title: G-protein coupled receptor 18 (GPR18) in channel catfish: expression analysis and efficacy as immunostimulant against Aeromonas hydrophila infection.
abstract: The objectives of this study were: 1) to determine the transcriptional profiles of G-protein coupled receptor 18 (GPR18) in channel catfish after infection with Aeromonas hydrophila compared to that in healthy catfish; 2) to determine whether over-expression of GPR18 in catfish gill cells will offer protection against infection of A. hydrophila; 3) to determine whether recombinant pcDNA-GPR18 could be used as an immunostimulant to protect channel catfish against A. hydrophila infection. Quantitative PCR revealed that the transcription levels of GPR18 in all tissues of infected catfish were significantly (P < 0.05) induced except in the intestine. When pcDNA3.2-vectored recombinant GPR18 was transfected in catfish gill cells G1B, the over-expression of pcDNA-GPR18 offered significant (P < 0.05) protection to G1B cells against A. hydrophila infection. When channel catfish were intraperitoneally injected with QCDCR adjuvant formulated pcDNA-GPR18 and challenged with a highly virulent A. hydrophila strain at 1-, 2-, 14-, and 28-days post treatment, pcDNA-GPR18 offered 50%, 100%, 57%, and 55% protection to channel catfish, respectively. Macrophages of fish treated with pcDNA-GPR18 produced significantly (P < 0.05) higher amounts of reactive oxygen species and nitric oxide than that of fish treated with pcDNA vector alone. In addition, serum lysozyme activity of catfish injected with pcDNA-GPR18 was significantly (P < 0.08) increased. Taken together, our results suggest that pcDNA-GPR18 could be used as a novel immunostimulant to provide immediate protection to channel catfish against A. hydrophila infection.
authors.affiliation: Aquatic Animal Health Research Unit, USDA-ARS, 990 Wire Road, Auburn, AL 36832, USA. Julia.Pridgeon@ars.usda.gov
authors.forename: Julia W
authors.initials: JW
authors.lastname: Pridgeon
authors.forename: Phillip H
authors.initials: PH
authors.lastname: Klesius
mesh_terms: D016980:Aeromonas hydrophila / Q000502:physiology; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D005393:Fish Diseases / Q000276:immunology / Q000382:microbiology; D029941:Fish Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D020869:Gene Expression Profiling / Q000662:veterinary; D005786:Gene Expression Regulation; D005880:Gills / Q000378:metabolism / Q000382:microbiology; D016905:Gram-Negative Bacterial Infections / Q000276:immunology / Q000382:microbiology / Q000662:veterinary; D007059:Ictaluridae / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D008969:Molecular Sequence Data; D009928:Organ Specificity; D010802:Phylogeny; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D016415:Sequence Alignment / Q000662:veterinary
chemical_list: D029941:Fish Proteins; D043562:Receptors, G-Protein-Coupled
keywords: Aeromonas hydrophila; Channel catfish; G-protein coupled receptor 18; Immunostimulant