Codon changed immobilization antigen (iAg), a potent DNA vaccine in fish against Cryptocaryon irritans infection

Highlights ► Codon changed iAg (optiAg) expressed in both prokaryotic and eukaryotic cell systems. ► In vitro immunological data confirmed that optiAg retained the native confirmation. ► iAg is immunogenic due to the protection it conferred to the immunized fish. ► iAg can be a potent DNA vaccine in...

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Published inVaccine Vol. 30; no. 5; pp. 893 - 903
Main Authors Jose Priya, T.A, Lin, Yen-Hong, Wang, Yu-Chi, Yang, Chii-Shen, Chang, Poh-Shing, Song, Yen-Ling
Format Journal Article
LanguageEnglish
Published Netherlands Elsevier Ltd 20.01.2012
Subjects
Allergy and Immunology
amino acids
Animals
antigens
Antigens, Protozoan - genetics
Antigens, Protozoan - immunology
antiserum
Ciliophora - genetics
Ciliophora - immunology
Ciliophora Infections - immunology
Ciliophora Infections - prevention & control
Ciliophora Infections - veterinary
Codon change
Cryptocaryon irritans
DNA vaccine
DNA, Protozoan - chemistry
DNA, Protozoan - genetics
emulsions
enzyme-linked immunosorbent assay
eukaryotic cells
Fish Diseases - immunology
Fish Diseases - prevention & control
grouper
Immobilization antigen
immunization
Injections, Intramuscular
intramuscular injection
Marine
Molecular Sequence Data
plasmids
Protozoa
Protozoan Infections, Animal - immunology
Protozoan Infections, Animal - prevention & control
Protozoan Proteins - genetics
Protozoan Proteins - immunology
Protozoan Vaccines - administration & dosage
Protozoan Vaccines - genetics
Protozoan Vaccines - immunology
recombinant vaccines
Sequence Analysis, DNA
stop codon
Survival Analysis
Vaccines, DNA - administration & dosage
Vaccines, DNA - genetics
Vaccines, DNA - immunology
Western blotting
DNA vaccine
Codon change
Cryptocaryon irritans
Immobilization antigen
Online AccessGet full text

