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

Muhammad Waqqas Hasan; Muhammad Haseeb; Muhammad Ehsan; Javaid Ali Gadahi; Qiangqiang Wang; Muhammad Ali Memon; Muhammad Tahir Aleem; Shakeel Ahmed Lakho; Ruo Feng Yan; Li Xin Xu; Xiao Kai Song; Xiangrui Li

doi:10.1016/j.rvsc.2021.03.007

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

Res Vet Sci. 2021 May:136:247-258. doi: 10.1016/j.rvsc.2021.03.007. Epub 2021 Mar 9.

Authors

Affiliations

¹ MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu 210095, PR China. Electronic address: 2015207037@njau.edu.cn.
² MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu 210095, PR China. Electronic address: 2016207041@njau.edu.cn.
³ MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu 210095, PR China; State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu, China. Electronic address: mehsan124@gmail.com.
⁴ MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu 210095, PR China; Department of Veterinary Parasitology, Sindh Agriculture University Tandojam, Pakistan.
⁵ MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu 210095, PR China. Electronic address: 2016107053@njau.edu.cn.
⁶ MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu 210095, PR China. Electronic address: 2016207040@njau.edu.cn.
⁷ MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu 210095, PR China. Electronic address: 2018207076@njau.edu.cn.
⁸ MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu 210095, PR China. Electronic address: 2017207046@njau.edu.cn.
⁹ MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu 210095, PR China. Electronic address: yanruofeng@njau.edu.cn.
¹⁰ MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu 210095, PR China. Electronic address: xulixin@njau.edu.cn.
¹¹ MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu 210095, PR China. Electronic address: songxiaokai@njau.edu.cn.
¹² MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu 210095, PR China. Electronic address: lixiangrui@njau.edu.cn.

PMID: 33721712
DOI: 10.1016/j.rvsc.2021.03.007

Abstract

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.

Keywords: Activation of the immune response; Dendritic cell functions; Goat monocytes; Haemonchus contortus, Immuno-stimulatory antigens.

MeSH terms

Animals
Antigens, Helminth / immunology*
Dendritic Cells / immunology*
Goats / immunology*
Haemonchus / immunology*
Monocytes / immunology
Toll-Like Receptors / genetics*
Toll-Like Receptors / metabolism
Up-Regulation*

Substances

Antigens, Helminth
Toll-Like Receptors