Characterization of a Trichinella spiralis 31kDa protein and its potential application for the serodiagnosis of trichinellosis

Cui, Jing; Wang, Li; Sun, Ge; Liu, Li Na; Zhang, Shuai Bing; Liu, Ruo Dan; Zhang, Xi; Jiang, Peng; Wang, Zhong Quan

doi:10.1016/j.actatropica.2014.10.017

Cited by 74 publications

(73 citation statements)

References 37 publications

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“…TspSP-1.2 could play an important role in the larval invasion of host’s intestinal mucosa and might be a potential candidate target for vaccine against T. spiralis infection (Wang B. et al, 2013). The 31 kDa proteins screened from T. spiralis ML ES proteins by immunoproteomics, belongs to the trypsin-like serine protease superfamily, possess significant potential as early diagnostic antigen for trichinellosis (Cui et al, 2015). The TsSP-ZH68 identified in this study was chosen for expression in recombinant protein in E. coli to further evaluate as a reagent for the early serodiagnosis for trichinellosis.…”

Section: Discussionmentioning

confidence: 99%

“…These patients had a history of ingestion of raw or undercooked meat and presented with typical clinical manifestations of trichinellosis (fever related with periorbital or facial oedema, myalgia, and eosinophilia). All these patients had serum specific anti- Trichinella IgG antibody by ELISA with ML ES antigens (Cui et al, 2015; Sun et al, 2015). The optical density values at 490 nm of ten serum samples from patients with early trichinellosis at 19 dpi were 0.54, 0.535, 0.585, 0.615, 0.545, 0.585, 0.56, 0.59, 0.53, and 0.575, respectively; the cut-off value was 0.45.…”

Section: Methodsmentioning

confidence: 99%

“…Trichinella spiralis muscle larvae (ML) excretory-secretory (ES) antigens, recommended by the International Commission on Trichinellosis (ICT), are the most commonly used serodiagnostic antigens for trichinellosis (Gamble et al, 2004), but the specific antibodies against the ML ES antigens are usually negative during early stage of Trichinella infection (Cui et al, 2015), possibly because the majority of ML ES antigens are the stage-specific and not recognized by specific antibodies produced during the intestinal phase (Liu J.Y. et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

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Proteomic Analysis of Trichinella spiralis Adult Worm Excretory-Secretory Proteins Recognized by Sera of Patients with Early Trichinellosis

et al. 2017

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The most commonly used serodiagnostic antigens for trichinellosis are the excretory-secretory (ES) antigens from T. spiralis muscle larvae (ML), but the specific antibodies against the ML ES antigens are usually negative during early stage of Trichinella infection. The recent studies demonstrated that T. spiralis adult worm (AW) antigens were recognized by mouse or swine infection sera on Western blot as early as 7–15 days post-infection (dpi), the AW antigens might contain the early diagnostic markers for trichinellosis. The purpose of this study was to screen early diagnostic antigens in T. spiralis AW ES proteins recognized by sera of early patients with trichinellosis. T. spiralis AW were collected at 72 h post-infection (hpi), and their ES antigens were analyzed by SDS-PAGE and Western blot. Our results showed that 5 protein bands (55, 48–50, 45, 44, and 36 kDa) were recognized by sera of early patients with trichinellosis collected at 19 dpi, and were subjected to shotgun LC–MS/MS and bioinformatics analyses. A total of 185 proteins were identified from T. spiralis protein database, of which 116 (67.2%) proteins had molecular weights of 30∼60 kDa, and 125 (67.6%) proteins with pI 4–7. Bioinformatic analyses showed that the identified proteins have a wide diversity of biological functions (binding of nucleotides, proteins, ions, carbohydrates, and lipids; hydrolase, transferase, and oxidoreductase, etc.). Several enzymes (e.g., adult-specific DNase II, serine protease and serine protease inhibitor) could be the invasion-related proteins and early diagnostic markers for trichinellosis. Moreover, recombinant T. spiralis serine protease (rTsSP-ZH68) was expressed in E. coli and its antigenicity was analyzed by Western blot with the early infection sera. The rTsSP-ZH68 was recognized by sera of infected mice at 8–10 dpi and sera of early patients with trichinellosis at 19 dpi. T. spiralis AW proteins identified in this study, especially serine protease, are the promising early diagnostic antigens and vaccine candidates for trichinellosis.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Proteomic Analysis of Trichinella spiralis Adult Worm Excretory-Secretory Proteins Recognized by Sera of Patients with Early Trichinellosis

et al. 2017

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“…Sometimes the serodiagnosis is negative during the fi rst days of the muscular phase, the combination use of ELISA and IFA tests can make the diagnosis earlier. Other parasitic diseases which manifest fever and eosinophilia in China include acute paragonimiasis, clonorchiasis, schistosomiasis, and sparganosis, a few of patients with trichinellosis might have detectable IgG antibodies cross-reacted with these parasites by ELISA or IFT test that yield false-positive results, which may lead to misinterpretation (Cui et al, 2015). The defi nitive diagnosis of trichinellosis can be made by fi nding larvae in biopsy skeletal muscles (gastrocnemius or deltoid muscles).…”

