RNAi-mediated silencing of Trichinella spiralis glutaminase results in reduced muscle larval infectivity

Gao, Yuan; Meng, Xianghe; Yang, Xiao; Meng, Shi; Han, Caixia; Li, Xiaoyun; Wang, Shuang; Li, Wei; Song, Mingxin

doi:10.1186/s13567-021-00921-1

Cited by 7 publications

(5 citation statements)

References 26 publications

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“…The acidic environment in the stomach of the host can also stimulate the effective expression of acid-fast genes of T. spiralis ML. A previous study showed that a glutaminase (TsGLS) plays an important role in the acid-resistant system of T. spiralis ( Gao et al, 2021 ). As shown in Figures 10A,B , when the ML were cultured at pH 2.5 for 18 h, the larval survival was more obviously decreased at pH 2.5 than that at pH 4.5 and 7.4, and the enzyme activity of TsGS was significantly lower than that of the ML at pH 7.4.…”

Section: Discussionmentioning

confidence: 99%

“…Inactive, straight, or C-shaped larvae were assessed as dead larvae. The mobile and wriggly larvae were counted as live ones ( Yang et al, 2020 ; Gao et al, 2021 ). Somatic crude extracts from the ML cultured at various pH values for 18 h were prepared, and TsGS enzymatic activity was assessed as described before ( Wang et al, 2018 ).…”

Section: Methodsmentioning

confidence: 99%

“…AR4 is one of the most important acid-resistant systems in E. coli . Recent studies showed that glutaminase is an important gene in glutamine-dependent acid resistance of T. spiralis ML ( Gao et al, 2021 ). However, there are no reports on the GS role in T. spiralis ML acid resistance.…”

Section: Introductionmentioning

confidence: 99%

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Characterization of a Novel Glutamine Synthetase From Trichinella spiralis and Its Participation in Larval Acid Resistance, Molting, and Development

Zhuo

Wang

Song

et al. 2021

Front. Cell Dev. Biol.

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Trichinella spiralis is a major foodborne parasite worldwide. After the encapsulated muscle larvae (ML) in meat are ingested, the ML are liberated in the stomach of the host and activated into intestinal infectious larvae (IIL), which develop into adult worm after molting four times. A novel glutamine synthetase (TsGS) was identified from T. spiralis IIL at 10 h post-infection, but its biological role in T. spiralis life cycle is not clear. The aim of this study was to investigate the biological characteristics of TsGS and its functions in larval acid resistance, molting, and development. TsGS has a glutamine synthetase (GS) catalytic domain. Complete TsGS sequence was cloned and expressed in Escherichia coli BL21. rTsGS has good immunogenicity. qPCR and Western blotting showed that TsGS was highly expressed at IIL stage, and immunofluorescence revealed that TsGS was principally localized at the cuticle and intrauterine embryos of this nematode. rTsGS has enzymatic activity of natural GS to hydrolyze the substrate (Glu, ATP, and NH4+). Silencing of TsGS gene significantly reduced the IIL survival at pH 2.5, decreased the IIL burden, and impeded larval molting and development. The results demonstrated that TsGS participates in T. spiralis larval acid resistance, molting and development, and it might be a candidate vaccine target against Trichinella molting and development.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Characterization of a Novel Glutamine Synthetase From Trichinella spiralis and Its Participation in Larval Acid Resistance, Molting, and Development

Zhuo

Wang

Song

et al. 2021

Front. Cell Dev. Biol.

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“…Additionally, silencing serine protease, glutathione S-transferase reduced T. spiralis invasion of hosts, affecting worm development and fecundity [21,28,29]. Silencing glutaminase reduced larval infectivity, while silencing glutamine synthetase affected acid tolerance, molting and larval development [30,31]. In this study, three pairs of specific short-fragment siRNAs were designed to silence TsCatL by electroporation, and siRNA-302 was the most effective in silencing TsCatL.…”

Section: Plos Neglected Tropical Diseasesmentioning

confidence: 99%

Trichinella spiralis cathepsin L damages the tight junctions of intestinal epithelial cells and mediates larval invasion

