Cargando…
Molecular identification of Trichinella spiralis nudix hydrolase and its induced protective immunity against trichinellosis in BALB/c mice
BACKGROUND: Nudix hydrolases (Nd) is a widespread superfamily, which is found in all classes of organism, hydrolyse a wide range of organic pyrophosphates and has a ‘housecleaning’ function. The previous study showed that Trichinella spiralis Nd (TsNd) bound to intestinal epithelial cells (IECs), an...
Autores principales: | , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2014
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4287505/ https://www.ncbi.nlm.nih.gov/pubmed/25522912 http://dx.doi.org/10.1186/s13071-014-0600-9 |
_version_ | 1782351799273914368 |
---|---|
author | Long, Shao Rong Wang, Zhong Quan Liu, Ruo Dan Liu, Li Na Li, Ling Ge Jiang, Peng Zhang, Xi Zhang, Zi Fang Shi, Hai Ning Cui, Jing |
author_facet | Long, Shao Rong Wang, Zhong Quan Liu, Ruo Dan Liu, Li Na Li, Ling Ge Jiang, Peng Zhang, Xi Zhang, Zi Fang Shi, Hai Ning Cui, Jing |
author_sort | Long, Shao Rong |
collection | PubMed |
description | BACKGROUND: Nudix hydrolases (Nd) is a widespread superfamily, which is found in all classes of organism, hydrolyse a wide range of organic pyrophosphates and has a ‘housecleaning’ function. The previous study showed that Trichinella spiralis Nd (TsNd) bound to intestinal epithelial cells (IECs), and the vaccination of mice with T7 phage-displayed TsNd polypeptides produced protective immunity. The aim of this study was to clone, express and identify the full-length TsNd and to investigate its immune protection against T. spiralis infection. METHODS: The full-length cDNA sequence of TsNd gene encoding a 46 kDa protein from T. spiralis intestinal infective larvae (IIL) was cloned and identified. The antigenicity of rTsNd was analyzed by Western blot. Transcription and expression of TsNd at T. spiralis different stages were observed by RT-PCR and IFT. The levels of the specific total IgG, IgG1 and IgG2a antibodies to rTsNd were determined by ELISA. The immune protection of rTsNd against T. spiralis infection was investigated. RESULTS: Sequence and phylogenetic analysis revealed that TsNd had a nudix motif located at 226-244aa, which had high homology and the closest evolutionary status with T. pseudospiralis. The rTsNd was obtained after expression and purification. Western blot analysis showed that anti-rTsNd serum recognized the native TsNd protein in crude antigens of muscle larvae (ML), IIL, adult worms (AW) and newborn larvae (NBL), and ES antigens of ML. Transcription and expression of TsNd gene was observed in all developmental stages of T. spiralis (ML, IIL, AW and NBL), with high level expression in IIL. An immunolocalization analysis identified TsNd in the cuticle, stichocytes and reproductive organs of the parasite. Following immunization, anti-rTsNd IgG levels were increased, and the levels of IgG1 were more significantly higher than that of IgG2a. After a challenge infection with T. spiralis, mice immunized with the rTsNd displayed a 57.7% reduction in adult worms and a 56.9% reduction in muscle larval burden. CONCLUSIONS: TsNd induced a partial protective immunity in mice and could be considered as a novel candidate vaccine antigen against trichinellosis. |
format | Online Article Text |
id | pubmed-4287505 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-42875052015-01-09 Molecular identification of Trichinella spiralis nudix hydrolase and its induced protective immunity against trichinellosis in BALB/c mice Long, Shao Rong Wang, Zhong Quan Liu, Ruo Dan Liu, Li Na Li, Ling Ge Jiang, Peng Zhang, Xi Zhang, Zi Fang Shi, Hai Ning Cui, Jing Parasit Vectors Research BACKGROUND: Nudix hydrolases (Nd) is a widespread superfamily, which is found in all classes of organism, hydrolyse a wide range of organic pyrophosphates and has a ‘housecleaning’ function. The previous study showed that Trichinella spiralis Nd (TsNd) bound to intestinal epithelial cells (IECs), and the vaccination of mice with T7 phage-displayed TsNd polypeptides produced protective immunity. The aim of this study was to clone, express and identify the full-length TsNd and to investigate its immune protection against T. spiralis infection. METHODS: The full-length cDNA sequence of TsNd gene encoding a 46 kDa protein from T. spiralis intestinal infective larvae (IIL) was cloned and identified. The antigenicity of rTsNd was analyzed by Western