Identification of a novel methyltransferase-type 12 protein from Haemonchus contortus and its effects on functions of goat PBMCs

• Published in: Parasites & vectors Vol. 13; no. 1; p. 154
• Main Authors: Ehsan, Muhammad, Gadahi, Javaid A, Liu, Tingqi, Lu, Mingmin, Wang, Yujian, Hasan, Muhammad W, Haseeb, Muhammad, Yan, Ruofeng, Xu, Lixin, Song, Xiaokai, Zhu, Xing-Quan, Li, Xiangrui
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 30.03.2020; BioMed Central; BMC
• Subjects: Animal cuticle; Animals; Antibodies; Bacteria; Barber's pole worm; Biological activity; Biotechnology; Bone morphogenetic proteins; Cell proliferation; Cellular signal transduction; Cloning; Cytokines; Damage localization; Deoxyribonucleic acid; DNA; DNA damage; DNA methylation; DNA polymerases; DNA repair; E coli; Enzymes; Escherichia coli; Experiments; Fluorescent antibody technique; Fusion protein; Gene regulation; Genes; Genetic aspects; Goat; Goats; Haemonchus contortus; Health aspects; Host-parasite interactions; Host-parasite relationships; Immune response regulation; Immunofluorescence; Infections; Interleukin 10; Interleukin 2; Interleukin 4; Interleukin 6; Intestine; Localization; Methyltransferase; Methyltransferase-type 12; Nitric oxide; Nitrogen oxides; Nucleic acids; Nucleotide sequence; Parasites; PBMC; PCR; Pets; Pharynx; Polymerase chain reaction; Proliferation; Proteins; Recombinants; Ribonucleic acid; RNA; RNA-directed DNA polymerase; Secretion; Signal processing; Signal transduction; Therapeutic applications; Transcription; Transcription (Genetics); Transferases; Transforming growth factor-b1; Transforming growth factors; γ-Interferon; China; Beijing China; United States > US; Haemonchus contortus; Goat; Methyltransferase-type 12; PBMC; Host–parasite interactions
• ISSN: 1756-3305; 1756-3305
• DOI: 10.1186/s13071-020-04028-y

Methyltransferases (MTFs) are broad range of enzymes, which are ubiquitously expressed in diverse organisms ranging from bacteria to animals. MTFs proteins have been associated with various biological/cellular processes including transcriptional regulation, subcellular protein and RNA localization, signal transduction and DNA-damage repair. However, the role of MTFs in immune mechanism during host-parasite interaction has not been addressed yet. An open reading frame (764 bp) of methyltransferase-type 12 gene of H. contortus denoted as HcMTF-12, was successfully cloned using reverse transcriptase-polymerase chain reaction (RT-PCR) followed by prokaryotic expression in Escherichia coli BL21 (DE3 strain). The recombinant HcMTF-12 protein (rHcMTF-12) was about 47 kDa along with a fusion vector protein of 18 kDa. Immunoblot results identified the native protein MTF-12 with antibodies produced in rats against rHcMT-12, whereas rHcMTF-12 protein was recognized with sera of goat experimentally infected with H. contortus. Immunohistochemical analysis revealed that the native MTF-12 protein was mainly located in the periphery (cuticle) of parasite sections as well as within the pharynx and intestinal region. An immunofluorescence assay validated that rHcMTF-12 attached to the surface of goat PBMCs. Furthermore, the cytokines transcription of IL-2, IFN-γ and IL-4 transcripts of PBMCs incubated with rHcMTF-12 were enhanced in a dose-dependent manner. The secretion of TGF-β1 and IL-10 was significantly decreased. However, IL-6 production was not significantly different as compared to the control groups. Moreover, the migration activity and nitric oxide (NO) production by PBMCs were induced considerably, whereas the proliferation of PBMCs cells was negatively affected when incubated with the rHcMTF-12 protein. Our findings suggest that HcMTF-12 significantly mediated the functions of PBMCs, and it might be a potential candidate for therapeutic interventions against haemonchosis.