Previously we determined that BHV-5 gE is important for BHV-5 neurovirulence. Envelope glycoproteins E (gE) and gI of alphaherpesviruses are important for the pathogenesis in vivo. Progress 10/01/99 to 09/30/04 Outputs Bovine herpesvirus type 5 (BHV-5) is an alphaherpesvirus that causes fatal encephalitis in calves. Neurovirulence and neuroinvasive properties of these viruses will be analyzed by intranasal and olfactory bulb inoculations. To dissect the gD and gE coding regions with respect to neurovirulence and neuroinvasiveness function, we will construct chimeric BHV-5 viruses in which only portions of gE or gD will be expressed. Following the injection we will carefully analyze the viral spread and We also plan to inject BHV-1, BHV-5 and their recombinants into the olfactory bulb by stereotaxic injections. Following infection, the rabbits will be euthanized at different time points and the brains will be processed for immunohistochemistry and the skulls will be processed for double immunofluorescence labeling and electron microscopy. To address the role of gD, we will use gD-deleted and gD-exchanged BHV-5 (contain BHV-1 gD), and gD-exchanged BHV-1 (contain BHV-5 gD) viruses to infect the rabbits. between 1-5 days, infected and uninfected (control) rabbits will be euthanized and their skull segment extending from mid-maxillary region to just proximal to orbits will be processed for histopathology, double immunofluorescence labeling (for virus and neuronal marker) and electron microscopy. At different times after infections, i.e.
Project Methods We will infect rabbits intranasally with either BHV-1 or BHV-5.
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