Proteomic Analysis of the Excretory and Secretory Proteins of Haemonchus contortus (HcESP) Binding to Goat PBMCs In Vivo Revealed Stage-Specific Binding Profiles

Abstract

Haemonchus contortus is a parasitic gastrointestinal nematode, and its excretory and secretory products (HcESPs) interact extensively with the host cells. In this study, we report the interaction of proteins from HcESPs at different developmental stages to goat peripheral blood mononuclear cells (PBMCs) in vivo using liquid chromatography-tandem mass spectrometry. A total of 407 HcESPs that interacted with goat PBMCs at different time points were identified from a H. contortus protein database using SEQUEST searches. The L4 and L5 stages of H. contortus represented a higher proportion of the identified proteins compared with the early and late adult stages. Both stage-specific interacting proteins and proteins that were common to multiple stages were identified. Forty-seven interacting proteins were shared among all stages. The gene ontology (GO) distributions of the identified goat PBMC-interacting proteins were nearly identical among all developmental stages, with high representation of binding and catalytic activity. Cellular, metabolic and single-organism processes were also annotated as major biological processes, but interestingly, more proteins were annotated as localization processes at the L5 stage than at the L4 and adult stages. Based on the clustering of homologous proteins, we improved the functional annotations of un-annotated proteins identified at different developmental stages. Some unnamed H. contortus ATP-binding cassette proteins, including ADP-ribosylation factor and P-glycoprotein-9, were identified by STRING protein clustering analysis.

Tropomyosin: An Excretory/Secretory Protein from Haemonchus contortus Mediates the Immuno-Suppressive Potential of Goat Peripheral Blood Mononuclear Cells In Vitro

“…In our previous research, tropomyosin was identified as an excretory and secretory protein (ESP) that binds to goat PBMCs at the L4 and L5 developmental stages of H. contortus [20], and we also demonstrated that HcESPs exhibit suppressive regulatory roles in their interaction with PBMCs [21]. In our recent study, recombinant tropomyosin was found to have high immunodiagnostic potential for early stage diagnosis of H. contortus infection in goats [22].…”

Recombinant cold shock domain containing protein is a potential antigen to detect specific antibody during early and late infections of Haemonchus contortus in goat

“…Antigenicity, sensitivity and specificity may be influenced by the secretion time of antigen. It has been reported earlier that rHc-CS releases at 14 and 60 D.P.I and antibodies against this molecule are produced [23]. Another possibe reason might be the type of antigen used in study.…”

“…Furthermore, recombinant protein, having high immunoreactive concentration, may act as best immunodiagnostic antigen by achieving high sensitivity and specificity [29][30][31][32][33][34][35][36]. Recombinant Hc-CS is a major constituent of HcES proteins [23] which may have potential to diagnose H. contortus at early stage of infection. This study was designed to evaluate the specific antibodies detection during early and late infections of H. contortus using immunoblotting and indirect-ELISA based on rHc-CS in experimentally infected goat.…”

“…In our previous proteomic study, interaction of H. contortus (Hc)ESPs with host peripheral blood cells at different developmental stages was reported. The Cold Shock domain containing protein (CS) is one of these HcESPs, that binds to goat PBMCs at L 4 and L 5 H. contortus development stages [23]. Hence the presence of CS protein may serve for diagnostic purposes as biomarker [24].…”

Recombinant elongation factor 1 alpha of Haemonchus contortus affects the functions of goat PBMCs

“…Previously, in the proteomic analysis of ESPs from H contortus , EF‐1α protein was recognized as binding molecule to goat PBMCs at L4, L5 and early adult developmental stages of H contortus in vivo 23 . Meanwhile, the immuno‐suppressant role of HcESPs on goat PBMCs in vitro was also demonstrated 24 .…”