High PCO(2) levels attenuate reperfusion injury and ventilation-induced injury in isolated and perfused lungs. We asked whether premature lambs could tolerate 6 h of ventilation with a PCO(2) >80 mm Hg and whether the high PCO(2) modulated the ventilator-induced injury. Preterm surfactant-treated lambs were ventilated for 30 min with a high tidal volume (V(T)) to induce lung injury. The lambs then were ventilated for 5.5 h with a V(T) of 6-9 mL/kg to achieve a PCO(2) of 40-50 mm Hg in the control group. CO(2) was added to the ventilator circuit of a high PCO(2) group to maintain an average PCO(2) of 95 +/- 5 mm Hg. The high PCO(2) lambs had heart rates, blood pressures, plasma cortisol values, and oxygenation equivalent to the control lambs. The lungs of the high PCO(2) group had significantly higher gas volumes and had less lung injury by histopathology. Indicators of inflammation (white blood cells, hydrogen peroxide production, and IL-1beta and IL-8 cytokine mRNA expression in cells from the alveolar wash) qualitatively indicated less injury in the high PCO(2) group, although the differences were not significant. Preterm lambs tolerated a very high PCO(2) without physiologic compromise for 6 h. The high PCO(2) may attenuate ventilator-induced lung injury in the preterm.