The milk protein promoter is a useful tool for developing a rat with tolerance to a human protein.
Biopharmaceuticals intended for humans are immunogenic in animals. Antibodies associated with their administration make it difficult to perform repeated-dose pharmacology and toxicology studies in animals. Despite suggestions to solve this problem with transgenic animal technology, an effective strategy has not yet been reported. The objective of the present study was to provide an efficient strategy to develop rats tolerant to biopharmaceuticals such as human gene-based proteins. The present study used transgenic rat lines (lines 311-6, 308-5, and 305-1) carrying a fusion gene designed to express the human growth hormone (hGH) gene under the control of the bovine alphaS1 casein gene promoter. Three lactating females with the transgene, produced approximately 4 mg/ml, 300 microg/ml, and 10 ng/ml in their milk. Male 8-week-old rats from these three lines were immunized with hGH three times (week 0, 1, and 3 ) and the production of antibodies against hGH in their sera were examined at week 4. While the hGH serum antibody titers increased over 1000-fold in wild-type control rats, there was no detectable antibody against hGH in the sera of these three transgenic lines. Human growth hormone in their sera was undetectable (lines 308-5 and 305-1) or much lower than the endogenous biologic level of rat growth hormone (line 311-6). Importantly, lines 308-5 and 305-1 developed tolerance to hGH without detectable hGH in their sera and these lines will be very useful for the repeated dose pharmacology and toxicology studies. These results suggest that a milk protein promoter can be a useful tool to develop transgenic rats that are tolerant to biopharmaceuticals intended for humans.
Takahashi
R
R
YS New Technology Institute Inc, Tochigi, Japan. LEH03041@nifty.ne.jp
Ueda
M
M
D000818:Animals; D030801:Animals, Genetically Modified; D001688:Biological Products / Q000097:blood / Q000493:pharmacokinetics*; D002364:Caseins / Q000235:genetics; D002417:Cattle; D018014:Gene Transfer Techniques; D019382:Human Growth Hormone / Q000097:blood / Q000493:pharmacokinetics*; D006801:Humans; D008297:Male; D008894:Milk Proteins / Q000235:genetics*; D011401:Promoter Regions, Genetic; D051381:Rats; D013997:Time Factors
D001688:Biological Products; D002364:Caseins; D008894:Milk Proteins; D019382:Human Growth Hormone