Abstract
Simple SummaryHaemonchus contortus excretory/secretory products (HcESPs) contain many proteins that can perform various functions including modulating the host immune response. Recent studies indicate that IL-9 can be secreted by a specialized population of T cells called Th9 cells, which mediate anti-parasite immunity. Furthermore, HcESPs could enhance goat peripheral blood mononuclear cells (PBMCs) derived Th9 cells production. Ephrin domain containing protein (EPH) was identified as one of the HcESPs that can be isolated from different stages of this helminth. Nonetheless, the understanding of immunomodulatory roles of EPH on Th9 and other immune cells remains limited. In this study, the correlation between recombinant H. contortus Ephrin domain containing protein(rHcEPH)and goat PBMCs significantly enhanced Th9 cells differentiation, IL-9 expression, cell apoptosis efficiency, and cell migration, whereas cell proliferation was suppressed significantly depending on the concentration. Our findings illustrated that rHcEPH protein is linked to modulate the host immune cells and could enhance protective immunity by inducing Th9 cells secreted IL-9 in vitro.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.
Highlights
Haemonchosis is a parasitic disease caused by gastrointestinal nematode Haemonchus contortus [1].Goats and sheep can become infected upon ingestion of infective larvae which may result in anemia, diarrhea, decrease in weight, and death in some cases [2,3]
Our study indicated that rHcEPH as a constituent of HcESPs might interfere with modulatory activity on peripheral blood mononuclear cells (PBMCs) proliferation and cell death regulation; the actual mechanism and pathways involved in the adjustment of this apoptosis and proliferation need to be studied further
It was highlighted that, as an important and active constituent of HcESPs localized in adult worms, rHcEPH plays a crucial role in the cellular immune regulations
Summary
Haemonchosis is a parasitic disease caused by gastrointestinal nematode Haemonchus contortus [1].Goats and sheep can become infected upon ingestion of infective larvae which may result in anemia, diarrhea, decrease in weight, and death in some cases [2,3]. Haemonchosis is a parasitic disease caused by gastrointestinal nematode Haemonchus contortus [1]. H. contortus, mediated by parasite-derived effector molecules, is believed to play important roles in the establishment of long-lasting infection in the host. Several attempts were made for the recognition of immunogenic molecules derived from ESPs with potential for vaccine use, such as transthyretin domain- containing protein [1], 14-3-3 Isoform 2 [4], and glyceraldehyde-3-phosphate dehydrogenase [5]. It was demonstrated previously that Ephrin domain-containing protein (EPH) from HcESPs from this parasite in various developmental stages [8]. Ephrin plays important roles during anti-cancer therapies, hematopoiesis and proliferation of cells [9,10,11]. The exact biological function and immunological characters of HcEPH are not clear
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