The brain infarction volume was determined following a 72-h reperfusion period. signals were colocalized and significantly greater in mind cells adjacent to infracted mind lesions when compared to the corresponding regions of the contralateral hemisphere and to control (sham) mice. Treatment with IVIG and C1-INH efficiently reduced deposition of C3b and downregulated excessive TLR2 and p-JNK1 manifestation at the site of I/R injury. Taken together, these results provide a rationale for potential use of IVIG and C1-INH, alone or in combination with ischemic stroke and additional neurological conditions that involve inappropriately triggered components of the innate immune system. test, Chi-square and ANOVA followed by NewmanCKeuls post hoc checks for pairwise comparisons. Results IVIG is definitely Neuroprotective Both Before and After Ischemia in Experimental Stroke The effect of IVIG (Privigen, CSL Behring) on the outcome of mind I/R injury was evaluated in the murine stroke induced by 1 h MCAO and 72 h reperfusion. In non-treated (I/R only) mice, this procedure resulted in an average mind infarction of 36% of the affected hemisphere. IVIG treatment at 1 g/kg 30 min before and 1 h after MCAO as well as 3 h and 6 h post-ischemia at 2 g/kg reduced the infarction size by 56, 80, 58 and 59%, respectively. Treatment with control reagents, 250 mM l-proline (vehicle or stabilizer for immunoglobulin molecules in the Privigen IVIG preparation) or human being SRI-011381 hydrochloride albumin for IV use at 1 g/kg in vehicle solution experienced no significant effect on the size SRI-011381 hydrochloride of infarction when compared to non-treated mice (Fig. 1a). Open in a separate windows Fig. 1 IVIG exerts doseCresponsive neuroprotective effect in the mouse model of stroke when given before or after ischemia. a Mind infarction size 72 h post-ischemia in albumin treated (= 19), IVIG (1 g/kg 30 min before ischemia) pretreated (= 12), IVIG treated 1 h (Post SRI-011381 hydrochloride 1 h, = 30) and 3 h (Post 3 h, = 11) after ischemia compared with non-treated (I/R only, = 35) and vehicle (= 18) treated mice; ideals are mean SEM; * 0.001 b Mind infarct area in mice treated with reducing doses of IVIG at 1 g/kg (= 30), 0.5 g/kg (= 10) and 0.25 g/kg (= TNF-alpha 9); # 0.001, * 0.05. c Mortality in mice subjected to decreasing doses of IVIG at 1 g/kg (= 30), 0.5 g/kg (= 10) and 0.25 g/kg (= 9); d neurological deficit score evaluated at 72 h post-ischemia in mice treated with IVIG 1 g/kg (= 35), 0.5 g/kg (= 12) and 0.25 g/kg (= 13) To determine if the marked reduction of ischemic brain injury observed in IVIG-treated mice resulted in a reduced functional impairment, neurological deficits were evaluated daily at 24, 48 and 72 h in mice that were eventually killed for the brain infarction measurements. IVIG treatments significantly reduced the neurological deficits at SRI-011381 hydrochloride 48 and 72 h after the ischemia (Table 1). IVIG also exerted a dose-dependent effect on the consequences of ischemic mind injury. With half-decreasing doses of Privigen (1.0 g/kg, 0.5 g/kg and 0.25 g/kg) stepwise raises in mind infarction size (Fig. 1b), mortality (Fig. 1c) and neurological deficit at 72 h (Fig. 1d) were observed. Table 1 Kinetics of neurological dysfunction in IVIG-treated, C1-INH-treated and control mice following MCAO 0.05 compared to IVIG vehicle ** 0.05 compared to C1-INH vehicle # 0.05 compared to I/R only C1-INH Decreases Infarction Size and Neurological Deficit in Mice Subjected to Brain I/R Injury A single dose of C1-inhibitor (Berinert, CSL Behring) at 300 U/kg administered 30 min before ischemia and during reperfusion at 1 h, 3 SRI-011381 hydrochloride h and 6 h post-ischemia, significantly reduced brain infarction size relative to vehicle-treated mice by 69, 48, 50 and 67%, respectively (Fig. 2a). C1-INH treatment also significantly reduced practical impairment, as the neurological deficit scores (at 48.