Identification of excretory and secretory proteins from Haemonchus contortus inducing a Th9 immune response in goats

Abstract

Th9 cells have been shown to play crucial roles in anti-parasite immunity, pathogenic microbe infection, and allergy. Previous studies have demonstrated that Haemonchus contortus excretory and secretory proteins (HcESPs) induce the proliferation of Th9 cells and alter the transcriptional level of IL-9 as well as its related pathways in the Th9 immune response after infection. However, the exact molecule(s) in HcESPs inducing the Th9 immune response is not yet known. In this study, flow cytometry, co-immunoprecipitation (Co-IP) and shotgun liquid chromatography tandem-mass spectrometry (LC–MS/MS) were used, and a total of 218 proteins from HcESPs that might interact with goat Th9 cells were identified. By in vitro culture of Th9 cells with HcESPs, 40 binding proteins were identified. In vivo, 38, 47, 42 and 142 binding proteins were identified at 7, 15, 35 and 50 days post-infection (dpi), respectively. Furthermore, 2 of the 218 HcESPs, named DNA/RNA helicase domain containing protein (HcDR) and GATA transcription factor (HcGATA), were confirmed to induce the proliferation of Th9 cells and promote the expression of IL-9 when incubated with goat peripheral blood mononuclear cells (PBMCs). This study represents a proteomics-guided investigation of the interactions between Th9 cells and HcESPs. It provides a new way to explore immunostimulatory antigens among HcESPs and identifies candidates for immune-mediated prevention of H. contortus infection.

Haemonchus contortus HcL6 promoted the Th9 immune response in goat PBMCs by activating the STAT6/PU.1/NF-κB pathway

“…Total Th9 cells in goat PBMCs were sorted by the flow-sorting cytometry method as described elsewhere [ 15 ]. Briefly, PBMCs were cultured in RPMI-1640 media containing rHcL6 for 2 h at 37 °C in a humidified 5% CO 2 incubator.…”

“…In a previous study, 218 H. contortus excretory-secretory proteins (HcESPs) interacting with goat Th9 cells were identified by liquid chromatography-tandem mass spectrometry (LC‒MS/MS) analysis. H. contortus ribosomal protein L6 domain DE-containing protein (HcL6) was detected among the proteins interacting with Th9 cells [ 15 ]. Research should be conducted to identify the molecules that directly or indirectly regulate Th9 cells at the parasite-host interface.…”

Epidemiology and phylogeny of Haemonchus contortus through internal transcribed spacer 2 gene in small ruminants

“…In the near future, controlling some parasites with current anthelminthic medications like oxfendazole, levamisole, and ivermectin may become more challenging ( 10 ). H. contortus has evolved different strategies to evade host immune response during infection ( 12 , 13 ). Parasitic nematodes cause a long-term infection in the host, generally with just a modest inflammatory response.…”

Comparative proteomics analysis of adult Haemonchus contortus isolates from Ovis ammon

“…The proteomic analysis of glycoproteins of H. contortus showed that 43 proteins were identified in the adult stage, and 34 and 35 proteins were bound by Lectin galacto side-binding soluble (LGALS)-11 and LGALS-14, respectively ( Sakthivel et al., 2018 ). Shotgun LC−MS/MS was used to identify 218 H. contortu s excretory and secretory proteins Haemonchus contortus Excrete Secretion Proteins (HcESPs) that interact with goat Th9 cells, which provides a new way to explore immunostimulatory antigens among HcESPs of H. contortus infection ( Liang et al., 2022 ). Subsequently, iTRAQ-coupled LC−MS/MS was also applied to analyze the differentially expressed proteins (DEPs) in adult parasites between water buffalo and yellow cattle, and 131 DEPs were identified, including 46 upregulated proteins and 85 downregulated proteins ( He et al., 2020 ).…”