PUBLICATION
Characterization of myeloid-specific peroxidase, keratin 8, and dual specificity phosphatase 1 as innate immune genes involved in theresistance of crucian carp (Carassius auratus gibelio) to Cyprinid herpesvirus 2 infection
- Authors
- Podok, P., Wang, H., Xu, L., Xu, D., Lu, L.
- ID
- ZDB-PUB-141015-2
- Date
- 2014
- Source
- Fish & shellfish immunology 41(2): 531-40 (Journal)
- Registered Authors
- Wang, Hao
- Keywords
- Crucian carp Carassius auratus gibelio, Cyprinid herpesvirus 2, Dual specificity phosphatase 1, Keratin 8, Myeloid-specific peroxidase
- MeSH Terms
-
- Animals
- Cloning, Molecular
- DNA, Complementary/genetics
- Dual-Specificity Phosphatases/genetics
- Dual-Specificity Phosphatases/metabolism
- Fish Diseases/immunology*
- Fish Diseases/virology*
- Gene Expression Profiling
- Goldfish*
- Herpesviridae Infections/immunology
- Herpesviridae Infections/veterinary*
- Hybridization, Genetic/genetics*
- Immunity, Innate/genetics*
- Keratin-8/genetics
- Keratin-8/metabolism
- Myeloid Cells/metabolism
- Open Reading Frames/genetics
- Peroxidase/genetics
- Peroxidase/metabolism
- Phylogeny
- Real-Time Polymerase Chain Reaction
- Species Specificity
- PubMed
- 25312688 Full text @ Fish Shellfish Immunol.
Citation
Podok, P., Wang, H., Xu, L., Xu, D., Lu, L. (2014) Characterization of myeloid-specific peroxidase, keratin 8, and dual specificity phosphatase 1 as innate immune genes involved in theresistance of crucian carp (Carassius auratus gibelio) to Cyprinid herpesvirus 2 infection. Fish & shellfish immunology. 41(2):531-40.
Abstract
Myeloid-specific peroxidase (MPO), keratin 8 (KRT-8), and dual specificity phosphatase 1 (DUSP-1) are believed to play essential roles in innate immunity. Through suppression subtractive hybridization (SSH) analysis, we previously identified MPO, KRT-8, and DUSP-1 as the three genes that were the most significantly upregulated in crucian carp (Carassius auratus gibelio) that survived Cyprinid herpesvirus 2 (CyHV-2) infection. Here, we have further characterized these three genes and their response to pathogen challenge. The open reading frames (ORF) of MPO, KRT-8, and DUSP-1 were cloned by RACE technique and sequenced. The full-length cDNAs of the three genes contained ORFs of 2,289, 1,575 and 1,083 bp respectively. The polypeptides from each ORF were projected to contain 762 (MPO), 524 (KRT-8), and 360 (DUSP-1) amino acids. Phylogenetic analysis showed that the three genes were most closely related to zebrafish. We found that MPO, KRT-8, and DUSP-1 were expressed at low levels in all of the tissues examined in healthy crucian carp. Quantitative real-time RT-PCR analysis indicated that MPO, KRT-8, and DUSP-1 mRNA expression was significantly upregulated within 72 hours of CyHV-2 infection compared to mock infected controls. Maximum expression of MPO was detected at 24 hpi (2.71-fold, P < 0.05). While, 12 hpi (3.80-fold, P < 0.01) and 6 hpi (8.70-fold, P < 0.01) were the highest expression time points for KRT-8 and DUSP-1, respectively. In contrast, after A. hydrophila challenge, the transcripts of these three genes remained unchanged or slightly down-regulated. For the fish survived from viral infection, expression levels of MPO and KRT-8 were 2.72 fold and 2.47 fold higher than those of fish died from acute infection, and similar level of DUSP-1 was observed in samples of survived fish. These data suggested MPO, KRT-8 and DUSP-1 might be involved in the antiviral, but not antibacterial innate immune response in crucian carp. These findings also support the use of MPO and KRT-8 as immunological markers for a response to viral infection in crucian carp.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping