Objective A potential anti‐inflammatory role for acetylcholinesterase inhibitors (AChEIs) has been supported by animal studies. As very limited data exist from individuals with Alzheimer's disease (AD), the aim of this study was to assess the potential influence of AChEIs on blood pro‐inflammatory cytokines. We hypothesized that pro‐inflammatory cytokine concentrations were lower in individuals with AD stabilized on AChEIs. Methods Blood interleukin‐1 beta, interleukin‐6, and tumor necrosis factor‐alpha concentrations were assessed using specific enzyme‐linked immunosorbent assays in three groups of participants: patients with AD stabilized on a therapeutic dose of an AChEI (n = 42); AChEIs drug naïve patients (n = 24); and a cognitively unimpaired control group (n = 35). Patients in the AChEIs group had received medication for an average of one year. Results Patients stabilized on an AChEI did not differ significantly from drug naïve patients in relation to the concentrations of interleukin‐1 beta, interleukin‐6, and tumor necrosis factor‐alpha (p = 0.874, 0.225, and 0.978, respectively). Within the group taking AChEIs, the levels of cytokines did not differ between those taking donepezil, rivastigmine, or galantamine (p = 0.368, 0.851, and 0.299, respectively). Conclusions Results from animal studies suggesting a modulatory anti‐inflammatory role for AChEIs was not advanced in this study. In individuals with AD, very limited evidence currently exists to support the hypothesis that AChEIs may influence inflammatory blood markers and function beyond the enhancement of neuronal transmission. However, further studies assessing a wider range of inflammatory markers and processes are still needed before this hypothesis can be ruled out. Copyright © 2013 John Wiley & Sons, Ltd.