Human neutrophil stimulation by influenza virus: relationship of cytoplasmic pH changes to cell activation.
We have previously demonstrated that influenza A virus (IAV) stimulates the human neutrophil through phospholipase C activation. With the use of the fluorescent indicator 2',7'-bis(carboxyethyl)-5,6-carboxyfluorescein (BCECF), cytoplasmic acidification and subsequent alkalinization are shown to accompany this activation. These responses are not inhibited by pertussis toxin (PT). The alkalinization is mediated largely *but not entirely) by the Na(+)-H+ antiporter and is not initiated, or modulated, by the IAV-induced cytosolic Ca2+ (Cai2+) rise. Rather, protein kinase C (PKC) is likely the mediator of cell alkalinization, based on studies using the PKC inhibitor 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H-7). The acidification can be dissociated from the alkalinization response, which is also independent of Cai2+ fluxes and of PKC. Both pHi responses can be dissociated from the respiratory burst. Cytosolic alkalinization and acidification seem to reflect two independently mediated responses of the activated neutrophil, the former resulting ultimately from phospholipase activation and the latter from other activities that are not yet fully characterized.
Hartshorn
K L
KL
William B, Castle Hematology Research Laboratory, Boston City Hospital, Massachusetts.
Wright
J
J
Collamer
M A
MA
White
M R
MR
Tauber
A I
AI
D002118:Calcium / Q000097:blood; D002772:Cholera Toxin / Q000494:pharmacology; D003593:Cytoplasm / Q000502:physiology; D004533:Egtazic Acid / Q000494:pharmacology / Q000031:analogs & derivatives; D006801:Humans; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D009980:Influenza A virus / Q000502:physiology*; D007700:Kinetics; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000187:drug effects / Q000502:physiology*; D037342:Pertussis Toxin; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
D010566:Virulence Factors, Bordetella; D004533:Egtazic Acid; D009240:N-Formylmethionine Leucyl-Phenylalanine; D002772:Cholera Toxin; D037342:Pertussis Toxin; C025603:1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium