The purpose of this study was to examine the autonomic mechanisms underlying changes in heart rate (HR) and systolic blood pressure (SBP) responses to endotracheal tube (ETT) suctioning and to compare the open versus closed methods of ETT suctioning on these measures and on arterial oxygen tension. Eighteen orally intubated participants, 33 to 82 years of age (M = 60 years), were randomized for the order of suctioning method. Arterial oxygen tension (PaO2) was measured before suctioning and 30 s and 5 min following suctioning. Beat-to-beat HR and arterial blood pressure data were collected for 10-min periods before and after suctioning. HR and SBP measures were analyzed before suctioning and 1 min and 5 min following suctioning. Although there were no significant effects of ETT suctioning on the autonomic mechanisms of HR modulation and no significant differences between the two methods of suctioning, ETT suctioning resulted in an increase in HR, SBP, and PaO2. However, there was a decrease in the parasympathetic nervous system indicator of HR variability (HRV) following open suctioning. All patients in this study maintained a PaO2 level 80 mm Hg, which may account for our lack of significant autonomic changes. This suggests that hyperoxygenation with 100% oxygen for a minimum of 1 min (or 20 breaths), as delivered by preoxygenation modes available on most microprocessor ventilators, should be the method of choice for all hyperoxygenation procedures to avoid a decrease in PaO2 following suctioning.