Inducible and tissue-specific expression of rat alpha-1-acid glycoprotein in transgenic mice.

Abstract

alpha-1-Acid glycoprotein (AGP), which is produced in the mammalian liver and secreted into the blood-stream, is regulated by steroid hormones and by mediators of the acute phase response. In vitro transfection studies have shown that the response to glucocorticoids requires a cis-acting regulatory element, termed the glucocorticoid response element, that is located within 120 bp of the transcriptional start-site of the gene; induction by the acute phase mediators requires a different element, termed the distal regulatory element (DRE), that is located about 5 kb upstream of the start-site. To determine if these elements function in vivo, we have produced and characterized transgenic mice containing rat AGP gene constructs with and without the DRE. Five transgenic lines were produced from a 9.5-kb genomic AGP containing 4.7 kb of the 5' flanking region; this construct lacks the DRE. Another transgenic line was derived from a 10.7-kb clone that contains 5.3 kb of 5' flanking sequences including the DRE. All transgenic mice produced high levels of immunologically detectable rat AGP in the circulation, comparable to or in excess of that found in normal rats. There were correspondingly high concentrations of rat AGP transcripts in the liver. Transgene expression in all lines was induced in response to dexamethasone and during acute inflammation resulting from LPS treatment. The DRE-containing transgene underwent a greater induction in response to LPS than to dexamethasone; the transgene lacking the DRE responded similarly to both treatments. In cultured primary hepatocytes, the DRE-containing transgene was induced by the acute phase cytokines IL-1 and IL-6, and by dexamethasone, administered individually or in combination; the transgene lacking the DRE responded only to dexamethasone, and was not affected by the peptide hormones. Together, these results provide in vivo evidence supporting the notion that a minimum of two upstream sequences are responsible for the inflammatory induction of rat AGP. One element, which is located within the smaller 9.7-kb restriction fragment, is responsive to glucocorticoids and is likely to be the glucocorticoid response element located close to the transcriptional start site. The other element, the DRE, is located much further upstream and is responsible for imparting responsiveness to the acute phase cytokines.