Immunoresponse of Coho salmon immunized with a gene expression library from Piscirickettsia salmonis

Alvaro Miquel, Ilse Müller, Pablo Ferrer, Pablo D T Valenzuela, Luis O. Burzio

Resultado de la investigación: Article

15 Citas (Scopus)

Resumen

We have used the expression library immunization technology to study the protection of Coho salmon Oncorhynchus kisutch to the infection with Piscirickettsia salmonis. Purified DNA from this bacterium was sonicated and the fragments were cloned in the expression vector pCMV-Bios. Two libraries were obtained containing 22,000 and 28,000 colonies and corresponding to approximately 8 and 10 times the genome of the pathogen, respectively. On average, the size of the inserts ranged between 300 and 1,000 bp. The plasmid DNA isolated from one of these libraries was purified and 20 μg were injected intramuscularly into 60 fish followed by a second dose of 10 μg applied 40 days later. As control, fish were injected with the same amount of DNA of the vector pCMV-Bios without insert. The titer of IgM anti-P. salmonis of vaccinated fish, evaluated 60 days post-injection, was significantly higher than that of the control group injected with the vector alone. Moreover, this response was specific against P. salmonis antigens, since no cross reaction was detected with Renibacterium salmoninarum and Yersinia ruckeri. The vaccinated and control fish were challenged 60 days after the second dose of DNA with 2.5 × 107 P. salmonis corresponding to 7.5 times the LD50. At 30 days post-challenge, 100% mortality was obtained with the control fish while 20% of the vaccinated animals survived. All surviving fish exhibited a lower bacterial load in the kidney than control fish. The expression library was also tested in Balb/c mice and it was found that the humoral immune response was specific to P. salmonis and it was dependent on the amount of DNA injected.

Idioma originalEnglish
Páginas (desde-hasta)313-323
Número de páginas11
PublicaciónBiological Research
Volumen36
N.º3-4
DOI
EstadoPublished - 1 ene 2003

Huella dactilar

Piscirickettsia
Piscirickettsia salmonis
Oncorhynchus kisutch
Gene Library
Gene expression
Fish
Fishes
Gene Expression
gene expression
fish
DNA
Libraries
Yersinia ruckeri
Renibacterium salmoninarum
Immunization
Bacterial Load
Lethal Dose 50
Cross Reactions
Pathogens
Humoral Immunity

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Citar esto

Miquel, Alvaro ; Müller, Ilse ; Ferrer, Pablo ; Valenzuela, Pablo D T ; Burzio, Luis O. / Immunoresponse of Coho salmon immunized with a gene expression library from Piscirickettsia salmonis. En: Biological Research. 2003 ; Vol. 36, N.º 3-4. pp. 313-323.
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abstract = "We have used the expression library immunization technology to study the protection of Coho salmon Oncorhynchus kisutch to the infection with Piscirickettsia salmonis. Purified DNA from this bacterium was sonicated and the fragments were cloned in the expression vector pCMV-Bios. Two libraries were obtained containing 22,000 and 28,000 colonies and corresponding to approximately 8 and 10 times the genome of the pathogen, respectively. On average, the size of the inserts ranged between 300 and 1,000 bp. The plasmid DNA isolated from one of these libraries was purified and 20 μg were injected intramuscularly into 60 fish followed by a second dose of 10 μg applied 40 days later. As control, fish were injected with the same amount of DNA of the vector pCMV-Bios without insert. The titer of IgM anti-P. salmonis of vaccinated fish, evaluated 60 days post-injection, was significantly higher than that of the control group injected with the vector alone. Moreover, this response was specific against P. salmonis antigens, since no cross reaction was detected with Renibacterium salmoninarum and Yersinia ruckeri. The vaccinated and control fish were challenged 60 days after the second dose of DNA with 2.5 × 107 P. salmonis corresponding to 7.5 times the LD50. At 30 days post-challenge, 100{\%} mortality was obtained with the control fish while 20{\%} of the vaccinated animals survived. All surviving fish exhibited a lower bacterial load in the kidney than control fish. The expression library was also tested in Balb/c mice and it was found that the humoral immune response was specific to P. salmonis and it was dependent on the amount of DNA injected.",
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Immunoresponse of Coho salmon immunized with a gene expression library from Piscirickettsia salmonis. / Miquel, Alvaro; Müller, Ilse; Ferrer, Pablo; Valenzuela, Pablo D T; Burzio, Luis O.

