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

• Published in: Parasites & vectors Vol. 14; no. 1; p. 611
• Main Authors: Wen, ZhaoHai, Xie, XinRan, Aleem, Muhammad Tahir, Aimulajiang, Kalibixiati, Chen, Cheng, Liang, Meng, Song, XiaoKai, Xu, LiXin, Li, XiangRui, Yan, RuoFeng
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 20.12.2021; BioMed Central; BMC
• Subjects: Animals; Bacteria; Biosynthesis; Cell culture; Cell Proliferation; Cloning, Molecular; Cytokines; Cytokines - genetics; Cytokines - metabolism; Developmental stages; Disease control; Drug dosages; Eggs; Enzymes; Female; Gene Expression Regulation - immunology; Gene Expression Regulation, Enzymologic; Genetic aspects; Goats; Gonads; Growth factors; Haemonchus - classification; Haemonchus - enzymology; Haemonchus - genetics; Haemonchus contortus; Immunofluorescence; Immunomodulation; Interleukin 10; Interleukin 17; Interleukin 4; Interleukins; Larvae; Leukocytes (mononuclear); Leukocytes, Mononuclear - physiology; Male; Microbial enzymes; Nematodes; Nitric oxide; Nucleotide sequence; Parasites; Parasitism; Pathogens; PBMCs; PCR; Peripheral blood mononuclear cells; Phosphatase; Phosphatases; Phosphates; Phosphoric Monoester Hydrolases - genetics; Phosphoric Monoester Hydrolases - metabolism; Phylogenetics; Phylogeny; Physiological aspects; Proliferation; Protein Conformation; Proteins; Rats; Recombinants; Reproducibility of Results; Secretion; Stat3 protein; Transcription; Transducers; Transforming growth factor-b; Trehalose; Trehalose-6-phosphate; Trehalose-6-phosphate phosphatase; Vaccines; China; Australia; United States > US; Haemonchus contortus; Trehalose-6-phosphate phosphatase; Immunomodulation; PBMCs
• ISSN: 1756-3305; 1756-3305
• DOI: 10.1186/s13071-021-05115-4

Trehalose-6-phosphate phosphatase (TPP6) is a key enzyme in the trehalose biosynthesis pathway. The accumulation of TPP6 inside the body is harmful to the pathogen, but almost nothing is currently known about the function of TPP6 from Haemonchus contortus (CRE-GOB-1). The H. contortus CRE-GOB-1 (HcGOB) gene was cloned and recombinant protein of GOB (rHcGOB) was expressed; transcription of the HcGOB gene at different developmental stages of H. contortus was then studied. The spatial expression pattern of the HcGOB gene in adult female and male worms was determined by both quantitative real-time PCR (qPCR) and immunofluorescence. The binding of the rHcGOB protein to goat PBMCs was assessed by immunofluorescence assay. The immunomodulatory impacts of rHcGOB on cell proliferation, nitric oxide generation and cytokine secretion were assessed by co-culture of rHcGOB protein with goat PBMCs. The HcGOB protein was transcribed in eggs, infective third-stage larvae (iL3s) and adults of H. contortus, with the highest transcript levels found in the egg stage. The transcript levels were significantly elevated in iL3s after manual desheathing. HcGOB was widely distributed in adult worms where it was mainly localized in the gut and gonads. rHcGOB was observed to bind to PBMCs and also to be recognized by sera collected from a goat infected with H. contortus. rHcGOB significantly activated the interleukin-10/transforming growth factor β/signal transducer and activator of transcription 3 (IL-10/TGF-β/STAT3) pathway in PBMCs while suppressing the transcription and expression of IL-4 and IL-17. These results suggest that the HcGOB gene plays an important role in the development, parasitism and reproduction of H. contortus. The rHcGOB protein affected the immunomodulatory function of PBMCs in the in vitro study, suggesting that this protein would be a promising vaccine target.