PUBLICATION
Molecular characterization of an interleukin-4/13B homolog in grass carp (Ctenopharyngodon idella) and its role in fish against Aeromonas hydrophila infection
- Authors
- Yang, Z.J., Li, C.H., Chen, J., Zhang, H., Li, M.Y., Chen, J.
- ID
- ZDB-PUB-160823-3
- Date
- 2016
- Source
- Fish & shellfish immunology 57: 136-47 (Journal)
- Registered Authors
- Keywords
- Aeromonas hydrophila infection, Bacterial load, Grass carp, Interleukin-4/13B, M2 polarization, Monocyte/macrophage, Sequence analysis, Survival rate
- MeSH Terms
-
- Aeromonas hydrophila/physiology
- Amino Acid Sequence
- Animals
- Bacterial Load*
- Carps*
- Cloning, Molecular
- DNA, Complementary/genetics
- DNA, Complementary/metabolism
- Fish Diseases/genetics
- Fish Diseases/immunology
- Fish Diseases/microbiology
- Fish Proteins/chemistry
- Fish Proteins/genetics*
- Fish Proteins/metabolism
- Gram-Negative Bacterial Infections/genetics
- Gram-Negative Bacterial Infections/immunology
- Gram-Negative Bacterial Infections/microbiology
- Gram-Negative Bacterial Infections/veterinary*
- Immunity, Innate*
- Interleukin-4/chemistry
- Interleukin-4/genetics*
- Interleukin-4/metabolism
- Phylogeny
- RNA, Messenger/genetics
- RNA, Messenger/metabolism
- Recombinant Proteins/chemistry
- Recombinant Proteins/genetics
- Recombinant Proteins/metabolism
- Sequence Alignment/veterinary
- PubMed
- 27546554 Full text @ Fish Shellfish Immunol.
Citation
Yang, Z.J., Li, C.H., Chen, J., Zhang, H., Li, M.Y., Chen, J. (2016) Molecular characterization of an interleukin-4/13B homolog in grass carp (Ctenopharyngodon idella) and its role in fish against Aeromonas hydrophila infection. Fish & shellfish immunology. 57:136-47.
Abstract
Mammalian interleukin 4 (IL-4) and interleukin 13 (IL-13) molecules are anti-inflammatory cytokines mediating the alternative activation of macrophages. However, the role of fish IL-4/13 homologs in monocytes/macrophages (MO/MФ) polarization remains unclear. In this study, we have functionally identified an IL-4/13B homolog in grass carp (Ctenopharyngodon idella), which is termed as CiIL-4/13B. Multiple alignment showed that CiIL-4/13B shared the typical characteristics and structure with other known fish IL-4/13. Phylogenetic analysis showed that CiIL-4/13B is evolutionarily closely related to zebrafish (Danio rerio) and common carp (Cyprinus carpio) IL-4/13B. CiIL-4/13B mRNA was constitutively expressed in tissues and peripheral blood lymphocytes (PBLs) examined, with its highest expression seen in PBLs. Following Aeromonas hydrophila infection, CiIL-4/13B mRNA expression was upregulated. Recombinant CiIL-4/13B (rCiIL-4/13B) was overexpressed in Escherichia coli and purified for a functional study. Using prepared anti-rCiIL-4/13B antiserum, Western blot analysis showed that native CiIL-4/13B in grass carp plasma is N-glycosylated. Intraperitoneal injection of bioactive rCiIL-4/13B significantly increased the survival rate of grass carp against A. hydrophila, and decreased the tissue bacterial load, with a higher dose having better effects. Bioactive rCiIL-4/13B treatment decreased nitrite production and mRNA expression of proinflammatory cytokines (IL-1β and TNF-α), while it increased arginase activity and mRNA expression of anti-inflammatory cytokines (TGF-β and IL-10). The phagocytosis by grass carp MO/MФ had no significant changes by the 8 h treatment of bioactive rCiIL-4/13B compared to that of the negative control, while it was significantly inhibited by the 24 h treatment of bioactive rCiIL-4/13B. The inhibitory effect of rCiIL-4/13B on MO/MФ phagocytosis may be a consequence of MO/MФ proliferation. In summary, our results suggest that CiIL-4/13B plays a protective effect in grass carp against A. hydrophila by inducing alternatively activated MO/MФ.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping