Alveolar macrophages (m phi) participate in inflammatory and immune re
sponses in acidic microenvironments such as the interstitial fluids of
tumors and abscesses. Two plasmalemmal H+ extruders interact to contr
ol the acid-base status of alveolar m phi, namely a V-type H+ pump (V-
ATPase) and a Na+/H+ exchanger, The present study examined the effects
of extracellular pH (pH(o)) and H+ transport inhibitors on tumor necr
osis factor-alpha (TNF-alpha) release induced by endotoxin (lipopolysa
ccharide) in rabbit alveolar m phi. The amount and activity of TNF-alp
ha in m phi-conditioned media were determined by enzyme-linked immunos
orbent assay and L929 fibroblast bioassay, respectively. TNF-alpha rel
ease was suppressed progressively at lower pH(o), values (less than or
equal to 7.0). Also, bafilomycin A(l) (a specific inhibitor of V-ATPa
ses) significantly reduced the amount and activity of TNF-alpha in m p
hi-conditioned media (pH(o) 7.4). However, bafilomycin caused a signif
icant increase in the nonspecific cytotoxicity (i.e. bioactivity insen
sitive to TNF-alpha antibody) of m phi-conditioned media. The effects
of bafilomycin specifically on TNF-alpha release followed a time cours
e similar to that of acidic pH(o), suggesting that both treatments act
ed on similar events in the lipopolysaccharide signal transduction pat
hway. Amiloride (an inhibitor of Na+ transporters including the Na+/H exchanger) also suppressed TNF-alpha release but displayed a time cou
rse of action different from the acidic pH(o) or bafilomycin.