MAF Fellowship Training Grant - use of recombinant proteins to identify antibody responses associated with Equine Proliferative Enteropathy.

Hypothesis/Objectives: Our hypothesis is that the antibody response to specific L. intracellularis antigens can be used to identify EPE affected horses. This will be accomplished by creating a DNA expression library (Objective #1), which we will use to obtain L. intracellularis recombinant proteins for immunoblot screening (Objective #3). We will also use 2D-SDS-PAGE gel electrophoresis of whole L. intracellularis (Objective #2) blotted with serum from L. intracellularis-exposed horses to identify immunoreactive proteins (Objective #3). The immunoreactive recombinant proteins will then be used to screen sera from horses representing clinically EPE affected, exposed but not clinically affected, and non-exposed horses (Objective #4). This will allow us to identify antibody responses to specific proteins that may be diagnostic for either clinical EPE or protective immunity. Proteins that do demonstrate variable immunoreactivity will be explored for possible use in a new ELISA.