The aim of this cross-sectional study was to investigate associations of heart rate variability (HRV) with inflammatory markers among Buffalo, NY police officers. A total of 383 officers had complete data on HRV (high (HF) and low (LF) frequency power and heart rate) and inflammatory markers (C-reactive protein (CRP), interleukin 6 (IL-6), tumor necrosis factor-alpha (TNF-alpha) and fibrinogen). Electrocardiographic (ECG) data were processed using consensus standards for analysis of HRV; 5 minutes of resting ECG data were analyzed. Inflammatory markers were measured after fasting 12 hours using standard techniques. Linear regression and analysis of variance and covariance were used to assess mean levels of inflammatory markers across tertiles of HRV components. Univariate analysis revealed that HF and LF measures were strongly and inversely correlated with CRP (r = -0.21, P < 0.001 and r = -0.23, P < 0.001, respectively). In multivariate models, this relationship was attenuated and no longer significant (beta = -0.09, P = 0.057 and beta = -0.11, P = 0.066, respectively). Mean levels of TNF-alpha and fibrinogen decreased significantly with increasing tertiles of HF power (P = 0.004 and P = 0.028, respectively), but after multivariate adjustment for other risk factors results were attenuated (P = 0.097 and P = 0.992). Heart rate was positively and significantly associated with CRP and IL-6 across all models. Findings from this study were consistent with other studies where measures of vagal nerve activity, such as HF and LF power, were inversely related to inflammatory markers. Results suggest that cardiovascular risk factors account for some of the inverse association between HRV and inflammatory markers.