Unveiling the Immunomodulatory Characteristics of Haemonchus contortus Ephrin Domain Containing Protein in the Parasite–Host Interactions

Abstract

Ephrin domain containing protein (EPH), a significant excreted and secreted product (ESPs) of Haemonchus contortus, has been identified to have antigenic functions. Over the past years, a new subset of CD4 + T named as T helper 9 cells that secrete interleukin-9 (IL-9) as a signature cytokine is associated with tumor immunity and allergy. Nonetheless, the understanding of immunomodulatory roles of EPH on goat Th9 and other immune cells remains limited. Herein, EPH from H. contortus (HcEPH) was cloned and expressed in pET-28a. Immunofluorescence assay (IFA) was carried-out to localize rHcEPH within H. contortus adult worms and to bind with goat peripheral blood mononuclear cells (PBMCs). Besides, the impact of rHcEPH on signature cytokine IL-9 expression in goat PBMCs was evaluated. Flow cytometry was employed to examine Th9 cells production and cell apoptosis. The results revealed success in the expression and localization of rHcEPH in surface of adult H. contortus gut sections. According to IFA analysis, the rHcEPH protein was capable to react precisely with anti-H. contortus antibodies. Further functional analysis showed that correlation between rHcEPH and host PBMCs significantly enhanced Th9 cell differentiation, IL-9 expression, cell apoptosis efficiency, and cell migration, whereas cell proliferation was suppressed significantly depending on the concentration. Our observations indicated that rHcEPH protein is linked to modulate the host immune cells and could enhance protective immunity by inducing Th9 responses.

The GT1-TPS Structural Domain Protein From Haemonchus contortus Could Be Suppressive Antigen of Goat PBMCs

“…The rHc-TPS protein (30 µg) was separated by 12% SDS-PAGE and transferred to polyvinylidene difluoride (PVDF) membranes (Millipore, USA) as described previously ( 30 ). Afterward, the non-specific binding site by skim milk (5%) in Tris-buffered saline solution consisting of 0.1% Tween-20 solution (TBST) was blocked.…”

In vitro characterization of Haemonchus contortus trehalose-6-phosphate phosphatase and its immunomodulatory effects on peripheral blood mononuclear cells (PBMCs)

“…Adult H. contortus worms were obtained as previously described [ 33 ]. Total RNA was extracted from adult worms by the TRIzol reagent (Invitrogen, Carlsbad, CA, USA) method [ 34 ].…”

“…The binding ability of rHcGOB protein to goat peripheral blood mononuclear cells was detected by IFA as previously described [ 33 , 42 , 43 ], . The fresh goat PBMCs were inoculated with 10 μg/ml rHcGOB or PBS in a 24-well plate (1 ml/well) and the plate incubated at 37 °C with 5% CO 2 for 2 h. The cells were then washed with PBS, placed on poly l -lysine-coated slides for 20 min and fixed with 4% phosphate-buffered paraformaldehyde at room temperature.…”

Nanoparticles of Chitosan/Poly(D,L-Lactide-Co-Glycolide) Enhanced the Immune Responses of Haemonchus contortus HCA59 Antigen in Model Mice

“…[5][6][7] H. contortus excretory and secretory products (HcESPs), which comprised numerous proteins, 8 could execute different functions, such as host protein degradation, tissue penetration, and the induction of Th9 immune responses. 5,9 HCA59 protein belonged to telomere length and silencing protein 1 (TLS1) family and was related to the spliceosome. It was overexpressed in multiple cancer cell lines in human.…”