Figure 5.

RBC-targeted scFv-TM protects against cerebral ischemia-reperfusion injury and resulting neurologic deficits. Twenty minutes before carotid artery ligation, mice were treated with PBS (black bars), 18 nmol/kg sTM (1 mg/kg equivalent, white bars), or 18 nmol/kg scFv/TM (hashed bars) given via injection into the right jugular vein. The external carotid artery was then isolated and incised, a silicon-covered nylon filament was advanced via the proximal external carotid artery into the internal carotid artery, and the MCA was occluded for 30 min with subsequent filament withdrawal and reperfusion for 48 h using laser doppler to monitor cortical blood flow. The brain was cut into 2-mm-thick coronal blocks, and the sections were stained with 2,3,5-triphenyltetrazolium chloride to calculate the infarct volume. Residual neurologic function was assessed using a 4-point scale (see Materials and Methods). A, B) The results demonstrate that scFv/TM, but not sTM, significantly reduced infarct volume (A) and neurologic deficit (B) 48 h after MCA occlusion. C) Representative images demonstrating reduced area of infarct (pale coloration) (n = 3–5; means ± sem). *P < 0.05 vs. PBS control (1-tailed Student’s t test).