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Abstract Highlights ► Codon changed iAg (optiAg) expressed in both prokaryotic and eukaryotic cell systems. ► In vitro immunological data confirmed that optiAg retained the native confirmation. ► iAg is immunogenic due to the protection it conferred to the immunized fish. ► iAg can be a potent DNA vaccine in fish against infection by C. irritans.
AbstractList The immobilization antigen (iAg) DNA sequence from Chiayi isolate of Cryptocaryon irritans was computationally reviewed to replace the stop codons with suitable amino acids and its GC content was intensified. The plasmid construct comprising the codon changed iAg (optiAg/optimized iAg) was successfully expressed in the bacterial strain BL21 and also in grouper fin cells (GF-1). Results of immobilization assay, ELISA and western blot of C. irritans theront and recombinant iAg by grouper antiserum against optiAg DNA indicated that the codon changed iAg retains the native conformation. The DNA vaccine construct pcDNA3.1-optiAg was encapsulated in water-oil-water triple layer emulsions measuring 19 μm diameters and was used for the immunization experiment. In trial I experiment, grouper fish were immunized twice via intramuscular injection with the pcDNA3.1-optiAg and were challenged with C. irritans at 8-day post immunization (dpi), which resulted in 46% relative percent survival (RPS). In trial II, single immunization with pcDNA3.1-optiAg boosted with recombinant iAg protein, resulted in 40% RPS. The data from this study reveal that codon change in iAg not only accomplished the expression of iAg protein in both prokaryotic and eukaryotic cell systems, but also optiAg was proved as immunogenic due to the protection it confers to the immunized fish against C. irritans infection. Hence, it is concluded that iAg can be a potent DNA vaccine in fish against infection of the ciliated protozoan, C. irritans.
Highlights ► Codon changed iAg (optiAg) expressed in both prokaryotic and eukaryotic cell systems. ► In vitro immunological data confirmed that optiAg retained the native confirmation. ► iAg is immunogenic due to the protection it conferred to the immunized fish. ► iAg can be a potent DNA vaccine in fish against infection by C. irritans.
The immobilization antigen (iAg) DNA sequence from Chiayi isolate of Cryptocaryon irritans was computationally reviewed to replace the stop codons with suitable amino acids and its GC content was intensified. The plasmid construct comprising the codon changed iAg (optiAg/optimized iAg) was successfully expressed in the bacterial strain BL21 and also in grouper fin cells (GF-1). Results of immobilization assay, ELISA and western blot of C. irritans theront and recombinant iAg by grouper antiserum against optiAg DNA indicated that the codon changed iAg retains the native conformation. The DNA vaccine construct pcDNA3.1-optiAg was encapsulated in water-oil-water triple layer emulsions measuring 19 mu m diameters and was used for the immunization experiment. In trial I experiment, grouper fish were immunized twice via intramuscular injection with the pcDNA3.1-optiAg and were challenged with C. irritans at 8-day post immunization (dpi), which resulted in 46% relative percent survival (RPS). In trial II, single immunization with pcDNA3.1-optiAg boosted with recombinant iAg protein, resulted in 40% RPS. The data from this study reveal that codon change in iAg not only accomplished the expression of iAg protein in both prokaryotic and eukaryotic cell systems, but also optiAg was proved as immunogenic due to the protection it confers to the immunized fish against C. irritans infection. Hence, it is concluded that iAg can be a potent DNA vaccine in fish against infection of the ciliated protozoan, C. irritans.
The immobilization antigen (iAg) DNA sequence from Chiayi isolate of Cryptocaryon irritans was computationally reviewed to replace the stop codons with suitable amino acids and its GC content was intensified. The plasmid construct comprising the codon changed iAg (optiAg/optimized iAg) was successfully expressed in the bacterial strain BL21 and also in grouper fin cells (GF-1). Results of immobilization assay, ELISA and western blot of C. irritans theront and recombinant iAg by grouper antiserum against optiAg DNA indicated that the codon changed iAg retains the native conformation. The DNA vaccine construct pcDNA3.1-optiAg was encapsulated in water–oil–water triple layer emulsions measuring 19μm diameters and was used for the immunization experiment. In trial I experiment, grouper fish were immunized twice via intramuscular injection with the pcDNA3.1-optiAg and were challenged with C. irritans at 8-day post immunization (dpi), which resulted in 46% relative percent survival (RPS). In trial II, single immunization with pcDNA3.1-optiAg boosted with recombinant iAg protein, resulted in 40% RPS. The data from this study reveal that codon change in iAg not only accomplished the expression of iAg protein in both prokaryotic and eukaryotic cell systems, but also optiAg was proved as immunogenic due to the protection it confers to the immunized fish against C. irritans infection. Hence, it is concluded that iAg can be a potent DNA vaccine in fish against infection of the ciliated protozoan, C. irritans.
► Codon changed iAg (optiAg) expressed in both prokaryotic and eukaryotic cell systems. ► In vitro immunological data confirmed that optiAg retained the native confirmation. ► iAg is immunogenic due to the protection it conferred to the immunized fish. ► iAg can be a potent DNA vaccine in fish against infection by C. irritans. The immobilization antigen (iAg) DNA sequence from Chiayi isolate of Cryptocaryon irritans was computationally reviewed to replace the stop codons with suitable amino acids and its GC content was intensified. The plasmid construct comprising the codon changed iAg (optiAg/optimized iAg) was successfully expressed in the bacterial strain BL21 and also in grouper fin cells (GF-1). Results of immobilization assay, ELISA and western blot of C. irritans theront and recombinant iAg by grouper antiserum against optiAg DNA indicated that the codon changed iAg retains the native conformation. The DNA vaccine construct pcDNA3.1-optiAg was encapsulated in water–oil–water triple layer emulsions measuring 19μm diameters and was used for the immunization experiment. In trial I experiment, grouper fish were immunized twice via intramuscular injection with the pcDNA3.1-optiAg and were challenged with C. irritans at 8-day post immunization (dpi), which resulted in 46% relative percent survival (RPS). In trial II, single immunization with pcDNA3.1-optiAg boosted with recombinant iAg protein, resulted in 40% RPS. The data from this study reveal that codon change in iAg not only accomplished the expression of iAg protein in both prokaryotic and eukaryotic cell systems, but also optiAg was proved as immunogenic due to the protection it confers to the immunized fish against C. irritans infection. Hence, it is concluded that iAg can be a potent DNA vaccine in fish against infection of the ciliated protozoan, C. irritans.
Author Lin, Yen-Hong
Wang, Yu-Chi
Yang, Chii-Shen
Song, Yen-Ling
Jose Priya, T.A
Chang, Poh-Shing
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Issue 5
Keywords DNA vaccine
Codon change
Cryptocaryon irritans
Immobilization antigen
Language English
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Snippet Highlights ► Codon changed iAg (optiAg) expressed in both prokaryotic and eukaryotic cell systems. ► In vitro immunological data confirmed that optiAg retained...
► Codon changed iAg (optiAg) expressed in both prokaryotic and eukaryotic cell systems. ► In vitro immunological data confirmed that optiAg retained the native...
The immobilization antigen (iAg) DNA sequence from Chiayi isolate of Cryptocaryon irritans was computationally reviewed to replace the stop codons with...
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StartPage 893
SubjectTerms Allergy and Immunology
amino acids
Animals
antigens
Antigens, Protozoan - genetics
Antigens, Protozoan - immunology
antiserum
Ciliophora - genetics
Ciliophora - immunology
Ciliophora Infections - immunology
Ciliophora Infections - prevention & control
Ciliophora Infections - veterinary
Codon change
Cryptocaryon irritans
DNA vaccine
DNA, Protozoan - chemistry
DNA, Protozoan - genetics
emulsions
enzyme-linked immunosorbent assay
eukaryotic cells
Fish Diseases - immunology
Fish Diseases - prevention & control
grouper
Immobilization antigen
immunization
Injections, Intramuscular
intramuscular injection
Marine
Molecular Sequence Data
plasmids
Protozoa
Protozoan Infections, Animal - immunology
Protozoan Infections, Animal - prevention & control
Protozoan Proteins - genetics
Protozoan Proteins - immunology
Protozoan Vaccines - administration & dosage
Protozoan Vaccines - genetics
Protozoan Vaccines - immunology
recombinant vaccines
Sequence Analysis, DNA
stop codon
Survival Analysis
Vaccines, DNA - administration & dosage
Vaccines, DNA - genetics
Vaccines, DNA - immunology
Western blotting
Title Codon changed immobilization antigen (iAg), a potent DNA vaccine in fish against Cryptocaryon irritans infection
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https://dx.doi.org/10.1016/j.vaccine.2011.11.102
https://www.ncbi.nlm.nih.gov/pubmed/22154774
https://search.proquest.com/docview/915633302
https://search.proquest.com/docview/918068779
Volume 30
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