Induction and characterization of endotoxin tolerance in equine peripheral blood mononuclear cells in vitro
dc.contributor.author | Frellstedt, Linda | en |
dc.contributor.committeechair | Furr, Martin O. | en |
dc.contributor.committeemember | McKenzie, Harold C. III | en |
dc.contributor.committeemember | Barrett, Jennifer G. | en |
dc.contributor.department | Biomedical and Veterinary Sciences | en |
dc.date.accessioned | 2014-03-14T21:43:05Z | en |
dc.date.adate | 2010-09-10 | en |
dc.date.available | 2014-03-14T21:43:05Z | en |
dc.date.issued | 2010-06-23 | en |
dc.date.rdate | 2010-09-10 | en |
dc.date.sdate | 2010-08-19 | en |
dc.description.abstract | Endotoxemia is responsible for severe illness in horses. Individuals can become unresponsive to the endotoxin molecule after an initial exposure; this phenomenon has been called developing a state of "endotoxin tolerance" (ET). ET has been induced in horses in vivo; however, cytokine expression associated with ET has not been investigated. The purpose of this study was to develop and validate a method for inducing ET in equine peripheral blood mononuclear cells (PBMCs) in vitro, and to describe the cytokine profile which is associated with the ET. Blood was collected from 6 healthy horses and PBMCs were isolated. ET was induced by culturing cells with three concentrations of endotoxin given to induce ET, and evaluated after a second dose of endotoxin given to challenge the cells. The relative mRNA expression of IL-10 and IL-12 was measured by use of quantitative PCR. ET was induced in all cells (n=6) exposed to the 2-step endotoxin challenge. In PBMCs treated with 1.0 ng/ml of endotoxin followed by challenge with 10 ng/ml of endotoxin, the relative mRNA expression of IL-10 in tolerized cells was not different from positive control cells. In contrast, the relative mRNA expression of IL-12 in tolerized cells was decreased by 15-fold after the second endotoxin challenge compared with positive control cells. This experiment demonstrated a reliable method for the ex vivo induction of ET in equine PBMCs. A marked suppression of IL-12 production is associated with ET. The production of IL-10 was not altered in ET in our model. | en |
dc.description.degree | Master of Science | en |
dc.identifier.other | etd-08192010-172631 | en |
dc.identifier.sourceurl | http://scholar.lib.vt.edu/theses/available/etd-08192010-172631/ | en |
dc.identifier.uri | http://hdl.handle.net/10919/44307 | en |
dc.publisher | Virginia Tech | en |
dc.relation.haspart | Frellstedt_L_T_2010.pdf | en |
dc.rights | In Copyright | en |
dc.rights.uri | http://rightsstatements.org/vocab/InC/1.0/ | en |
dc.subject | Endotoxin | en |
dc.subject | endotoxin tolerance | en |
dc.subject | interleukin-12 | en |
dc.subject | interleukin-10 | en |
dc.subject | equine | en |
dc.title | Induction and characterization of endotoxin tolerance in equine peripheral blood mononuclear cells in vitro | en |
dc.type | Thesis | en |
thesis.degree.discipline | Biomedical and Veterinary Sciences | en |
thesis.degree.grantor | Virginia Polytechnic Institute and State University | en |
thesis.degree.level | masters | en |
thesis.degree.name | Master of Science | en |
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