Trichinella spiralis: RNAi-mediated silencing of serine protease results in reduction of intrusion, development and fecundity
https://doi.org/10.47665/tb.37.4.932
- Author:
Yang, D.Q.
1
;
Zeng, J.
1
;
Sun, X.Y.
1
;
Yue, X.
1
;
Hu, C.X.
1
;
Jiang, P.
1
;
Liu, R.D.
1
;
Cui, J.
1
;
Wang, Z.Q.
1
Author Information
1. Department of Parasitology, Medical College, Zhengzhou University, 40 Daxue Road, Zhengzhou 450052, Peoples Republic of China
- Publication Type:Journal Article
- From:Tropical Biomedicine
2020;37(No.4):932-946
- CountryMalaysia
- Language:English
-
Abstract:
In previous studies, a Trichinella spiralis serine protease (TsSP) was identified in
excretion/secretion (ES) products from intestinal infective L1 larvae (IIL1) using
immunoproteomics. The complete cDNA sequence of TsSP gene was 1372 bp, which encoded
429 amino acids with 47.55 kDa. The TsSP was transcribed and expressed at all T. spiralis life
cycle phases, as well as mainly located at the cuticle and stichosome of the parasitic nematode.
Recombinant TsSP bind to intestinal epithelial cells (IEC) and promoted larva invasion, however,
its exact function in invasion, development and reproduction are still unknown. The aim of
this study was to confirm the biological function of TsSP during T. spiralis invasion and
growth using RNA interference (RNAi) technology. The results showed that on 1 day after
electroporation using 2.5 µM siRNA156, TsSP mRNA and protein expression of muscle larvae
(ML) was suppressed by 48.35 and 59.98%, respectively. Meanwhile, silencing of TsSP gene
by RNAi resulted in a 61.38% decrease of serine protease activity of ML ES proteins, and a
significant reduction of the in vitro and in vivo invasive capacity of IIL1 to intrude into the
IEC monolayer and intestinal mucosa. When mice were infected with siRNA 156-transfected
larvae, adult worm and muscle larva burdens were decreased by 58.85 and 60.48%,
respectively. Moreover, intestinal worm growth and female fecundity were evidently inhibited
after TsSP gene was knockdown, it was demonstrated that intestinal adults became smaller
and the in vitro newborn larval yield of females obviously declined compared with the
control siRNA group. The results indicated that knockdown of TsSP gene by RNAi significantly
reduced the TsSP expression and enzymatic activity, impaired larvae intrusion and growth,
and lowered the female reproductive capacity, further verified that TsSP might participate in
diverse processes of T. spiralis life cycle, it will be a new prospective candidate molecular
target of anti-Trichinella vaccines.
- Full text:8.2020my1179.pdf
