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Title: Phenotype and cytokine expression profiles of ovine dendritic cells migrating to lymph nodes
Author: Bailey, Samantha L.
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 2005
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Dendritic cells function at the interphases of innate and adaptive immunity. The ability of dendritic cells (DC) to control the responses of effector T cells, B cells and NK cells are due in part to cell contacts and DC cytokines. DC are not an homogenous population, and different sub-sets may have different roles in vivo. Cannulation of the ‘pseudoafferent’ lymphatics is one of the few methods of obtaining differentiated DC ex vivo. This study describes the properties of two discrete ovine lymph DC populations defined by expression of signal regulatory protein-α (SIRPα). SIRPα+ DC were essentially CD58+, CD11a+, WC6+, CD54RA-, expressed CD16, CD14, CD206, CD4 and CD8 at low levels or no minor sub-populations, and as a single population expressed higher levels of CD11c, CD40, CD80, CD86 and MHC II than did SIRPα- DC. SIRPα- DC were CD16-, CD14-, CD206-, CD4-, CD8-, CD11a+ and displayed heterogeneity of expression of CD58, CD11c, WC6 and CD45RA. Importantly, freshly isolated SIRPα+ DC expressed IL-12 p40 and IL-18 mRNA, whereas SIRPα-were negative for these transcripts. Neither IL-6 or IL10 were detected in the DC populations by PCR. It appears therefore that the SIRPα DC populations may instigate different lymphocyte responses in vivo. To support studies in ovine model systems, sheep interleukin-18 (IL-18) was cloned and sequenced. By inducing the production of interferon gamma, IL-18 is an important regulator of T cell immunity. IL-18 was utilised with ten other sheep cytokines for the development of a multiprobe RNase protection assay (RPA). Standardisation analysis showed that the RPA could reliably quantify levels of IL-1β, IL-4, IL-6, IL-10, IL-12 p40, IL-18, GM-CSF, IFNγ and TGFβ. IL-8 and TNFα-specific riboprobes require optimisation for use with the developed protocol. The RPA will complement studies of ovine disease immunopathology.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available