Induction and characterization of endotoxin tolerance in equine peripheral blood mononuclear cells in vitro

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dc.contributor.authorFrellstedt, Lindaen
dc.contributor.committeechairFurr, Martin O.en
dc.contributor.committeememberMcKenzie, Harold C. IIIen
dc.contributor.committeememberBarrett, Jennifer G.en
dc.contributor.departmentBiomedical and Veterinary Sciencesen
dc.date.accessioned2014-03-14T21:43:05Zen
dc.date.adate2010-09-10en
dc.date.available2014-03-14T21:43:05Zen
dc.date.issued2010-06-23en
dc.date.rdate2010-09-10en
dc.date.sdate2010-08-19en
dc.description.abstractEndotoxemia 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.degreeMaster of Scienceen
dc.identifier.otheretd-08192010-172631en
dc.identifier.sourceurlhttp://scholar.lib.vt.edu/theses/available/etd-08192010-172631/en
dc.identifier.urihttp://hdl.handle.net/10919/44307en
dc.publisherVirginia Techen
dc.relation.haspartFrellstedt_L_T_2010.pdfen
dc.rightsIn Copyrighten
dc.rights.urihttp://rightsstatements.org/vocab/InC/1.0/en
dc.subjectEndotoxinen
dc.subjectendotoxin toleranceen
dc.subjectinterleukin-12en
dc.subjectinterleukin-10en
dc.subjectequineen
dc.titleInduction and characterization of endotoxin tolerance in equine peripheral blood mononuclear cells in vitroen
dc.typeThesisen
thesis.degree.disciplineBiomedical and Veterinary Sciencesen
thesis.degree.grantorVirginia Polytechnic Institute and State Universityen
thesis.degree.levelmastersen
thesis.degree.nameMaster of Scienceen

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