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In vitro silencing of a serine protease inhibitor suppresses Trichinella spiralis invasion, development, and fecundity

  • Helminthology - Original Paper
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Abstract

In a previous study, immunoproteomics was used to identify a serine protease inhibitor (TsSPI) of T. spiralis excretory/secretory (ES) proteins that exhibited an inhibitory effect on trypsin enzymatic activity, but the precise role of TsSPI on worm infection and development in its host is not well understood. The objective of the present study was to use RNA interference to ascertain the function of TsSPI in larval invasion and growth. TsSPI-specific small interference RNAs (siRNAs) were delivered to muscle larvae (ML) to silence TsSPI expression by electroporation. Four days after electroporation, the ML transfected with 2 μM siRNA-653 exhibited a 75.75% decrease in TsSPI transcription and a 69.23% decrease in TsSPI expression compared with control ML. Although the silencing of TsSPI expression did not decrease worm viability, it significantly suppressed the larval invasion of intestinal epithelium cells (IEC) (P < 0.01), and the suppression was siRNA dose-dependent (r = 0.981). The infection of mice with siRNA-653-treated ML produced a 63.71% reduction of adult worms and a 72.38% reduction of muscle larvae. In addition, the length of the adults, newborn larvae, and ML and the fecundity of female T. spiralis from mice infected with siRNA-treated ML were obviously reduced relative to those in the control siRNA or PBS groups. These results indicated that the silencing of TsSPI by RNAi suppressed larval invasion and development and decreased female fecundity, further confirming that TsSPI plays a crucial role during the T. spiralis lifecycle and is a promising molecular target for anti-Trichinella vaccines.

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Funding

This work was supported by the National Natural Science Foundation of China (No. 81572024, U1704284).

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Authors and Affiliations

  1. Department of Parasitology, Medical College, Zhengzhou University, 40 Daxue Road, Zhengzhou, 450052, People’s Republic of China

    Fan Yang, Da Qi Yang, Yan Yan Song, Kai Xia Guo, Ya Lan Li, Shao Rong Long, Peng Jiang, Jing Cui & Zhong Quan Wang

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  1. Fan Yang
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  2. Da Qi Yang
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  7. Peng Jiang
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Correspondence to Peng Jiang, Jing Cui or Zhong Quan Wang.

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All animal experimental procedure was approved by the Life Science Ethics Committee of Zhengzhou University (No. SCXK 2015–0005).

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Yang, F., Yang, D.Q., Song, Y.Y. et al. In vitro silencing of a serine protease inhibitor suppresses Trichinella spiralis invasion, development, and fecundity. Parasitol Res 118, 2247–2255 (2019). https://doi.org/10.1007/s00436-019-06344-4

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