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Optimized Interferon-gamma ELISpot Assay to Measure T Cell Responses in the Guinea Pig Model after Vaccination
Published on: January 20, 2019
Genetic immunization with codon-optimized equine infectious anemia virus (EIAV) surface unit (SU) envelope protein
R Frank Cook1, Sheila J Cook, Pamela S Bolin
1Department of Veterinary Science, Gluck Equine Research Center, University of Kentucky, Lexington, KY 40546, USA. rfcook1@uky.edu
Abstract:
In the context of DNA vaccines the native equine infectious anemia virus (EIAV)-envelope gene has proven to be an extremely weak immunogen in horses probably because the RNA transcripts are poorly expressed owing to an unusual codon-usage bias, the possession of multiple RNA splice sites and potential adenosine-rich RNA instability elements. To overcome these problems a synthetic version of sequences encoding the EIAV surface unit (SU) envelope glycoprotein was produced (SYNSU) in which the codon-usage bias was modified to conform to that of highly expressed horse and human genes. In transfected COS-1 cell cultures, the steady state expression levels of SYNSU were at least 30-fold greater than equivalent native SU sequences. More importantly, EIAV-specific humoral and lymphocyte proliferation responses were induced in ponies immunized with a mammalian expression vector encoding SYNSU. However, these immunological responses were unable to confer protection against infection with a virulent EIAV strain.
