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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...

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Autores principales: 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
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
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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.
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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
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