The acute stresses role of the atypical 2-cys peroxiredoxin PmPrx5 in black tiger shrimp (Penaeus monodon) from biological immunity and environmental toxicity stress.

As a unique atypical 2-Cys Peroxiredoxin (Prx) of the Prx-like superfamily, Peroxiredoxin5 (Prx5) possesses special properties, such as its enzymatic mechanism, wide subcellular distribution and high affinity for peroxides and peroxynitrite. Prx5 plays a crucial role in oxidative stress, immune responses, cell apoptosis, proliferation, differentiation, intracellular signaling, the modulation of gene expression, ecdysis, etc. In this paper, we obtained a full-length Prx5 cDNA sequence (designated PmPrx5) from black tiger shrimp (P. monodon). The full-length PmPrx5 cDNA sequence was 1686 bp containing a 5' untranslated region (UTR) of 76 bp with two nucleotide sequences (AAA), a 3' UTR of 1040 bp with a poly (A) tail and two canonical polyadenylation signal sequences (AATAAA), and an open reading frame of 570 bp encoding 189 amino acid residues with a predicted molecular mass of 20 kDa and a theoretical isoelectric point of 6.29. Phylogenetic trees and multiple sequence alignment showed that the PmPrx5 had strong homology with Prx5 proteins from other species, such as similarity with Palaemon carinicauda (69%) and Macrobrachium rosenbergii (69%), containing the highly conserved functional domain. PmPrx5 mRNA was ubiquitously detected in all tested tissues. After P. monodon was exposed to pathogenic bacteria, osmotic pressure, acidity and alkalinity and the heavy metal, the mRNA expression of PmPrx5 in the gills and hepatopancreas was significantly enhanced (P < 0.01) because of the immune response and declined with heavy metal copper and cadmium challenges as time progressed. The recombinant PmPrx5 protein purified in E. coli (DE3) was further confirmed to exhibit antioxidant activity and antibacterial properties to a certain extent using a bacterial growth inhibition test in both liquid and solid cultures in vitro. E. coli transformed with pRSET-PmPrx5 were dramatically protected in response to metal toxicity stress. Thus, PmPrx5 may be developed as a potential therapeutic drug against pathogenic bacteria and as a biomarker for pollutant levels. This work offers useful clues to further explore the functional mechanism of Prx5 in marine shrimp immunity.

Molecular characterization and expression analysis of six peroxiredoxin paralogous genes in gilthead sea bream (Sparus aurata): insights from fish exposed to dietary, pathogen and confinement stressors.

The aim of this work was to underline the physiological role of the antioxidant peroxiredoxin (PRDX) family in gilthead sea bream (Sparus aurata L.), a perciform fish extensively cultured in the Mediterranean area. First, extensive BLAST searches were done on the gilthead sea bream cDNA database of the AQUAMAX European Project (www.sigenae.org/iats), and six contigs were unequivocally identified as PRDX1-6 after sequence completion by RT-PCR. The phylogenetic analysis evidenced three major clades corresponding to PRDX1-4 (true 2-Cyst PRDX subclass), PRDX5 (atypical 2-Cys PRDX subclass) and PRDX6 (1-Cys PRDX subclass) that reflected the present hierarchy of vertebrates. However, the PRDX2 branch of modern fish including gilthead sea bream was related to the monophyletic PRDX1 node rather than to PRDX2 cluster of mammals and primitive fish, which probably denotes the acquisition of novel functions through vertebrate evolution. Transcriptional studies by means of quantitative real-time PCR evidenced a ubiquitous PRDX gene expression that was tissue specific for each PRDX isoform. In a second set of transcriptional studies, liver and head kidney were chosen as target tissues in fish challenged with i) the intestinal parasite Enteromyxum leei, ii) a plant oil (VO) diet with deficiencies in essential fatty acids and iii) prolonged exposure to high-rearing densities. These studies showed that PRDX genes were highly and mostly constitutively expressed in the liver and were not affected by dietary intervention or high density. In contrast, head kidney was highly sensitive to the different experimental challenges: significantly lower values were found for PRDX5 in the three trials, for PRDX6 in parasitized and high density fish and for PRDX1 in parasitized and VO fish. PRDX2, 3 and 5 were decreased only in VO, high density and parasitized animals, respectively. These findings would highlight the role of PRDXs as integrative and highly predictive biomarkers of health and welfare in fish and gilthead sea bream in particular.

Peroxiredoxin I is a negative regulator of Th2-dominant allergic asthma.

Peroxiredoxin (Prx) I, a ubiquitous antioxidant enzyme, is known to protect against inflammation; however, its role in the allergic inflammation remains unidentified. We determined whether intristic Prx I protects against allergic asthma traits using Prx-I knockout (-/-) mice. Prx I (-/-) and wild-type (WT) mice were immunized with ovalbumin (OVA) plus aluminum potassium sulfate (Alum: Th2 adjuvant) and subsequently challenged with OVA. Twenty-four hours after the last OVA challenge, leukocyte influx including eosinophils into bronchoalveolar lavage fluid was significantly greater in Prx I (-/-) mice compared to that in WT mice. On the other hand, when these mice were immunized with OVA+complete Freund's adjuvant (Th1 adjuvant), opposite phenomenon was observed. In the presence of OVA/Alum, peribronchial inflammatory leukocyte infiltration, cholinergic airway resistance, and the lung expression of interleukin (IL)-2 were significantly greater and that of interferon-gamma was significantly lesser in Prx I (-/-) than in WT mice. In vitro, OVA/Alum-sensitized Prx I (-/-) T cells proliferated more profoundly than WT T cells when they were cocultured with syngeneic bone marrow-generated dendritic cells. These results indicate that endogenous Prx I protects against allergen-related Th2-type airway inflammation and hyperresponsiveness, at least partly, via the suppression of the lung expression of IL-2 and regulation of the Th1/Th2 balance in addition to its antioxidative properties. Furthermore, Prx I can inhibit allergen-specific T-cell proliferation through immunological synapse. Our findings implicate an alternative therapeutic value of Prx I in the treatment of Th2-skewed allergic airway inflammatory diseases such as atopic asthma.