Acute phase proteins in rainbow trout (Oncorhynchus mykiss) Public Deposited

http://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/s4655j643

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  • The acute phase response (APR) includes an early response of the innate immune system to an inflammatory agent. Acute phase proteins (APP) are produced at increased rates in the liver during this response. The APR has been well characterized in mammals, but it is unclear to what degree teleosts mount a similar response. This thesis describes research conducted to characterize this response in rainbow trout. We further developed and used a Tandem Crossed Immunoelectrophoresis Technique (TCIEP). With this we were able to determine that concentrations of at least three trout plasma proteins were higher several days after trout had been injected with Freund's Complete Adjuvant. One of these proteins was isolated and partially characterized, using electrospray mass spectrometry (ESI-MS), and tentatively identified as transferrin. Due to the limited amount of any protein that could be obtained from T-CIEPs, we switched to the use of 2D gel electrophoresis based on isoelectric focusing and SDS PAGE. When comparing such 2D gels containing trout plasma from before and after treatment, at least 5 of approximately 200 protein spots were increased in size. An N-terminal sequence was obtained from three of the increased spots, and degenerate primers constructed from one of those sequences; 5' and 3' RACE PCR revealed a 188 amino acid long polypeptide. It was 29% identical at the amino acid level with precerebellin from rat. The function of precerebellin is unknown but it appears likely that it is part of the innate immune system since it is unregulated in trout injected with Vibrio bacterin. This thesis has begun the characterization of the acute phase proteins in rainbow trout, but much work remains to be conducted.
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