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Rainbow trout (Oncorhynchus mykiss) interleukin-1 beta plasmid DNA generates inflammation in vivo and demonstrates adjuvant potential for vaccine against infectious hematopoietic necrosis virusJordan Hayes, Dawn Christine 02 March 2000 (has links)
Graduation date: 2000
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DNA vaccines encoding the glycoprotein genes of spring viremia of carp virus, snakehead rhabdovirus, or infectious hematopoietic necrosis virus induce protective immunity in rainbow trout (Oncorhynchus mykiss) against an infectious hematopoietic necrosis virus lethal challengeDrennan, John D. 08 July 1998 (has links)
Recent advances in DNA vaccine technology has brought about a promising
strategy for the control of viruses that contain surface membrane glycoproteins. This type
of vaccine involves the intramuscular injection of a bacterial plasmid containing a gene
encoding a viral protein. The strategy uses eukaryotic processing of the protein as would
naturally occur during a viral infection. In this study, plasmid DNA encoding the
glycoproteins of infectious hematopoietic necrosis virus (pcDNA3-IHNV-g), snakehead
rhabdovirus (pcDNA3-SHRV-g), or spring viremia of carp virus (pcDNA3-SVCV-g) was
injected into the skeletal muscle of rainbow trout fry. At 30 days post-vaccination, fish
were challenged with IHNV. Protection against IHNV was observed among all DNA
vaccinated groups. Fish injected with plasmid pcDNA3-IHNV-g, pcDNA3-SHRV-g, or
pcDNA3-SVCV-g had relative survival rates of 93.2%, 98.3% and 94.9%, respectively.
The mechanisms for the viral mediated resistance induced by these glycoprotein based DNA vaccines is unknown. A parallel study conducted by Dr. Carol Kim on the production of Mx proteins in these fish indicates that the observed protection might be a consequence of the stimulation of interferon. / Graduation date: 1999
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Rainbow trout cystatin C : gene expression, heterologous production and characterizationLi, Fugen 17 July 1998 (has links)
Rainbow trout cystatin C cDNA has been isolated from
trout liver. The full-length cystatin cDNA (674 bp)
included the 5'untranslated region and the polyadenylation
signal sequence AATAAA in the 3' region. Translation of
the cDNA defines 132 amino acid residues. Comparison of
the amino acid sequence with those of family 2 cystatins
indicates that the 21 amino acids at the N-terminal end is
a signal peptide necessary for cystatin secretion, and the
remaining 111 amino acids represent mature cystatin. Four
cysteine residues in the cystatin may form two disulfide
bonds producing a molecule with the properties of a family
2 cystatin.
Trout cystatin C gene expression was analyzed by
Northern blot. This gene is expressed at various levels in
all tissues examined. This difference may reflect
differences in degree of regulation of cysteine proteinase
activities. A high level of trout cystatin C expressed in
trout hepatic tissue or cell cultures suggested that
cystatin C expression might be related to tumorigenesis.
Southern blot of trout genomic DNA showed that the copy
number of the trout cystatin gene is probably one per
haploid genome.
Trout cystatin C was expressed in E. coli at a yield
of 3-5 mg/L culture, but no inhibitory activity was
detected for the untreated recombinant protein. However,
after refolding, recombinant cystatin C displayed
inhibitory activity against papain. The dissociation
constant of recombinant cystatin C against papain is 1.2 x
10������ nM, similar to that of human cystatin C. Trout
cystatin C was also expressed in yeast cells, but no
inhibitory activity was detected either. No cystatin C was
secreted in the yeast expression system using either the
trout cystatin C secretion signal, or the yeast invertase
secretion signal. The expression levels of trout cystatin
C in our expression systems are still low for industrial
requirements. Therefore, further investigation will be
needed to construct more efficient expression systems and
vectors for trout cystatin C heterologous production. / Graduation date: 1999
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Developmental of a novel affinity ELISA and ita application to the analysis of affinity maturation in troutShapiro, David A. 07 December 1995 (has links)
Graduation date: 1996
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Analysis of immune modulators in rainbow trout (Oncorhynchus mykiss)Mourich, Dan V. 20 March 1996 (has links)
The immune systems of various teleost fish have been studied in some detail for
the past several decades. One aspect of fish immunity, that of endogenously produced
modulating factors, has recently received a great deal of attention. Understanding the
functions and roles of endogenous factors that regulate fish immunity is paramount to
expanding the fields of fish immunology and vaccinology. It is know that several
lymphoid cell derived factors are detectable in in vitro cell culture systems and exhibit
immune modulating effects similar to well studied proteins in mammals. However in
comparison, few genes or gene products involved in the modulation of the trout immune
responses have been isolated, cloned and characterized.
The studies described herein were designed to isolate specific genes from rainbow
trout (0ncorhynchus mykiss) and characterize their involvement in the modulation and
regulation of the trout immune system. Two distinct genes were isolated cloned and
sequenced. The first, non-specific cytotoxic cell enhancement factor (NCEF) gene is
closely related to a human gene termed "natural killer enhancement factor" (NKEF) which
is important in the modulation of human natural killer cell activity. The second gene is
closely related to a group of recently characterized mammalian genes involved in the signal
transduction of cytokines termed "STATs". The role of these genes and their respective
protein products will be examined and discussed.
The antigenic structures of the fish proteins (NCEF and STAT5) were examined
by western blot and immunohistochemistry. Monoclonal antibodies derived against the
respective human proteins were found to cross react with the corresponding trout proteins,
demonstrating antigenic relatedness. The monoclonal regents were also used to analyze
the expression of these proteins in fish cells of lymphoid and non lymphoid origin.
In vitro cell culture analysis was used to determine the effects and roles of NCEF
and STAT5 gene products in the trout immune system. The cytolytic and apoptotic killing
activities of spleen, head kidney and peripheral blood leukocytes were found to be
enhanced by NCEF. Mitogenic stimulation of peripheral blood lymphoid cells resulted in
the trout STAT5 protein binding to a know sequences contained with in the promoters of
genes transcriptionally activated in response to cytokine exposure. / Graduation date: 1996
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Congener-specific disposition of polychlorinated biphenyls in rainbow troutFoster, Eugene P. 08 March 1996 (has links)
Graduation date: 1996
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Water quality modulation of aluminum toxicity to rainbow trout (Oncorhynchus mykiss) : biological and physiological approachesGundersen, Deke T. 13 December 1994 (has links)
Graduation date: 1995
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Modulation of respiratory burst activity of trout (Oncorhynchus mykiss) phagocytic cellsGalvan, Irja S. 29 April 1994 (has links)
Graduation date: 1995
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Chlorophyllin anticarcinogenesis in the rainbow trout modelBreinholt, Vibeke 21 April 1994 (has links)
Graduation date: 1994
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The genetic effect of stocking and population structure of brook trout (Salvelinus fontinalis) in the Beef River /Callen, Kevin. January 1983 (has links)
Thesis (M.S.)--University of Wisconsin -- La Crosse, 1983. / Includes bibliographical references (leaves 66-70).
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