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Characterization of a Trichinella spiralis aminopeptidase and its participation in invasion, development and fecundity
A Trichinella spiralis aminopeptidase (TsAP) has been identified in intestinal infectious larvae (IIL) and adult worms (AW), but its biological function in the T. spiralis life cycle is unknown. The aim of this study was to characterize TsAP and ascertain its functions in the invasion, development a...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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BioMed Central
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7296678/ https://www.ncbi.nlm.nih.gov/pubmed/32539772 http://dx.doi.org/10.1186/s13567-020-00805-w |
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author | Guo, Kai Xia Bai, Ying Ren, Hua Nan Sun, Xiang Yuan Song, Yan Yan Liu, Ruo Dan Long, Shao Rong Zhang, Xi Jiang, Peng Wang, Zhong Quan Cui, Jing |
author_facet | Guo, Kai Xia Bai, Ying Ren, Hua Nan Sun, Xiang Yuan Song, Yan Yan Liu, Ruo Dan Long, Shao Rong Zhang, Xi Jiang, Peng Wang, Zhong Quan Cui, Jing |
author_sort | Guo, Kai Xia |
collection | PubMed |
description | A Trichinella spiralis aminopeptidase (TsAP) has been identified in intestinal infectious larvae (IIL) and adult worms (AW), but its biological function in the T. spiralis life cycle is unknown. The aim of this study was to characterize TsAP and ascertain its functions in the invasion, development and fecundity of T. spiralis. Recombinant TsAP (rTsAP) was expressed and purified. rTsAP has strong immunogenicity. qPCR and western blotting show that TsAP was transcribed and expressed at all T. spiralis lifecycle stages, but the expression level of TsAP mRNA and proteins at IIL and AW stages was obviously higher than those in muscle larvae (ML) and newborn larvae (NBL). The IFT results reveal that TsAP was principally located at the cuticle and the intrauterine embryos of this nematode. rTsAP had the enzymatic activity of natural aminopeptidase to hydrolyze the substrate Leu-pNA with an optimal temperature of 50 °C and optimal pH of 8.0. rTsAP promoted the larval penetration into intestinal epithelial cells, whereas anti-rTsAP antibodies suppressed the larval intrusion; the promotion and suppression was dose-dependently related to rTsAP or anti-rTsAP antibodies. TsAP protein expression level and enzymatic activity were reduced by 50.90 and 49.72% through silencing of the TsAP gene by specific siRNA 842. Intestinal AW and muscle larval burdens, worm length and female reproductive capacity were significantly declined in mice infected with siRNA-transfected ML compared to the control siRNA and PBS group. These results indicate that TsAP participates in the invasion, development and fecundity of T. spiralis and it might be a candidate target for anti-Trichinella vaccines. |
format | Online Article Text |
id | pubmed-7296678 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-72966782020-06-16 Characterization of a Trichinella spiralis aminopeptidase and its participation in invasion, development and fecundity Guo, Kai Xia Bai, Ying Ren, Hua Nan Sun, Xiang Yuan Song, Yan Yan Liu, Ruo Dan Long, Shao Rong Zhang, Xi Jiang, Peng Wang, Zhong Quan Cui, Jing Vet Res Research Article A Trichinella spiralis aminopeptidase (TsAP) has been identified in intestinal infectious larvae (IIL) and adult worms (AW), but its biological function in the T. spiralis life cycle is unknown. The aim of this study was to characterize TsAP and ascertain its functions in the invasion, development and fecundity of T. spiralis. Recombinant TsAP (rTsAP) was expressed and purified. rTsAP has strong immunogenicity. qPCR and western blotting show that TsAP was transcribed and expressed at all T. spiralis lifecycle stages, but the expression level of TsAP mRNA and proteins at IIL and AW stages was obviously higher than those in muscle larvae (ML) and newborn larvae (NBL). The IFT results reveal that TsAP was principally located at the cuticle and the intrauterine embryos of this nematode. rTsAP had the enzymatic activity of natural aminopeptidase to hydrolyze the substrate Leu-pNA with an optimal temperature of 50 °C and optimal pH of 8.0. rTsAP promoted the larval penetration into intestinal epithelial cells, whereas anti-rTsAP antibodies suppressed the larval intrusion; the promotion and suppression was dose-dependently related to rTsAP or anti-rTsAP antibodies. TsAP protein expression level and enzymatic activity were reduced by 50.90 and 49.72% through silencing of the TsAP gene by specific siRNA 842. Intestinal AW and muscle larval burdens, worm length and female reproductive capacity were significantly declined in mice infected with siRNA-transfected ML compared to the control siRNA and PBS group. These results indicate that TsAP participates in the invasion, development and fecundity of T. spiralis and it might be a candidate target for anti-Trichinella vaccines. BioMed Central 2020-06-15 2020 /pmc/articles/PMC7296678/ /pubmed/32539772 http://dx.doi.org/10.1186/s13567-020-00805-w Text en © The Author(s) 2020 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Article Guo, Kai Xia Bai, Ying Ren, Hua Nan Sun, Xiang Yuan Song, Yan Yan Liu, Ruo Dan Long, Shao Rong Zhang, Xi Jiang, Peng Wang, Zhong Quan Cui, Jing Characterization of a Trichinella spiralis aminopeptidase and its participation in invasion, development and fecundity |
title | Characterization of a Trichinella spiralis aminopeptidase and its participation in invasion, development and fecundity |
title_full | Characterization of a Trichinella spiralis aminopeptidase and its participation in invasion, development and fecundity |
title_fullStr | Characterization of a Trichinella spiralis aminopeptidase and its participation in invasion, development and fecundity |
title_full_unstemmed | Characterization of a Trichinella spiralis aminopeptidase and its participation in invasion, development and fecundity |
title_short | Characterization of a Trichinella spiralis aminopeptidase and its participation in invasion, development and fecundity |
title_sort | characterization of a trichinella spiralis aminopeptidase and its participation in invasion, development and fecundity |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7296678/ https://www.ncbi.nlm.nih.gov/pubmed/32539772 http://dx.doi.org/10.1186/s13567-020-00805-w |
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