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Title: The role of interleukin-8 in inflammatory responses of fish : studies in rainbow trout (Oncorhynchus mykiss)
Author: Harun, Nor Omaima
ISNI:       0000 0001 3542 1284
Awarding Body: University of Aberdeen
Current Institution: University of Aberdeen
Date of Award: 2006
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The predicated rainbow trout Oncorhynchus mykiss mature intereleukin-8 peptide has been produced as a recombinant protein in E. coli; interleukin-8 (rIL-8a) and a mutated form lacking a receptor binding motif (rIL-8b) were studied initially in vitro and in vivo. The bioactivity of this molecule has been studied using trout head kidney cell preparations, using a microchemotaxis chamber designed to study the migration of cell populations in vitro. Various concentration gradients were tested (0.1, 1.0 and 10.0ng/ml) to determine optimal assay conditions and suitable attractants for such an assay and the numbers of cells migrating counted. The recombinant trout IL-8 and the mutated recombinant IL-8 induced specific dose-dependent in vitro chemotaxis of leukocytes at doses as low as 1.0ng/ml. To study respiratory burst kinetics; optimal final concentration of PMA and Ferricytochrome C were established at 1μg ml-1 and 2mg ml-1 for stimulation of the respiratory burst and detection of superoxide onion respectively. In vitro dose-response studies demonstrated an increased stimulation with an optimal concentration of the recombinant protein of 1.0ng ml-1. Following on from these preliminary studies, the recombinant proteins were injected into fish, and the numbers of leukocyte subpopulations elicited in exudates and blood, and their activation state determined. The aim of this experiment was to investigate the ability of trout IL-8 to modulate the expression of certain immune parameters and disease resistance in vivo. Immunostimulatory activity in vivo was elucidated with respect to induction of a localised immune response as a consequence of intraperitoneal peptide administration, where the responses characterised included leukocyte migration to the peritoneal cavity.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available