blot. Transcription and expression of TsNd at T. spiralis different stages were observed by RT-PCR and IFT. The levels of the specific total IgG, IgG1 and IgG2a antibodies to rTsNd were determined by ELISA. The immune protection of rTsNd against T. spiralis infection was investigated. RESULTS: Sequence and phylogenetic analysis revealed that TsNd had a nudix motif located at 226-244aa, which had high homology and the closest evolutionary status with T. pseudospiralis. The rTsNd was obtained after expression and purification. Western blot analysis showed that anti-rTsNd serum recognized the native TsNd protein in crude antigens of muscle larvae (ML), IIL, adult worms (AW) and newborn larvae (NBL), and ES antigens of ML. Transcription and expression of TsNd gene was observed in all developmental stages of T. spiralis (ML, IIL, AW and NBL), with high level expression in IIL. An immunolocalization analysis identified TsNd in the cuticle, stichocytes and reproductive organs of the parasite. Following immunization, anti-rTsNd IgG levels were increased, and the levels of IgG1 were more significantly higher than that of IgG2a. After a challenge infection with T. spiralis, mice immunized with the rTsNd displayed a 57.7% reduction in adult worms and a 56.9% reduction in muscle larval burden. CONCLUSIONS: TsNd induced a partial protective immunity in mice and could be considered as a novel candidate vaccine antigen against trichinellosis. BioMed Central 2014-12-19 /pmc/articles/PMC4287505/ /pubmed/25522912 http://dx.doi.org/10.1186/s13071-014-0600-9 Text en © Long et al.; licensee BioMed Central. 2014 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. 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. |
spellingShingle | Research Long, Shao Rong Wang, Zhong Quan Liu, Ruo Dan Liu, Li Na Li, Ling Ge Jiang, Peng Zhang, Xi Zhang, Zi Fang Shi, Hai Ning Cui, Jing Molecular identification of Trichinella spiralis nudix hydrolase and its induced protective immunity against trichinellosis in BALB/c mice |
title | Molecular identification of Trichinella spiralis nudix hydrolase and its induced protective immunity against trichinellosis in BALB/c mice |
title_full | Molecular identification of Trichinella spiralis nudix hydrolase and its induced protective immunity against trichinellosis in BALB/c mice |
title_fullStr | Molecular identification of Trichinella spiralis nudix hydrolase and its induced protective immunity against trichinellosis in BALB/c mice |
title_full_unstemmed | Molecular identification of Trichinella spiralis nudix hydrolase and its induced protective immunity against trichinellosis in BALB/c mice |
title_short | Molecular identification of Trichinella spiralis nudix hydrolase and its induced protective immunity against trichinellosis in BALB/c mice |
title_sort | molecular identification of trichinella spiralis nudix hydrolase and its induced protective immunity against trichinellosis in balb/c mice |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4287505/ https://www.ncbi.nlm.nih.gov/pubmed/25522912 http://dx.doi.org/10.1186/s13071-014-0600-9 |
work_keys_str_mv | AT longshaorong molecularidentificationoftrichinellaspiralisnudixhydrolaseanditsinducedprotectiveimmunityagainsttrichinellosisinbalbcmice AT wangzhongquan molecularidentificationoftrichinellaspiralisnudixhydrolaseanditsinducedprotectiveimmunityagainsttrichinellosisinbalbcmice AT liuruodan molecularidentificationoftrichinellaspiralisnudixhydrolaseanditsinducedprotectiveimmunityagainsttrichinellosisinbalbcmice AT liulina molecularidentificationoftrichinellaspiralisnudixhydrolaseanditsinducedprotectiveimmunityagainsttrichinellosisinbalbcmice AT lilingge molecularidentificationoftrichinellaspiralisnudixhydrolaseanditsinducedprotectiveimmunityagainsttrichinellosisinbalbcmice AT jiangpeng molecularidentificationoftrichinellaspiralisnudixhydrolaseanditsinducedprotectiveimmunityagainsttrichinellosisinbalbcmice AT zhangxi molecularidentificationoftrichinellaspiralisnudixhydrolaseanditsinducedprotectiveimmunityagainsttrichinellosisinbalbcmice AT zhangzifang molecularidentificationoftrichinellaspiralisnudixhydrolaseanditsinducedprotectiveimmunityagainsttrichinellosisinbalbcmice AT shihaining molecularidentificationoftrichinellaspiralisnudixhydrolaseanditsinducedprotectiveimmunityagainsttrichinellosisinbalbcmice AT cuijing molecularidentificationoftrichinellaspiralisnudixhydrolaseanditsinducedprotectiveimmunityagainsttrichinellosisinbalbcmice |