Section: Discussionmentioning

confidence: 99%

Clinical and etiological study of a small familiar outbreak of trichinellosis in Tibet, China

Wang

Ciren²,

Ren

et al. 2015

Helminthologia

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SummaryThe diagnosis of trichinellosis is rather diffi cult because fever, myalgia and eosinophilia are nonspecifi c, and diagnosis may be delayed. We describe a small familiar outbreak of trichinellosis occurred in early 2009 in Tibet, southwestern China, due to consumption of raw pork. The patients with trichinellosis were diagnosed by muscle biopsy and serological tests (ELISA and IFA) early at 12 days after onset of disease. Both of the pre-encapsulated and encapsulated Trichinella larvae collected from biopsy muscles and residual pork were identifi ed as Trichinella spiralis by multiplex PCR. This is the fi rst species identifi cation of Trichinella isolates from the biopsy muscles of patients with trichinellosis in China.

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“…Serum levels of anti-rTsE antibodies (total IgG, IgG1, IgG2a, and IgA) in all vaccinated mice were tested by ELISA with rTsE [35,36]. Briefly, the ELISA plate was coated using 1 μg/mL rTsE for total IgG and 2 μg/mL rTsE for IgG1, IgG2a and IgA at 37 °C for 2 h. Skimmed milk at a 5% concentration was used for blocking at 37 °C for 1 h. Different dilutions of mouse sera (1:100 for detecting IgG, IgG1 and IgG2a; 1:50 for detecting IgA) were added and incubated at 37 °C for 2 h. Following washes with TBS-0.5% Tween 20 (TBST), 1:10 000 dilutions of HRP-conjugated anti-mouse IgG, IgG1 or IgG2a, and 1:5000 dilutions of HRP-conjugated anti-mouse IgA were added and incubated for 1 h at 37 °C [37].…”

Section: Detection Of Antibody Responses To Rtsementioning

confidence: 99%

Protective immunity in mice vaccinated with a novel elastase-1 significantly decreases Trichinella spiralis fecundity and infection

Zhang

Sun

Bai

et al. 2020

Vet Res

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Trichinella spiralis is an important foodborne parasitic nematode that represents an enormous threat to the food safety of pork meat. The development of a preventive vaccine is valuable for the prevention and control of Trichinella infection in domestic pigs to ensure pork safety. Elastase is a trypsin-like serine protease that hydrolyzes the host's diverse tissue components and participates in parasite penetration, and it might be a novel vaccine target molecule. The aim of this study was to assess the protective immunity produced by vaccination with a novel Trichinella spiralis elastase-1 (TsE) in a mouse model. The results demonstrate that subcutaneous vaccination of mice with rTsE elicited a systemic humoral response (high levels of serum IgG and subclass IgG1/IgG2a and IgA) and significant local enteral mucosal sIgA responses. Anti-rTsE IgG recognized the native TsE at the cuticle, stichosome of intestinal infective larvae and adult worm (AW), and intrauterine embryos of female AW. The rTsE vaccination also produced a systemic and local mixed Th1/Th2 response, as demonstrated by clear elevation levels of Th1 cytokines (IFN-γ, IL-2) and Th2 cytokines (IL-4, IL-10) after spleen, mesenteric lymph node and Peyer's patch cells from immunized mice were stimulated with rTsE. The immunized mice exhibited a 52.19% reduction in enteral AW and a 64.06% reduction in muscle larvae after challenge infection. The immune response triggered by rTsE vaccination protected enteral mucosa from larval intrusion, suppressed larval development and reduced female fecundity. The results indicate that TsE may represent a novel target molecule for anti-T. spiralis vaccines.© The Author(s) 2020. This 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'

show abstract

Characterization of a Trichinella spiralis 31kDa protein and its potential application for the serodiagnosis of trichinellosis

Cited by 74 publications

References 37 publications

Proteomic Analysis of Trichinella spiralis Adult Worm Excretory-Secretory Proteins Recognized by Sera of Patients with Early Trichinellosis

Proteomic Analysis of Trichinella spiralis Adult Worm Excretory-Secretory Proteins Recognized by Sera of Patients with Early Trichinellosis

Clinical and etiological study of a small familiar outbreak of trichinellosis in Tibet, China

Protective immunity in mice vaccinated with a novel elastase-1 significantly decreases Trichinella spiralis fecundity and infection

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