Objective. To investigate the kinetic and reversibility of mechanical ventilation-associated pulmonary and systemic inflammatory response in patients with acute lung injury (ALI). Design. Prospective observational cross-over study. Setting. Intensive care unit of a university hospital. Patients. Twelve mechanically ventilated patients with ALI. Interventions. Mechanical ventilation was transiently changed from a lung protective setting with PEEP of 15cmH2O and a VT of 5ml/kg predicted body weight to a more conventional ventilatory setting with PEEP of 5 cmH2O and VT of 12ml/kg predicted body weight for a period of 6h. Measurements and results. We examined the profile of interleukin (IL)-1, IL-1 receptor antagonist, IL-6, IL-10, and tumor necrosis factor in the plasma of all patients, and in the bronchoalveolar lavage (mini-BAL) fluid of six of these patients. Measurements were performed at baseline, 1h, and 6h after each change of the ventilatory setting. Switching to conventional mechanical ventilation was associated with a higher PaO2 (P 0.05) and a marked increase (P 0.05) of measured plasma cytokines in patients with and without mini-BAL with a maximum after 1h. Similarly, intraalveolar cytokine concentrations increased with conventional mechanical ventilation. While plasma cytokine levels returned to baseline values, intraalveolar cytokine concentrations further increased when lung protective mechanical ventilation was reestablished. Conclusions. In patients with ALI, initiation of low PEEP and high VT mechanical ventilation is associated with cytokine release into circulation which occurred within 1h. It is independent from BAL procedures and can be reversed by reinstitution of lung protective mechanical ventilation.