The objective of this study was to characterize the effect of albumin therapy on the expression of Toll-like receptor 4 (TLR 4), anti-inflammation cytokines and CD4(+)CD25(+)forkhead box P3 (Foxp3)(+) regulatory T lymphocytes (Treg cells) in the ischemic brain and peripheral immune system after Middle Cerebral Artery Occlusion (MCAO). Adult male Kunming mice were subjected to MCAO, the suture was withdrawn 2 h later followed by an intravenous administration of 25% albumin (1.25 g/kg) or saline (5 ml/kg) through caudal vein. We demonstrated that albumin administration elevated the serum albumin level supranormally at 6 h and 24 h after MCAO in mice. In addition, we showed that both in the ischemic brain and in the spleen, albumin administration significantly depressed the increase of TLR 4 mRNA expression by quantitative real-time polymerase chain reaction (QRT-PCR), and significantly increase the anti-inflammatory related interleukin-10 (IL-10) and transforming growth factor beta1 (TGF-β1) mRNA expression by transcription-polymerase chain reaction (RT-PCR) after MCAO. In the spleen, compared to sham group, strong TLR 4 immunoreactivity was noted in the saline group; while compared to saline group, albumin administration markedly reduced the immunoreactivity of TLR 4 after MCAO by immunohistochemistry. Moreover, albumin administration significantly increased the percentage of Treg in spleen CD4(+) cells by flow cytometry. In conclusion, the decrease of TLR 4 expression and the increase of Treg cell, IL-10, and TGF-β1 expression may partly contribute to the neuroprotective effect of albumin therapy on MCAO induced immune inflammatory responses.