Molecular identification and characterization of Trichinella spiralis proteasome subunit beta type-7

Particle and Fibre Toxicology

Jing Cui¹  Zhong Quan Wang¹  Hui Jun Ren²  Xi Zhang¹  Peng Jiang¹  Shuai Bing Zhang¹  Chun Ying Liu¹  Ge Ge Sun¹  Ruo Dan Liu¹  Ling Ge Li¹  Wei Yang¹ 

[1] Department of Parasitology, Medical College of Zhengzhou University, Zhengzhou 450052, P. R. China;Department of Laboratorial Medicine, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, P. R. China

关键词: Immune protection;    Invasion;    Proteasome subunit beta type-7 (Tspst);    Trichinella spiralis;   

Others  :  1147910
DOI  :  10.1186/s13071-014-0626-z

 received in 2014-08-23, accepted in 2014-12-21,  发布年份 2015

【 摘 要 】

Background

Previous study showed that Trichinella spiralis proteasome subunit beta type-7 (Tspst) gene is an up-regulated gene in intestinal infective larvae (IIL) compared to muscle larvae (ML), which was screened by using suppression subtractive hybridization (SSH) and confirmed by real-time PCR. Tspst may be related to the larval invasion of intestinal epithelial cells (IECs). The aim of this study was to identify Tspst and to investigate its immune protection against intestinal T. spiralis infection.

Methods

The Tspst gene encoding a 29 kDa protein from T. spiralis infective larvae was cloned, and recombinant Tspst protein (rTspst) was produced in an Escherichia coli expression system. The rTspst was used to immunize BALB/c mice. Anti-rTspst antibodies were used to determine the immunolocolization of Tspst in the parasite. Transcription and expression of Tspst at T. spiralis different developmental stages were observed by RT-PCR and immunofluorescence test (IFT). The in vitro or in vivo immune protection of anti-rTspst serum or rTspst against intestinal T. spiralis infection in BALB/c mice was evaluated.

Results

Anti-rTspst serum recognized the native Tspst protein with 29 kDa in ML crude antigens. Transcription and expression of gene was observed at all T. spiralis different developmental stages (IIL, adult worms, newborn larvae, and ML). An immunolocalization analysis identified Tspst in the cuticle and internal organs of the parasite. An in vitro invasion assay showed that, when anti-rTspst serum, serum of mice infected with T. spiralis or normal mouse serum were added to the medium, the invasion rate of the infective larvae in an IEC monolayer was 25.2%, 11.4%, and 79%, respectively (P < 0.05), indicating that anti-rTspst serum partially prevented the larval invasion of IECs. After a challenge infection with T. spiralis muscle larvae, mice immunized with rTspst conferred a 45.7% reduction in adult worm burden in intestines.

Conclusions

In the present study, Tspst was first identified and characterized. Tspst is an invasion-related protein of T. spiralis IIL and could be considered as a potential vaccine candidate antigen against intestinal T. spiralis infection that merits further study.

【 授权许可】

   
2015 Yang et al.; licensee BioMed Central.

【 图 表 】

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