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

Abstract

Excretory/secretory proteins of Haemonchus contortus (HcESPs) intermingle comprehensively with host immune cells and modulate host immune responses. In this study, H contortus ES antigen named as elongation factor 1 alpha (HcEF‐1α) was cloned and expressed. The influences of recombinant HcEF‐1α on multiple functions of goat peripheral blood mononuclear cells (PBMCs) were observed in vitro. Immunoblot analysis revealed that rHcEF‐1α was recognized by the serum of goat infected with H contortus. Immunofluorescence analysis indicated that rHcEF‐1α was bound on surface of PBMCs. Moreover, the productions of IL‐4, TGF‐β1, IFN‐γ and IL‐17 of cells were significantly modulated by the incubation with rHcEF‐1α. The production of interleukin IL‐10 was decreased. Cell migration, cell proliferation and cell apoptosis were significantly increased; however, nitric oxide production (NO) was significantly decreased. The MHC II molecule expression of cells incubated with rHcEF‐1α was increased significantly, whereas MHC‐I was not changed as compared to the control groups (PBS control and pET32a). These findings indicated that rHcEF‐1α protein might play essential roles in functional regulations of HcESPs on goat PBMC and mediate the immune responses of the host during host‐parasite relationship.

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

“…Previous studies have shown that various HcESP-derived protein molecules, such as rHcMTF-12 [ 54 ], rHCRD [ 37 ], rHc-GDC [ 40 ], rHcEF-1α [ 46 ], rHcES-15 [ 34 ], rMiro-1 [ 55 ], HCcyst-3 [ 56 ] and RHC-AK [ 57 ], can bind to PBMCs in vitro and regulate cellular immune function. Our studies showed that rHcGOB can bind to goat PBMCs and activate the IL-10/TGF-β/STAT3 pathway in PBMCs in vitro.…”

“…The transcriptional levels of mRNA were detected by qPCR, and the β-actin gene was used as the reference gene. In accordance with a previous study [ 46 ], the primer sequences shown in Additional file 1 : Table S2 were used. The calculation was carried out according to the comparative Ct (2 −ΔΔCt ) method.…”

Histidine acid phosphatase domain-containing protein from Haemonchus contortus is a stimulatory antigen for the Th1 immune response of goat PBMCs

“…The reaction system contained 5 μl 2× ChamQ SYBR qPCR Master Mix (Vazyme Biotech Co., Ltd.), 3.6 μl ddH 2 O, 0.2 μl forward and reverse primers and 1 μl cDNA. Primer sequences are shown in Additional file 1 : Table S2 [ 23 , 24 ]. To calculate raw cycle thresholds (Ct), the relative Ct (2 −ΔΔCt ) method was used with ABI Prism 7500 software (Applied Biosystems).…”

Fasciola gigantica tegumental calcium-binding EF-hand protein 4 exerts immunomodulatory effects on goat monocytes

“…The monocytes were separated from freshly collected PBMCs, and washed with PBS (Ca 2+ /Mg 2+ -free, pH 7.4). Cells numbers were adjusted to 1 × 10 5 cells/ml in RPMI-1640 supplemented with 10% FBS, 100 U/ml penicillin and 100 mg/ml streptomycin (Gibco Life Technologies); then incubated with rFg-CaBP4, or control buffer, at 37 °C in 5% CO 2 for 2 h. Protein binding was examined using an immunofluorescence assay [ 34 ]. The monocytes were washed and added to poly- L -lysine-treated slides, fixed with 4% paraformaldehyde for 30 min and permeabilized with 1% Triton X-100/PBS.…”