title: Apolipoprotein A1 in channel catfish: transcriptional analysis, antimicrobial activity, and efficacy as plasmid DNA immunostimulant against Aeromonas hydrophila infection.
abstract: The objectives of this study were to: 1) determine transcriptional profiles of apolipoprotein A1 (ApoA1) in collected channel catfish tissues after infection with Aeromonas hydrophila by bath immersion; 2) investigate whether recombinant channel catfish apolipoprotein A1 produced in Escherichia coli expression system possesses any antimicrobial activity against A. hydrophila; 3) evaulate whether recombinant channel catfish apolipoprotein A1 plasmid DNA could be used as immunostimulant to protect fish against A. hydrophila infection. Quantitative PCR revealed that the transcription levels of ApoA1 in infected catfish were significantly (P < 0.05) more induced in the anterior kidney. Recombinant apoA1 produced in E. coli expression system exhibited lytic activity against Gram-positive Micrococcus lysodeikticus and Gram-negative A. hydrophila. When pcDNA3.2-vectored recombinant apoA1 was transfected in channel catfish gill cells G1B, the over-expression of pcDNA-ApoA1 offered significant (P < 0.05) protection to G1B cells against A. hydrophila infection. When channel catfish were intraperitoneally injected with QCDCR adjuvant formulated pcDNA-ApoA1 and challenged with a highly virulent A. hydrophila strain AL-09-71 at two days post injection, pcDNA-ApoA1 injection offered 100% protection to channel catfish. Macrophages of fish injected with pcDNA-ApoA1 produced significantly (P < 0.05) higher amounts of reactive oxygen species and nitric oxide than that of fish injected with pcDNA vector alone. Our results suggest that pcDNA-ApoA1 could be used as a novel immunostimulant to offer immediate protection to catfish against A. hydrophila infection.
authors: Aquatic Animal Health Research Unit, USDA-ARS, 990 Wire Road, Auburn, AL 36832, USA. Julia.Pridgeon@ars.usda.gov
authors: Julia W
authors: JW
authors: Pridgeon
authors: Phillip H
authors: PH
authors: Klesius
mesh_terms: D000276:Adjuvants, Immunologic; D016980:Aeromonas hydrophila / Q000276:immunology*; D000595:Amino Acid Sequence; D000818:Animals; D000890:Anti-Infective Agents / Q000378:metabolism; D016632:Apolipoprotein A-I / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D001428:Bacterial Vaccines / Q000627:therapeutic use*; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000378:metabolism; D018076:DNA, Complementary; D004926:Escherichia coli / Q000235:genetics; D005393:Fish Diseases / Q000235:genetics / Q000517:prevention & control*; D029941:Fish Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation; D016905:Gram-Negative Bacterial Infections / Q000235:genetics / Q000517:prevention & control / Q000662:veterinary*; D007059:Ictaluridae / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D009928:Organ Specificity; D010802:Phylogeny; D010957:Plasmids / Q000378:metabolism; D012313:RNA; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment / Q000662:veterinary
chemical_list: D000276:Adjuvants, Immunologic; D000890:Anti-Infective Agents; D016632:Apolipoprotein A-I; D001428:Bacterial Vaccines; D004269:DNA, Bacterial; D018076:DNA, Complementary; D029941:Fish Proteins; D011994:Recombinant Proteins; D012313:RNA
keywords: Aeromonas hydrophila; Apolipoprotein A1; Channel catfish; Immunostimulant