Use this URL to cite or link to this record in EThOS: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.650935
Title: Studies on ovine interleukin-1
Author: Fiskerstrand, C. E.
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 1994
Availability of Full Text:
Full text unavailable from EThOS.
Please contact the current institution’s library for further details.
Abstract:
Interleukin-1 (IL-1) is a key mediator of infection, inflammation and immunity and two different IL-1 proteins are known to exist, IL-1α and IL-1β. Each is synthesised as a proprotein, Mr = 31kD, which is subsequently cleaved to yield the mature protein, Mr=17.5kD. Whereas both forms of IL-1α show equivalent biological activities, IL-1β requires cleavage with resultant conformational change for optimal activity. IL-1 has been extensively studied in human and murine systems but at the time this project was initiated, nothing was known about its actions in the sheep and no reagents were available with which to study ovine IL-1. This thesis describes the successful cloning and expression of biologically active ovine IL-1α and IL-1β and their use in determining IL-1 receptor (IL-1R) expression by ovine alveolar macrophages (Mφ) and afferent lymph dendritic cells (DC). Lipopolysaccharide (LPS) stimulated Mφ were used as the source of IL-1 mRNA. The specific IL-1 cDNAs were amplified by polymerase chain reaction, cloned into pTZ18R/19R vectors and sequenced. Ovine IL-1α and IL-1β were found to be 97% and 96% identical to their bovine counterparts and 81% and 78% identical, respectively, to the human sequences. Translation of the nucleotide sequences shows that ovine IL-1α has 97% and 72% identity with bovine and human IL-1α respectively and ovine IL-1β has 95% and 62% identity with bovine and human IL-1β respectively. Use of the cloned IL-1cDNAs for northern blot analysis of IL-1 mRNA production by Mφ, showed IL-1α mRNA to reach maximal levels at around 6h and IL-1β mRNA at around 4h after LPS stimulation. The proprotein (p) and mature protein (m) forms of ovine IL-1α and IL-1β have been expressed as fusion proteins with yeast p1, using the Ty-vlp system of British Biotechnology Ltd.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.650935  DOI: Not available
Share: