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Matthew M. Quesnell1 , Yuzhi Zhang1 , David M. de Avila1 , Kevin P. Bertrand2 , Jerry J. Reeves1 *
Department of Animal Science, Washington State University, Pullman, WA 1
Department of Biophysics and Biochemistry, Washington State University, Pullman, WA 2
Genes for the hybrid ovalbumin-LHRH-7 and thioredoxin-LHRH-7 fusion proteins (each containing 7 LHRH inserts distributed at multiple sites) were constructed by cassette mutagenesis and oligonucleotide mismatch mutagenesis and expressed in E. coli as insoluble proteins. Constructs included a gene encoding for six histidine molecules on the C terminus of the proteins, allowing for nickel column affinity purification. Injection of mice was performed three times at four-week intervals with 10 µg of protein suspended in Z-max adjuvant. Ovalbumin-LHRH-7 and thioredoxin-LHRH-7 treated mice had significantly elevated concentrations of LHRH antibodies (20 ± 9 %, 49 ± 11 % vs .2 ± .2 %) respectively, compared to controls. These treated mice also had significantly lower testicular wt (144 ± 12 mg, 138 ± 9 mg vs 192 ± 5 mg), epididymal wt (38 ± 11 mg, 59 ± 5 mg vs 79 ± 3 mg) and vesicular gland/anterior prostate wt (153 ± 35 mg, 132 ± 36 mg, vs 258 ± 6 mg) compared to all controls. The body wt of mice treated with thioredoxin-LHRH-7 was significantly lower than in the control animals (25.5 ± .7 g vs 28.5 ± .4 g). Only 40% of the ovalbumin-LHRH-7 and 60% of the thioredoxin-LHRH-7 treated animals showed a biological response to the immunization. Examination of LHRH antibody affinities indicated large variation between animals. This could explain the apparent discrepancy sometimes seen between antibody binding concentrations and biological response. Because of the dramatic ability to induce anterior prostate hypoplasia, future studies are warranted to explore the potential use of these proteins as a treatment for carcinoma of the prostate.
This abstract is being presented on Monday, August 2 at 8:00 AM to 10:15 AM at CUB 2nd Floor Ballroom.