Abstract

Haemonchus contortus has developed complexed and multifaceted mechanisms of immune evasion to enable the survival in the host. Generating excretion and secretion products (ESPs) to subvert or suppress the functions of host cytokines is a newly immune regulatory pattern found during recent years. Transforming growth factor-β (TGF-β) has critical immune regulatory functions in nematode infections. In this study, co-immunoprecipitation (co-IP) assay was used to identify the goat TGF-β1 binding proteins from HcESPs. The interaction between TGF-β1 and nematode fatty acid retinoid binding domain containing protein of H. contortus (HcFAR) was analyzed by glutathione S-transferase (GST)-pull down assay. The suppressive effect of rHcFAR on TGF-β1-induced immunoglobulin A (IgA) secretion was observed by co-incubation of rHcFAR and TGF-β1 with goat peripheral blood mononuclear cells (PBMCs). The IgA concentrations were determined using enzyme linked immunosorbent assay (ELISA) kit. Meanwhile, the suppressive effect of rHcFAR on TGF-β1-induced T helper (Th) 9 differentiation was investigated by co-incubation of rHcFAR, TGF-β1 and interleukin (IL)-4 with goats PBMCs. In parallel, IL-4 was replaced by IL-6 to determine the effects on the Th17 differentiation. The transcriptions of IL-9 and IL-17 in PBMCs were then evaluated by real-time PCR. Finally, we found that HcFAR from HcESPs could bind to goat TGF-β1 in vitro. The ELISA results of IgA showed that 40 μg/mL rHcFAR could suppress the IgA secretion of PBMCs induced by TGF-β1. Additionally, rHcFAR (at 10 μg/mL and 20 μg/mL) could inhibit the mRNA transcription of IL-9 induced by TGF-β1 and IL-4. Meanwhile, rHcFAR could also downregulate the transcription of IL-17 induced by TGF-β1 and IL-6 in a dose-dependent manner. These results indicated that HcFAR was a functional inhibitor of goat TGF-β1 and this information may help contribute to understanding of the relationship between the ESPs and host cytokines.

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