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David J. Hackam, MD; Sergio Grinstein, PhD; Avery Nathens, MD; R. William G. Watson, PhD; John C. Marshall, MD; Ori D. Rotstein, MD

Arch Surg. 1996;131(12):1296-1301.

Abstract
Objective
To investigate the effects of neutrophil (PMN) transmigration into inflammatory sites on cytosolic pH (pH_i) regulation.

Design
Exudative PMNs were obtained by bronchoalveolar lavage from rats sustaining endotoxin-induced lung injury. Circulating PMNs were purified with density-gradient centrifugation. Cytosolic pH was measured with single-cell fluorescence imaging using the pH-sensitive dye biscarboxyethyl-carboxyfluorescein.

Results
Exudative PMNs showed impaired pH, recovery from an induced acid load compared with circulating PMNs. Under conditions of extracellular acidosis, exudative PMNs showed impaired pH_i homeostasis and produced decreased superoxide compared with circulating cells. Inhibition of the sodiumproton exchanger attenuated the differences in pH_i recovery, suggesting a mechanism underlying the pH_i regulatory dysfunction. All cells had comparable adenosine triphosphate levels and superoxide production at physiologic extracellular pH.

Conclusion
Impaired pH, regulation of exudative cells may mediate cellular dysfunction and impaired resolution of infection at inflammatory sites.

Arch Surg. 1996;131:1296-1301

Author Affiliations
From the Department of Surgery, The Toronto Hospital, University of Toronto (Drs Hackam, Nathens, Watson, Marshall, and Rotstein), and the Department of Cell Biology, Hospital for Sick Children (Dr Grinstein), Toronto, Ontario.

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