Development of immunological memory in rainbow trout (Oncorhynchus mykiss) and aflatoxin B₁ modulation of the response Public Deposited

http://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/bn999b58v

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  • Immunological memory has been shown to occur unequivocally in mammals. However, controversy exists regarding the capability of fish to elicite a memory response. This study demonstrates that memory does occur rainbow trout, Oncorhynchus rnykiss. Trout injected with a T-dependent antigen, trinitrophenylated keyhole-limpet hemocyanin, (TNP-KLH) (100 μg/fish) were able to produce a primary serum antibody response to TNP. This was demonstrated by a quantitative ELISA specifically developed for this purpose. After a second injection of antigen (20 μg/fish), an increase in antibody titer developed to TNP along with a shorter lag period, as is characteristic of a mammalian secondary response. The antibodies produced to TNP were further analyzed by two different ELISAs, avidity and inhibition ELISAs. Avidity measurements, which reflects both the effects of affinity and valency on the strength of antibody binding, demonstrated no change in avidity during the primary or secondary serum antibody response. The inhibition ELISA it was possible to demonstrate subtle changes in the fine specificity of the antibody populations by using a sufficiently diverse battery of TNP-lysine analogues. In vivo priming of trout with TNP-KLH produced cells which were capable of producing a heightened in vitro plaque-forming cell response to TNP-LPS, a T-independent antigen, and TNP-KLH a T-dependent antigen. This is similar to what has been reported in mammals and demonstrates that fish are capable of expressing memory to T-independent and T-dependent forms of antigen. The modulation of the memory response was also monitored after a short embryonic exposure of aflatoxin B i in order to determine if this environmental toxicant could affect the primary and/or memory antibody response to TNP. Neither in vivo nor in vitro primary responses were affected by the mycotoxin, but the ability to produce a memory response was eliminated. A change in avidity due to aflatoxin exposure was not witnessed, but the fine specificities of the antibodies were altered as was evidenced by examination of the relative affinities for the analogues of TNP.
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