En: Biological Research, Vol. 36, N.º 3-4, 01.01.2003, p. 313-323.

Resultado de la investigación: Article

TY - JOUR

T1 - Immunoresponse of Coho salmon immunized with a gene expression library from Piscirickettsia salmonis

AU - Miquel, Alvaro

AU - Müller, Ilse

AU - Ferrer, Pablo

AU - Valenzuela, Pablo D T

AU - Burzio, Luis O.

PY - 2003/1/1

Y1 - 2003/1/1

N2 - We have used the expression library immunization technology to study the protection of Coho salmon Oncorhynchus kisutch to the infection with Piscirickettsia salmonis. Purified DNA from this bacterium was sonicated and the fragments were cloned in the expression vector pCMV-Bios. Two libraries were obtained containing 22,000 and 28,000 colonies and corresponding to approximately 8 and 10 times the genome of the pathogen, respectively. On average, the size of the inserts ranged between 300 and 1,000 bp. The plasmid DNA isolated from one of these libraries was purified and 20 μg were injected intramuscularly into 60 fish followed by a second dose of 10 μg applied 40 days later. As control, fish were injected with the same amount of DNA of the vector pCMV-Bios without insert. The titer of IgM anti-P. salmonis of vaccinated fish, evaluated 60 days post-injection, was significantly higher than that of the control group injected with the vector alone. Moreover, this response was specific against P. salmonis antigens, since no cross reaction was detected with Renibacterium salmoninarum and Yersinia ruckeri. The vaccinated and control fish were challenged 60 days after the second dose of DNA with 2.5 × 107 P. salmonis corresponding to 7.5 times the LD50. At 30 days post-challenge, 100% mortality was obtained with the control fish while 20% of the vaccinated animals survived. All surviving fish exhibited a lower bacterial load in the kidney than control fish. The expression library was also tested in Balb/c mice and it was found that the humoral immune response was specific to P. salmonis and it was dependent on the amount of DNA injected.

AB - We have used the expression library immunization technology to study the protection of Coho salmon Oncorhynchus kisutch to the infection with Piscirickettsia salmonis. Purified DNA from this bacterium was sonicated and the fragments were cloned in the expression vector pCMV-Bios. Two libraries were obtained containing 22,000 and 28,000 colonies and corresponding to approximately 8 and 10 times the genome of the pathogen, respectively. On average, the size of the inserts ranged between 300 and 1,000 bp. The plasmid DNA isolated from one of these libraries was purified and 20 μg were injected intramuscularly into 60 fish followed by a second dose of 10 μg applied 40 days later. As control, fish were injected with the same amount of DNA of the vector pCMV-Bios without insert. The titer of IgM anti-P. salmonis of vaccinated fish, evaluated 60 days post-injection, was significantly higher than that of the control group injected with the vector alone. Moreover, this response was specific against P. salmonis antigens, since no cross reaction was detected with Renibacterium salmoninarum and Yersinia ruckeri. The vaccinated and control fish were challenged 60 days after the second dose of DNA with 2.5 × 107 P. salmonis corresponding to 7.5 times the LD50. At 30 days post-challenge, 100% mortality was obtained with the control fish while 20% of the vaccinated animals survived. All surviving fish exhibited a lower bacterial load in the kidney than control fish. The expression library was also tested in Balb/c mice and it was found that the humoral immune response was specific to P. salmonis and it was dependent on the amount of DNA injected.

KW - Expression library immunization

KW - Genetic vaccine

KW - Piscirickettsia salmonis

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U2 - 10.4067/S0716-97602003000300004

DO - 10.4067/S0716-97602003000300004

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AN - SCOPUS:0242624743

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EP - 323

JO - Biological Research

JF - Biological Research

SN - 0716-9760

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