Liu,

Meng,

et al. 2023

PLoS Negl Trop Dis

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Background Cathepsin L, a lysosomal enzyme, participates in diverse physiological processes. Recombinant Trichinella spiralis cathepsin L domains (rTsCatL2) exhibited natural cysteine protease activity and hydrolyzed host immunoglobulin and extracellular matrix proteins in vitro, but its functions in larval invasion are unknown. The aim of this study was to explore its functions in T. spiralis invasion of the host’s intestinal epithelial cells. Methodology/principal findings RNAi significantly suppressed the expression of TsCatL mRNA and protein with TsCatL specific siRNA-302. T. spiralis larval invasion of Caco-2 cells was reduced by 39.87% and 38.36%, respectively, when anti-TsCatL2 serum and siRNA-302 were used. Mice challenged with siRNA-302-treated muscle larvae (ML) exhibited a substantial reduction in intestinal infective larvae, adult worm, and ML burden compared to the PBS group, with reductions of 44.37%, 47.57%, and 57.06%, respectively. The development and fecundity of the females from the mice infected with siRNA-302-treated ML was significantly inhibited. After incubation of rTsCatL2 with Caco-2 cells, immunofluorescence test showed that the rTsCatL2 gradually entered into the cells, altered the localization of cellular tight junction proteins (claudin 1, occludin and zo-1), adhesion junction protein (e-cadherin) and extracellular matrix protein (laminin), and intercellular junctions were lost. Western blot showed a 58.65% reduction in claudin 1 expression in Caco-2 cells treated with rTsCatL2. Co-IP showed that rTsCatL2 interacted with laminin and collagen I but not with claudin 1, e-cadherin, occludin and fibronectin in Caco-2 cells. Moreover, rTsCatL2 disrupted the intestinal epithelial barrier by inducing cellular autophagy. Conclusions rTsCatL2 disrupts the intestinal epithelial barrier and facilitates T. spiralis larval invasion.

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“…In addition, a greater inflammatory response was observed near T. nativa capsules [23]. The literature shows that the immune response of the body depends on numerous factors, such as the type of organism [24], the pH [25], the type of Trichinella (encapsulated/non-encapsulated) [26], and the route of infection (intravenous/oral) [27].…”

Section: Hematological Blood Tests On Trichinella Nativa-infected Rab...mentioning

confidence: 99%

Hematological, biochemical, and serological parameters of experimentally infected rabbits with Trichinella nativa and Trichinella spiralis for early identification of trichinellosis

et al. 2022

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Background and Aim: Trichinellosis remains a dangerous disease for humans and animals, which can lead to a lethal outcome. The study of specific body reactions in response to invasion by different types of Trichinella can help in the early diagnosis of the disease. This study aimed to investigate the hematological, biochemical, and serological characteristics of rabbits experimentally infected with trichinellosis, as well as the possibility of using changes in these parameters at various disease stages for early hematological, biochemical, and serological diagnosis of trichinellosis. Materials and Methods: Three groups of rabbits were orally infected with Trichinella nativa and Trichinella spiralis derived from encysted T. spirtalis larvae in pork muscle samples. The first and second groups were infected with T. nativa and T. spiralis, respectively, while the third group served as control by receiving a physiological solution. An ADVIA 2120i automatic hematology analyzer with a blood smear staining module was used to determine the hematological parameters of rabbits. Antigens were used in an enzyme-linked immunosorbent assay (ELISA) to detect antibodies in the sera of infected rabbits that were supernatants containing excretory-secretory antigens (ES-Ag) and somatic antigen (S-Ag). Results: The detection of biochemical responses to the invasion of T. nativa and T. spiralis isolates was detected and hematological parameters were featured in two cases. Trichinella nativa increased the number of erythrocytes, neutrophils, eosinophils, monocytes, basophils, and thrombocytes on day 7 in rabbits. Creatine kinase (CK) is regarded as the most important indicator for the early detection of parasite invasion. Blood biochemistry showed no active response to T. spiralis infection. However, counts of erythrocytes, neutrophils, lymphocytes, and CK rose significantly. In both color indicators, the number of thrombocytes decreased. Enzyme-linked immunosorbent assay with ES-Ag and S-Ag of these isolates demonstrated the ability to detect antibodies as early as 7 days after infection, with a significant increase in the marker up to 70 days. Conclusion: On the 7th day after infection, blood tests of infected animals revealed CK-N-acetyl-cysteine (18.2%) and neutrophils (43%) when infected with T. nativa and neutrophils (26.7%) and lymphocytes (20%) when infected with T. spiralis. These indicators may serve as specific parameters for the early detection of Trichinella spp. invasion.

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RNAi-mediated silencing of Trichinella spiralis glutaminase results in reduced muscle larval infectivity

Cited by 7 publications

References 26 publications

Characterization of a Novel Glutamine Synthetase From Trichinella spiralis and Its Participation in Larval Acid Resistance, Molting, and Development

Characterization of a Novel Glutamine Synthetase From Trichinella spiralis and Its Participation in Larval Acid Resistance, Molting, and Development

Trichinella spiralis cathepsin L damages the tight junctions of intestinal epithelial cells and mediates larval invasion

Hematological, biochemical, and serological parameters of experimentally infected rabbits with Trichinella nativa and Trichinella spiralis for early identification of trichinellosis

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