The immunogenic maturation of goat monocyte-derived dendritic cells and upregulation of toll-like receptors by five antigens of Haemonchus contortus in-vitro

• Published in: Research in veterinary science Vol. 136; pp. 247 - 258
• Main Authors: Hasan, Muhammad Waqqas, Haseeb, Muhammad, Ehsan, Muhammad, Gadahi, Javaid Ali, Wang, Qiangqiang, Memon, Muhammad Ali, Aleem, Muhammad Tahir, Lakho, Shakeel Ahmed, Yan, Ruo Feng, Xu, Li Xin, Song, Xiao Kai, Li, Xiangrui
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01-05-2021; Elsevier Limited
• Subjects: Activation of the immune response; Adenosine diphosphate; ADP-ribosylation; ADP-ribosylation factor 1; Animals; Antigens; Arginine kinase; CD80 antigen; Cell culture; Chromatography; Cytokines; Dendritic cell functions; Dendritic cells; Drug resistance; E coli; Elongation; Enzyme-linked immunosorbent assay; Flow cytometry; Gene expression; Goat monocytes; Haemonchus contortus; Haemonchus contortus, Immuno-stimulatory antigens; Immune system; Immunofluorescence; Immunogenicity; Immunomodulation; Infections; Interleukin 10; Interleukin 12; Kinases; Leukocytes (mononuclear); Major histocompatibility complex; Mammals; Maturation; Monocytes; Parasites; Pathogens; Peripheral blood mononuclear cells; Plasmids; Polymerase chain reaction; Proteins; Receptors; Ribosylation; Signal transduction; Stimulation; Toll-like receptors; Tumor necrosis factor-α; Up-regulation; Veterinary medicine; Wnt protein; γ-Interferon; United States > US; ARFs; Haemonchus contortus, Immuno-stimulatory antigens; RT; HcEF1-α; rHcEF1-α; IFA; H; md-DCs; PFA; HcES-15; Activation of the immune response; LPS; Dendritic cell functions; BSA; IFN-γ; DCs; GM-CSF; RT-PCR; rHcARF1; HcESPs; rHcAK; rHcES-15; PBS; HcES-24; E/S protein; HcAK; IL-4; DAPI; AK; IL-12; PBMC; IL-10; TNF-α; rHcES-24; Goat monocytes
• ISSN: 0034-5288; 1532-2661
• DOI: 10.1016/j.rvsc.2021.03.007

Previously, it was found that several proteins of Haemonchus contortus were involved in the stimulation of the host immune system. However, the information about the selection of superlative antigens with immunogenic efficacies on host DCs is lacking. In the current study, the stimulatory effects of five recombinant proteins (elongation factor-1α, arginine kinase, ES-15, ES-24, and ADP-ribosylation factor 1) of H. contortus on the maturation of goat monocyte-derived dendritic cells (md-DCs) were reported. Recombinant proteins were purified separately in E. coli expression and incubated with isolated goat peripheral blood mononuclear cells (PBMC). Immunofluorescence assay (IFA) results confirmed the binding of these molecules to the md-DC's surface as compared to control groups. In the flow cytometry analysis, recombinant proteins induced md-DC stimulation via the up-regulation of the expression of the costimulatory molecule (CD80) and MHC-II. Quantitative RT-PCR data showed a significant increase in the expression of specific genes of the WNT and toll-like receptor (TLR) signaling pathways. The result of ELISA indicated the higher levels of cytokine (IL-10, IL-12, IFN-γ, and TNF-α) secretion in the md-DC compared to the negative (pET-32a His-Tag) and blank (PBS) control groups. The data gives valuable support in the selection of potential antigens for future studies on the immunomodulation of the host against the infection of H. contortus.
•H. contortus recombinant antigens induced goat md-DC's maturation and differentiation.•CD-80 and MHC-II molecules were upregulated after incubation of H. contortus recombinant antigens.•The gene expression of Toll-like receptors was upregulated in response to H. contortus recombinant antigens and mediated the stimulation of md-DC against the infection.•The recombinant antigens were involved in modulating the host immune response and might contribute to inhibit H. contortus invasion.