Protein change of intestinal epithelial cells induced in vitro by Trichinella spiralis infective larvae

Abstract

The aim of this study was to observe the protein changes of intestinal epithelial cells induced in vitro by Trichinella spiralis infective larvae and their excretory-secretory (ES) or surface antigens and identity the proteins related with invasion. HCT-8 cells were incubated for 2 h in the culture medium contained ES or surface antigens of infective larvae, and observed by Immunofluorescent test (IFT). The infective larvae were inoculated into culture of HCT-8 cells to incubate for 18 h, and the lysates of HCT-8 cells were analyzed by SDS-PAGE and Western blot. IFA showed that normal HCT-8 cells had positively reactions with sera of the infected mice and mice immunized with ES or surface antigens. However, after incubating with ES or surface antigens, HCT-8 cells had stronger positively reaction with the above sera. On Western blot, after cultured with infective larvae, additional seven protein bands (66, 61, 57, 45, 34, 21, and 17 kDa) of HCT-8 cells were recognized by sera of the infected or immunized mice, but three protein bands (48, 43, and 23 kDa) of HCT-8 cells were not recognized by the above sera, compared with normal HCT-8 cells. Our results showed that after cultured with infective larvae the protein components of HCT-8 cell changed, suggesting that additional seven proteins recognized by sera of the infected or immunized mice may be related with invasion of intestinal epithelial cells by infective larvae, these proteins might mediate or facilitate entry into the cells, while the three proteins not recognized by the above sera may be the specific mediators released from the cells which permit invasion.

Comparative proteomic analysis of surface proteins of Trichinella spiralis muscle larvae and intestinal infective larvae

“…Protein samples (20 g) were separated by SDS-PAGE on 5% stacking gels and 12% resolving gels (83 × 73 × 1.0 mm) in a minivertical electrophoresis system (Bio-Rad Laboratories, USA) (Wang et al, 2011). After electrophoresis, the gel was stained with 0.25% Coomassie brilliant blue (CBB) R-250 (Sigma, USA) for 4 h, and then bleached with the eluate.…”

Helminths and intestinal barrier function

“…32,33,[38][39][40][41] Studies in which helminth E/S products are added directly to epithelial monolayers and permeability subsequently assessed are scarce. The E/S products from the nematodes Haemonchus contortus and Teladorsagia circumcincta when applied to the apical surface of monolayers of the human colon-derived Caco2 epithelial cell line lowered the TER by »20% 2 hours post-treatment.…”

Inhibition of mammalian muscle differentiation by excretory secretory products of muscle larvae of Trichinella spiralis in vitro

“…These figures are representative of three independent experiments as growth factors, proteases and cytokine (Guttridge et al 1997;Olwin and Rapraeger 1992;Rao and Kohtz 1995;Zorzano et al 2003). A previous study suggested that ML-ESP of T. spiralis may be combined to the surface of intestinal epithelial cells and induce the specific mediators or signal transduction protein, which could facilitate larvae invasion (Wang et al 2011). From the results presented here, we speculate that factors within the ML-ESP reversibly bound to specific receptors at the surface of the myoblast to modulate cellular proliferation and differentiation via the activation of specific intracellular signalling cascades.…”