Altered intracellular pH regulation in neutrophils from patients with cystic fibrosis.
Cystic fibrosis (CF) is a condition characterized by neutrophil-mediated lung damage and bacterial colonization. The physiological basis for reported functional alterations in CF neutrophils, including increased release of neutrophil elastase, myeloperoxidase, and oxidants, is unknown. These processes are, however, regulated by intracellular pH (pH(i)). We demonstrate here that pH(i) regulation is altered in neutrophils from CF patients. Although resting pH(i) is similar, pH(i) after acid loading and activation (N-formyl-methionyl-leucyl-phenylalanine and phorbol 12-myristate 13-acetate) is more acidic in CF cells than in normal cells. Furthermore, patients with non-CF-related bronchiectasis handle acid loading and activation in a fashion similar to subjects with normal neutrophils, suggesting that chronic pulmonary inflammation alone does not explain the difference in pH(i). This is further supported by data showing that normal neutrophils exposed to the CF pulmonary milieu respond by increasing pH(i) as opposed to decreasing pH(i) as seen in activated CF neutrophils. These pH(i) differences in activated or acid-loaded CF neutrophils are abrogated by ZnCl(2) but not by amiloride and bafilomycin A(1), suggesting that passive proton conductance is abnormal in CF. In addition, DIDS, which inhibits HCO(3)(-)/Cl(-) exchange, causes alkalinization of control but not of CF neutrophils, suggesting that anion transport is also abnormal in CF neutrophils. In summary, we have shown that pH(i) regulation in CF neutrophils is intrinsically abnormal, potentially contributing to the pulmonary manifestations of the condition.
Coakley
R J
RJ
Pulmonary Research Division, Beaumont Hospital, Dublin 9, Ireland.
Taggart
C
C
Canny
G
G
Greally
P
P
O'Neill
S J
SJ
McElvaney
N G
NG
D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid / Q000494:pharmacology; D000143:Acids / Q000494:pharmacology; D000293:Adolescent; D000328:Adult; D000468:Alkalies / Q000378:metabolism; D001987:Bronchiectasis / Q000378:metabolism / Q000473:pathology; D001992:Bronchoalveolar Lavage Fluid / Q000166:cytology; D002712:Chlorides / Q000494:pharmacology; D003550:Cystic Fibrosis / Q000378:metabolism* / Q000473:pathology; D005260:Female; D006801:Humans; D006859:Hydrogen / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007425:Intracellular Membranes / Q000378:metabolism*; D008297:Male; D009000:Monocytes / Q000502:physiology; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism*; D011522:Protons; D012016:Reference Values; D017967:Zinc Compounds / Q000494:pharmacology
D000143:Acids; D000468:Alkalies; D002712:Chlorides; D011522:Protons; D017967:Zinc Compounds; D006859:Hydrogen; C016837:zinc chloride; D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid