Characterization of a Novel Cysteine Protease Inhibitor from Poultry Red Mites: Potential Vaccine for Chickens.
Ontology highlight
ABSTRACT:
SUBMITTER: Fujisawa S
PROVIDER: S-EPMC8706574 | biostudies-literature |
REPOSITORIES: biostudies-literature
Similar Datasets
Project description:Poultry red mites (PRMs, Dermanyssus gallinae) are ectoparasites that negatively affect farmed chickens, leading to serious economic losses worldwide. Acaricides have been used to control PRMs in poultry houses. However, some PRMs have developed resistance to acaricides, and therefore different approaches are required to manage the problems caused by PRMs. Vaccination of chickens is one of the methods being considered to reduce the number of PRMs in poultry houses. In a previous study, a cysteine protease, Deg-CPR-1, was identified as a candidate vaccine against PRMs distributed in Europe. In this study, we investigated the characteristics of Deg-CPR-1. A phylogenetic analysis revealed that Deg-CPR-1 is closely related to the digestive cysteine proteases of other mite species, and it was classified into a cluster different from that of chicken cathepsins. Deg-CPR-1 of PRMs in Japan has an amino acid substitution compared with that of PRMs in Europe, but it showed efficacy as a vaccine, consistent with previous findings. Deg-CPR-1 exhibited cathepsin L-like enzyme activity. In addition, the Deg-CPR-1 mRNA was expressed in the midgut and in all stages of PRMs that feed on blood. These results imply that Deg-CPR-1 in the midgut may have important functions in physiological processes, and the inhibition of its expression may contribute to the efficacy of a Deg-CPR-1-based vaccine. Further research is required to fully understand the mechanisms of vaccine efficacy.
| S-EPMC7863971 | biostudies-literature
Project description:Poultry red mites (PRMs, Dermanyssus gallinae) are hematophagous ectoparasites that negatively affect egg production, which causes serious economic losses to the poultry industry worldwide. Currently, the emergence of acaricide-resistant PRMs has impeded PRM control in poultry farms. Several alternatives for acaricide use have been described for managing PRM-caused problems. Vaccination is among the methods for controlling PRMs in poultry houses. Currently, several candidates for vaccine antigens have been identified. This study identified a cysteine protease, Deg-CPR-2, which differs from 2 other previously reported cysteine proteases in PRMs, from previously obtained data from RNA-sequencing (RNA-seq) analysis. We investigated the characteristics of Deg-CPR-2 and assessed its efficacy as a vaccine antigen in vitro. Phylogenetic analysis revealed that Deg-CPR-2 belonged to a different cluster from those of other cysteine proteases in PRMs. This cluster also included cathepsin L-like proteases, enzymes thought to be involved in hemoglobin digestion in ticks. Expression analysis revealed Deg-CPR-2 expression in midguts and all the life-stages; however, there were differences in the expression levels across the life-stages. The enzyme activity of recombinant Deg-CPR-2 was inhibited in the presence of a cysteine protease inhibitor, which suggests that Deg-CPR-2 functions as a cysteine protease in PRMs. Finally, there was an in vitro increase in the mortality of PRMs, mainly protonymphs that were artificially fed with plasma from chickens immunized with Deg-CPR-2. These findings suggest that Deg-CPR-2 may contribute to protein digestion in the midgut of PRMs and is crucially involved in physiological processes in PRMs. Additionally, immunization with Deg-CPR-2 may reduce the number of protonymphs, and Deg-CPR-2 should be considered as a candidate antigen for anti-PRM vaccine development.
| S-EPMC8743220 | biostudies-literature
Project description:IntroductionPoultry red mites (PRMs, Dermanyssus gallinae), blood-sucking ectoparasites, are a threat to the poultry industry because of reduced production caused by infestation. In addition, tropical fowl mites (TFMs, Ornithonyssus bursa) and northern fowl mites (NFMs, Ornithonyssus sylviarum) are hematophagous, distributed in various regions, genetically and morphologically close to PRMs, and cause similar problems to the poultry industry. Vaccine approaches have been studied for PRM control, and several molecules have been identified in PRMs as candidates for effective vaccine antigens. The development of an anti-PRM vaccine as a universal vaccine with broad efficacy against avian mites could improve the productivity of poultry farms worldwide. Molecules that are highly conserved among avian mites and have critical functions in the physiology and growth of mites could be ideal antigen candidates for the development of universal vaccines. Ferritin 2 (FER2), an iron-binding protein, is critical for the reproduction and survival of PRMs and has been reported as a useful vaccine antigen for the control of PRMs and a candidate for the universal vaccine antigen in some tick species.Method and resultsHerein, we identified and characterized FER2 in TFMs and NFM. Compared with the sequence of PRM, the ferroxidase centers of the heavy chain subunits were conserved in FER2 of TFMs and NFMs. Phylogenetic analysis revealed that FER2 belongs to clusters of secretory ferritins of mites and other arthropods. Recombinant FER2 (rFER2) proteins from PRMs, TFMs, and NFMs exhibited iron-binding abilities. Immunization with each rFER2 induced strong antibody responses in chickens, and each immune plasma cross-reacted with rFER2 from different mites. Moreover, mortality rates of PRMs fed with immune plasma against rFER2 from TFMs or NFMs, in addition to PRMs, were higher than those of control plasma.DiscussionrFER2 from each avian mite exhibited anti-PRM effects. This data suggests that it has the potential to be used as an antigen candidate for a universal vaccine against avian mites. Further studies are needed to access the usefulness of FER2 as a universal vaccine for the control of avian mites.
| S-EPMC10149675 | biostudies-literature
Project description:To assess their potential to control poultry red mites (Dermanyssus gallinae), we tested selected predaceous mites (Androlaelaps casalis and Stratiolaelaps scimitus) that occur naturally in wild bird nests or sometimes spontaneously invade poultry houses. This was done under laboratory conditions in cages, each with 2-3 laying hens, initially 300 poultry red mites and later the release of 1,000 predators. These small-scale tests were designed to prevent mite escape from the cages and they were carried out in three replicates at each of three temperature regimes: 26, 30 (constant day and night) and 33-25 °C (day-night cycle). After 6 weeks total population sizes of poultry red mites and predatory mites were assessed. For the temperature regimes of 26 and 33/25 °C S. scimitus reduced the poultry red mite population relative to the control experiments by a factor 3 and 30, respectively, and A. casalis by a factor of 18 and 55, respectively. At 30 °C the predators had less effect on red mites, with a reduction of 1.3-fold for S. scimitus and 5.6-fold for A. casalis. This possibly reflected hen manure condition or an effect of other invertebrates in the hen feed. Poultry red mite control was not negatively affected by temperatures as high as 33 °C and was always better in trials with A. casalis than in those with S. scimitus. In none of the experiments predators managed to eradicate the population of poultry red mites. This may be due to a prey refuge effect since most predatory mites were found in and around the manure tray at the bottom of the cage, whereas most poultry red mites were found higher up in the cage (i.e. on the walls, the cover, the perch, the nest box and the food box). The efficacy of applying predatory mites in the poultry industry may be promoted by reducing this refuge effect, boosting predatory mite populations using alternative prey and prolonged predator release devices. Biocontrol success, however, will strongly depend on how the poultry is housed in practice (free range, cage or aviary systems) and on which chemicals are applied to disinfect poultry houses and to control other pests.
| S-EPMC3487000 | biostudies-literature
Project description:Additional biomarkers for the development and progression of colorectal cancer (CRC) remain to be identified. Hence, the current study aimed to identify potential diagnostic markers for CRC. Analyses of cysteine protease inhibitor [cystatins (CSTs)] expression in CRC samples and its correlation with cancer stage or survival in patients with CRC demonstrated that CRC tissues had greater CST1 and CST2 mRNA expression compared to noncancerous adjacent tissues, while higher CST2 mRNA expression in CRC tissues was correlated with advanced stages and disease-free survival in patients with CRC, encouraging further exploration on the role of CST2 in CRC. Through an online database search and tissue microarray (TMA), we confirmed that CRC samples had higher CST2 expression compared to noncancerous adjacent tissue or normal colorectal tissues at both the mRNA and protein levels. TMA also revealed that colorectal adenoma, CRC, and metastatic CRC tissues exhibited a significantly increased CST2 protein expression. Accordingly, survival analysis demonstrated that the increase in CST2 protein expression was correlated with shorter overall survival of patients with CRC. Moreover, our results found a significant upregulation of CST2 in multiple cancer tissues. Taken together, these findings suggest the potential role of CST2 as a diagnostic and prognostic biomarker for CRC.
| S-EPMC8317524 | biostudies-literature
Project description:Poultry waste is an abundant renewable source for the recovery of several value-added metabolites with potential industrial applications. This study describes the production of protease on poultry waste, with the subsequent use of the same poultry waste for the extraction of antioxidants. An extracellular protease-producing strain was isolated from Cuddalore coast, India, and identified as Streptomyces sp. MAB18. Its protease was purified 17.13-fold with 21.62% yield with a specific activity of 2398.36?U/mg and the molecular weight was estimated as 43?kDa. The enzyme was optimally active at pH 8-10 and temperature 50-60 ° C and it was most stable up to pH 12 and 6-12% of NaCl concentration. The enzyme activity was reduced when treated with Hg(2+), Pb(2+), and SDS and stimulated by Fe(2+), Mg(2+), Triton X-100, DMSO (dimethyl sulfoxide), sodium sulphite, and ?-mercaptoethanol. Furthermore, the antioxidant activities of protease were evaluated using in vitro antioxidant assays, such as DPPH radical-scavenging activity, O2 scavenging activity, NO scavenging activity, Fe(2+) chelating activity, and reducing power. The enzyme showed important antioxidant potential with an IC50 value of 78 ± 0.28?mg/mL. Results of the present study indicate that the poultry waste-derived protease may be useful as supplementary protein and antioxidant in the animal feed formulations.
| S-EPMC3749541 | biostudies-literature
Project description:Protease inhibitors are involved in the regulation of endogenous cysteine proteases during seed development and play a defensive role because of their ability to inhibit exogenous proteases such as those present in the digestive tracts of insects. Araucaria angustifolia seeds, which can be used in human and animal feed, were investigated for their potential for the development of agricultural biotechnology and in the field of human health. In the pine nuts extract, which blocked the activities of cysteine proteases, it was detected potent insecticidal activity against termites (Nasutitermes corniger) belonging to the most abundant termite genus in tropical regions. The cysteine inhibitor (AaCI-2S) was purified by ion-exchange, size exclusion, and reversed-phase chromatography. Its functional and structural stability was confirmed by spectroscopic and circular dichroism studies, and by detection of inhibitory activity at different temperatures and pH values. Besides having activity on cysteine proteases from C. maculatus digestive tract, AaCI-2S inhibited papain, bromelain, ficin, and cathepsin L and impaired cell proliferation in gastric and prostate cancer cell lines. These properties qualify A. angustifolia seeds as a protein source with value properties of natural insecticide and to contain a protease inhibitor with the potential to be a bioactive molecule on different cancer cells.
| S-EPMC7760129 | biostudies-literature
Project description:Cathepsin L (CatL) is a lysosomal cysteine protease primarily involved in the terminal degradation of intracellular and endocytosed proteins. More specifically, in humans, CatL has been implicated in cancer progression and metastasis, as well as coronary artery diseases and others. Given this, the search for potent CatL inhibitors is of great importance. In the search for new molecules to perform proteolytic activity regulation, salivary secretions from hematophagous animals have been an important source, as they present protease inhibitors that evolved to disable host proteases. Based on the transcriptome of the Haementeria vizzotoi leech, the cDNA of Cystatin-Hv was selected for this study. Cystatin-Hv was expressed in Pichia pastoris and purified by two chromatographic steps. The kinetic results using human CatL indicated that Cystatin-Hv, in its recombinant form, is a potent inhibitor of this protease, with a Ki value of 7.9 nM. Consequently, the present study describes, for the first time, the attainment and the biochemical characterization of a recombinant cystatin from leeches as a potent CatL inhibitor. While searching out for new molecules of therapeutic interest, this leech cystatin opens up possibilities for the future use of this molecule in studies involving cellular and in vivo models.
| S-EPMC8705320 | biostudies-literature
Project description:BACKGROUND:Sparganosis is a neglected but important food-borne parasitic zoonosis. Clinical diagnosis of sparganosis is difficult because there are no specific manifestations. ELISA using plerocercoid crude or excretory-secretory (ES) antigens has high sensitivity but has cross-reactions with other helminthiases. The aim of this study was to characterize Spirometra erinaceieuropaei cysteine protease (SeCP) and to evaluate its potential application for serodiagnosis of sparganosis. METHODOLOGY/PRINCIPAL FINDINGS:The full length SeCP gene was cloned, and recombinant SeCP (rSeCP) was expressed and purified. Western blotting showed that rSeCP was recognized by the serum of sparganum-infected mice, and anti-rSeCP serum recognized the native SeCP protein of plerocercoid crude or ES antigens. Expression of SeCP was observed at plerocercoid stages but not at the adult and egg stages. Immunolocalization identified SeCP in plerocercoid tegument and parenchymal tissue. The rSeCP had CP activity, and the optimum pH and temperature were 5.5 and 37°C, respectively. Enzymatic activity was significantly inhibited by E-64. rSeCP functions to degrade different proteins and the function was inhibited by anti-rSeCP serum and E-64. Immunization of mice with rSeCP induced Th2-predominant immune responses and anti-rSeCP antibodies had the potential capabilities to kill plerocercoids in an ADCC assay. The sensitivity of rSeCP-ELISA and ES antigen ELISA was 100% when performed on sera of patients with sparganosis. The specificity of rSeCP-ELISA and ES antigen ELISA was 98.22% (166/169) and 87.57% (148/169), respectively (P<0.05). CONCLUSIONS:The rSeCP had the CP enzymatic activity and SeCP seems to be important for the survival of plerocercoids in host. The rSeCP is a potential diagnostic antigen for sparganosis.
| S-EPMC4457932 | biostudies-literature
Project description:Infestation with poultry red mites (PRM, Dermanyssus gallinae) causes anemia, reduced egg production, and death in serious cases, resulting in significant economic losses to the poultry industry. As a novel strategy for controlling PRMs, vaccine approaches have been focused upon and several candidate vaccine antigens against PRMs have been reported. Tropical (TFM, Ornithonyssus bursa) and northern (NFM, Ornithonyssus sylviarum) fowl mites are also hematophagous and cause poultry industry problems similar to those caused by PRM. Therefore, ideal antigens for anti-PRM vaccines are molecules that cross-react with TFMs and NFMs, producing pesticidal effects similar to those against PRMs. In this study, to investigate the potential feasibility of developing vaccines with broad efficacy across mite species, we identified and characterized cysteine proteases (CPs) of TFMs and NFMs, which were previously reported to be effective vaccine antigens of PRMs. The open reading frames of CPs from TFMs and NFMs had the same sequences, which was 73.0% similar to that of PRMs. Phylogenetic analysis revealed that the CPs of TFMs and NFMs clustered in the same clade as CPs of PRMs. To assess protein functionality, we generated recombinant peptidase domains of CPs (rCP-PDs), revealing all rCP-PDs showed CP-like activities. Importantly, the plasma obtained from chickens immunized with each rCP-PD cross-reacted with rCP-PDs of different mites. Finally, all immune plasma of rCP-PDs reduced the survival rate of PRMs, even when the plasma was collected from chickens immunized with rCP-PDs derived from TFM and NFM. Therefore, CP antigen is a promising, broadly efficacious vaccine candidate against different avian mites.
| S-EPMC10343161 | biostudies-literature