title: Dynamic imaging of pancreatic nuclear factor κB (NF-κB) activation in live mice using adeno-associated virus (AAV) infusion and bioluminescence.
abstract: Nuclear factor κB (NF-κB) is an important signaling molecule that plays a critical role in the development of acute pancreatitis. Current methods for examining NF-κB activation involve infection of an adenoviral NF-κB-luciferase reporter into cell lines or electrophoretic mobility shift assay of lysate. The use of adeno-associated viruses (AAVs) has proven to be an effective method of transfecting whole organs in live animals. We examined whether intrapancreatic duct infusion of AAV containing an NF-κB-luciferase reporter (AAV-NF-κB-luciferase) can reliably measure pancreatic NF-κB activation. We confirmed the infectivity of the AAV-NF-κB-luciferase reporter in HEK293 cells using a traditional luciferase readout. Mice were infused with AAV-NF-κB-luciferase 5 weeks before induction of pancreatitis (caerulein, 50 μg/kg). Unlike transgenic mice that globally express NF-κB-luciferase, AAV-infused mice showed a 15-fold increase in pancreas-specific NF-κB bioluminescence following 12 h of caerulein compared with baseline luminescence (p < 0.05). The specificity of the NF-κB-luciferase signal to the pancreas was confirmed by isolating the pancreas and adjacent organs and observing a predominant bioluminescent signal in the pancreas compared with liver, spleen, and stomach. A complementary mouse model of post-ERCP-pancreatitis also induced pancreatic NF-κB signals. Taken together these data provide the first demonstration that NF-κB activation can be examined in a live, dynamic fashion during pancreatic inflammation. We believe this technique offers a valuable tool to study real-time activation of NF-κB in vivo.
authors: From the Department of Pediatrics and.
authors: Abrahim I
authors: AI
authors: Orabi
authors: From the Department of Pediatrics and.
authors: Swati
authors: S
authors: Sah
authors: From the Department of Pediatrics and.
authors: Tanveer A
authors: TA
authors: Javed
authors: From the Department of Pediatrics and.
authors: Kathryn L
authors: KL
authors: Lemon
authors: From the Department of Pediatrics and.
authors: Misty L
authors: ML
authors: Good
authors: Surgery, Children's Hospital of Pittsburgh of UPMC and the University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15224.
authors: Ping
authors: P
authors: Guo
authors: Surgery, Children's Hospital of Pittsburgh of UPMC and the University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15224.
authors: Xiangwei
authors: X
authors: Xiao
authors: Surgery, Children's Hospital of Pittsburgh of UPMC and the University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15224.
authors: Krishna
authors: K
authors: Prasadan
authors: Surgery, Children's Hospital of Pittsburgh of UPMC and the University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15224.
authors: George K
authors: GK
authors: Gittes
authors: From the Department of Pediatrics and.
authors: Shunqian
authors: S
authors: Jin
authors: From the Department of Pediatrics and sohail.husain@chp.edu.
authors: Sohail Z
authors: SZ
authors: Husain
mesh_terms: D000818:Animals; D002108:Ceruletide / Q000378:metabolism; D000229:Dependovirus / Q000235:genetics* / Q000502:physiology; D057809:HEK293 Cells; D006801:Humans; D008156:Luciferases / Q000235:genetics; D008163:Luminescent Measurements; D051379:Mice; D008822:Mice, Transgenic; D057054:Molecular Imaging; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D009928:Organ Specificity; D010179:Pancreas / Q000378:metabolism* / Q000821:virology*; D015398:Signal Transduction
chemical_list: D016328:NF-kappa B; D002108:Ceruletide; D008156:Luciferases
keywords: Bioluminescence; Mouse; NF-κB Transcription Factor; Pancreas; Transfection