochloric Acid; D006861:Hydrogen Peroxide / Q000097:blood / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007424:Intracellular Fluid / Q000276:immunology / Q000378:metabolism; D007552:Isotonic Solutions; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D018375:Neutrophil Activation / Q000187:drug effects / Q000276:immunology; D009195:Peroxidase / Q000097:blood / Q000378:metabolism; D015054:Zymosan / Q000494:pharmacology
chemical_list: D000936:Antigen-Antibody Complex; D018821:CD18 Antigens; D007552:Isotonic Solutions; D009240:N-Formylmethionine Leucyl-Phenylalanine; D015054:Zymosan; D006861:Hydrogen Peroxide; D009195:Peroxidase; D006851:Hydrochloric Acid