People living with HIV (PLWH) have increasingly longer life spans. This age group faces different challenges than younger PLWH, which may include increased stress and social isolation. The purpose of this study was to determine if the age and sex of PLWH is associated with measures of physiologic stress, perceived stress and social isolation. In this cross-sectional study, we enrolled 102 PLWH equally into four groups divided by age (< or > 50 years) and gender. Participants completed well-validated survey measurements of stress and isolation, and their heart rate variability over 60 minutes was measured by Holter monitor. The mean (standard deviation) Perceived Stress Scale score was 17.4 (6.94), mean visual analog stress scale score was 3.51 (2.79), and mean Hawthorne Friendship Scale score, a measure of social isolation, was 17.03 (4.84). Mean heart rate variability expressed as the standard deviation of successive N-N intervals was 65.47 (31.16) msec. In multivariable regression models that controlled for selected demographic variables, there was no relationship between the Perceived Stress Scale and age (coefficient=−0.09, p=−.23) or female gender (coefficient=−0.12, p=0.93); however, there was a modest relationship between female gender and stress using the visual analog stress scale (coefficient=1.24, p=0.05). Perceived Stress was negatively associated with the Hawthorne Friendship score (coefficient=−0.34, p=0.05). Hawthorne Friendship score was positively associated with younger age (coefficient=0.11, p=0.02). Age was the only independent predictor of physiologic stress as measured by heart rate variability (coefficient=−1.3, p<0.01). Our findings suggest that younger PLWH may experience more social isolation; however, age-related changes in heart rate variability do not appear to be related to perceived stress or social isolation. Future longitudinal research is required to more thoroughly understand this relationship and its impact on the health of PLWH.