Multiorgan Nuclear Factor Kappa B Activation in a Transgenic Mouse Model of Systemic Inflammation

Abstract

We utilized a line of transgenic mice expressing Photinus luciferase complementary DNA (cDNA) under the control of a nuclear factor kappa B (NF-kappaB)-dependent promoter (from the 5' human immunodeficiency virus-1 [HIV-1] long terminal repeat) to examine the role of NF-kappaB activation in the pathogenesis of systemic inflammation induced by bacterial endotoxin (lipopolysaccharide [LPS]). After intraperitoneal injection of E. coli LPS, these mice displayed a time- and dose-dependent, organ-specific pattern of luciferase expression, showing that NF-kappaB-dependent gene transcription is transiently activated in multiple organs by systemic LPS administration. Luciferase expression in liver could be specifically blocked by intravenous administration of replication-deficient adenoviral vectors expressing a dominant inhibitor of NF-kappaB (IkappaB-alphaDN), confirming that luciferase gene expression is a surrogate marker for NF-kappaB activation in this line of mice. After treatment with intraperitoneal LPS, the mice were found to have increased lung tissue messenger RNA (mRNA) expression of a variety of cytokines that are thought to be NF-kappaB-dependent, as well as elevated serum concentrations of presumed NF-kappaB-dependent cytokines. In lung tissue homogenates, a close correlation was identified between luciferase activity and KC levels. These studies show that systemic treatment with LPS orchestrates a multiorgan NF-kappaB-dependent response that likely regulates the pathobiology of systemic inflammation.

Prostaglandin I2 Analogs Inhibit Proinflammatory Cytokine Production and T Cell Stimulatory Function of Dendritic Cells

“…NF-B is a critical transcription factor in the molecular regulation of inflammatory cytokine and chemokine expression (27). Known NF-B-dependent genes include IL-12, TNF-␣, IL-6, MCP-1, KC, and IL-1␤ (12,27,41,42). Elevation of intracellular cAMP by cAMP-elevating agents decreased LPS-induced NF-B function as shown in this study and in several other studies (37,(43)(44)(45) and this effect was partially attenuated by Rp-8-Br-cAMPS, suggesting that PGI 2 analogs may suppress NF-B function by activating the cAMP-signaling pathway.…”

“…Age-and sex-matched C57BL/6J mice (from The Jackson Laboratory) were used as wild-type control mice (IP ϩ/ϩ ). NF-B reporter transgenic mice, referred to as HLL (HIV-long terminal repeat (LTR)/ luciferase) mice on a C57BL/6/DBA background, were generated as described (12). HLL mice carry the Photinus luciferase gene cDNA driven by the proximal 5Ј HIV (HIV-1) LTR promoter (12,13).…”

“…NF-B reporter transgenic mice, referred to as HLL (HIV-long terminal repeat (LTR)/ luciferase) mice on a C57BL/6/DBA background, were generated as described (12). HLL mice carry the Photinus luciferase gene cDNA driven by the proximal 5Ј HIV (HIV-1) LTR promoter (12,13). The proximal HIV-LTR contains two NF-B motifs and is a well-characterized NF-B-responsive promoter (14).…”

“…To test the hypothesis that the anti-inflammatory effect of PGI 2 was associated with inhibition of NF-B activation, we examined the effect of PGI 2 analogs on NF-B transcriptional activity in BMDCs that were derived from NF-B reporter HLL mice. In cells from these mice, luciferase expression can be assayed as a measure of NF-B activity (12,13).…”

Airway Epithelium Controls Lung Inflammation and Injury through the NF-κB Pathway

“…I B-␣DN-transactivated (DNTA) transgenic mice. To tag the I B-␣ dominant inhibitor (I B-␣DN) (8,11,12), a 1.35-kb BamHI/DraI fragment was excised from pCMX-pp40 and blunt-end ligated into BamHI/EcoRVdigested pEF4/Myc-HisA (Invitrogen Life Technologies). The I B-␣DN-Myc-His-tagged fragment was then excised by BamHI digest, fill-in of the overhanging ends and PmeI digestion.…”