D E Frank1, F A Ahrens, T T Kramer. 1. Department of Microbiology, Immunology, and Preventive Medicine, College of Veterinary Medicine, Iowa State University, Ames 50011, USA.
Abstract
OBJECTIVE: To develop a model to study the kinetics and relative amounts of cytokines produced by liver cells during enteric infection. DESIGN: Salmonella enteriditis lipopolysaccharide (LPS)- or live S choleraesuis-stimulated isolated livers from clinically normal pigs and pigs with active acute phase response. ANIMALS: 7- to 14-day-old salmonellosis-free pigs, 4 to 12/group. PROCEDURE: Livers were removed and perfused with oxygenated Krebs-Henseleit solution for 30 minutes and with S choleraesuis or LPS added for 7 minutes. Livers were then perfused with 500 ml of fresh solution in a closed loop procedure for 180 minutes. Perfusate samples were collected for tumor necrosis factor-alpha (TNF alpha) and interleukin 6 (IL-6) bioassays. RESULTS: Tumor necrosis factor-alpha values remained constant during perfusion of normal livers and increased in those exposed to LPS. Interleukin 6 values increased in perfusate from normal livers from 30 to 150 minutes, then decreased. In livers from pigs with an active acute phase response, TNF alpha values were reduced; IL-6 appeared by 2 minutes and decreased after 25 minutes. CONCLUSIONS: Isolated livers could be kept viable for 3 hours, and IL-6 and TNF alpha could be measured by the bioassays used. CLINICAL RELEVANCE: Model can be used for studying and modifying the response of liver cells to infective agents.
OBJECTIVE: To develop a model to study the kinetics and relative amounts of cytokines produced by liver cells during enteric infection. DESIGN: Salmonella enteriditis lipopolysaccharide (LPS)- or live S choleraesuis-stimulated isolated livers from clinically normal pigs and pigs with active acute phase response. ANIMALS: 7- to 14-day-old salmonellosis-freepigs, 4 to 12/group. PROCEDURE: Livers were removed and perfused with oxygenated Krebs-Henseleit solution for 30 minutes and with S choleraesuis or LPS added for 7 minutes. Livers were then perfused with 500 ml of fresh solution in a closed loop procedure for 180 minutes. Perfusate samples were collected for tumor necrosis factor-alpha (TNF alpha) and interleukin 6 (IL-6) bioassays. RESULTS:Tumor necrosis factor-alpha values remained constant during perfusion of normal livers and increased in those exposed to LPS. Interleukin 6 values increased in perfusate from normal livers from 30 to 150 minutes, then decreased. In livers from pigs with an active acute phase response, TNF alpha values were reduced; IL-6 appeared by 2 minutes and decreased after 25 minutes. CONCLUSIONS: Isolated livers could be kept viable for 3 hours, and IL-6 and TNF alpha could be measured by the bioassays used. CLINICAL RELEVANCE: Model can be used for studying and modifying the response of liver cells to infective agents.
Authors: Justin E Juskewitch; Jeffrey L Platt; Bruce E Knudsen; Keith L Knutson; Gregory J Brunn; Joseph P Grande Journal: Sci Rep Date: 2012-12-04 Impact factor: 4.379