A narrow time-window for access to the brain by exogenous protein after immunological targeting of a blood-brain barrier antigen

Abstract

The endothelial barrier antigen (EBA) is a membrane protein expressed by endothelial cells of the rat blood–brain barrier (BBB). A previous short-term non-recovery study demonstrated that immunological targeting of EBA by intravenous administration of a monoclonal antibody (anti-EBA) led to acute opening of the BBB to exogenous and endogenous tracers. The aims of the present study were to determine whether opening of the BBB was reversible and compatible with survival, and whether a “therapeutic window” existed. A single intravenous injection of one of three doses (high, medium and low) of anti-EBA was used. Animals were allowed to survive for periods ranging from 17 min to 4 days. The tracer horseradish peroxidase (HRP) was administered intravenously 10 min before perfusion fixation, and its distribution was assessed in Vibratome sections of the brain and spinal cord. Leakage of HRP into the central nervous system was dose- and time-dependent. The medium dose produced incipient HRP leakage at 17 min and widespread pronounced leakage at 30 min. Progressive reduction in HRP permeability occurred from 45 min to 2 h, with barrier restoration by 3 h. At all subsequent time intervals (6 h–4 days) the BBB remained impermeable to HRP. The low and high doses produced less and greater HRP leakage, respectively, but restoration of the barrier still occurred at 3 h. The high dose, however, produced a number of deaths. Animals treated with an isotype control antibody showed no HRP leakage at comparable time intervals. The results indicated that (1) this model was compatible with survival, (2) opening of the BBB was monophasic and transient, occurring during a narrow “time-window”, and (3) the barrier, once reconstituted, maintained its integrity.