Tumor necrosis factor- (TNF) is associated with sleep regulation in health and disease. Previous studies assessed sleep in mice genetically deficient in the TNF 55 kD receptor. In this study, spontaneous and influenza virus-induced sleep profiles were assessed in mice deficient in both the 55 kD and 75 kD TNF receptors (TNF-2R knockouts [KO]) and wild-type (WT) strain controls. Under baseline conditions the TNF-2R KO mice had less non-rapid-eye-movement sleep (NREMS) than WTs during the nighttime and more rapid-eye-movement sleep (REMS) than controls during the daytime. The differences between nighttime maximum and daytime minimum values of electroencephalogram (EEG) delta power during NREMS were greater in the TNF-2R KO mice than in WTs. Viral challenge (mouse-adapted influenza X-31) enhanced NREMS and decreased REMS in both strains roughly to the same extent. EEG delta power responses to viral challenge differed substantially between strains; the WT animals increased, whereas the TNF-2R KO mice decreased their EEG delta wave power during NREMS. There were no differences between strains in body temperatures or locomotor activity in uninfected mice or after viral challenge. Analyses of cortical mRNAs confirmed that the TNF-2R KO mice lacked both TNF receptors; these mice also had higher levels of orexin mRNA and reduced levels of the purine P2X7 receptor compared to WTs. Results reinforce the hypothesis that TNF is involved in physiological sleep regulation but plays a limited role in the acute phase response induced by influenza virus.