Recombinant protein of Haemonchus contortus 14-3-3 isoform 2 (rHcftt-2) decreased the production of IL-4 and suppressed the proliferation of goat PBMCs in vitro

Gadahi, Javaid Ali; Ehsan, Muhammad; Wang, Shuai; Zhang, Zhenchao; Wang, Yujian; Ren, Yan; Song, Xiaoling; Xu, Lixin; Li, Xiangrui

doi:10.1016/j.exppara.2016.10.014

Cited by 20 publications

(34 citation statements)

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“…Th17 effectors cells, capable of producing IL-17 cytokine, are associated with inflammatory responses and pathogenesis of various parasites [42, 43]. In accordance with our previous studies of HcESPs, rHcFTT-2 and rHcES-24 on cytokines production [41, 44, 45], it was suggested that, rHc-AK protein also participated in negative development of Th2 responses on IL-10 production (Fig. 5) and also involved in inflammatory reactions, favorable for parasite pathogenesis and survival by producing IL-17 in goat PBMCs.…”

Section: Discussionsupporting

confidence: 84%

“…Loke et al [53] reported that immune responses in helminths infection could be altered by regulating the proliferation of immune cells. In our previous research, we found a suppressive regulation of HcESPs, rHcFTT-2 and rHcES-24 on the cell proliferation in vitro [41, 44, 45]. Similarly, in the current study, rHc-AK could significantly inhibit the cell proliferation in goat PBMCs (Fig.…”

Section: Discussionsupporting

confidence: 79%

“…Coomes et al reported that IFN-γ could inhibit the development of Th2 immune responses [48]. In our previous study, the rHcFTT-2 protein increased the IFN-γ production by goat PBMCs in vitro [45]. In current research IFN-γ production was increased in goat PBMCs incubated with different rHc-AK concentrations (Fig.…”

Section: Discussionsupporting

confidence: 51%

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Arginine kinase from Haemonchus contortus decreased the proliferation and increased the apoptosis of goat PBMCs in vitro

et al. 2017

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BackgroundArginine kinase (AK), an important member of phosphagen kinase family has been extensively studied in various vertebrates and invertebrates. Immunologically, AKs are important constituents of different body parts, involved in various biological and cellular functions, and considered as immune-modulator and effector for pro-inflammatory cytokines. However, immunoregulatory changes of host cells triggered by AK protein of Haemonchus contortus, a parasitic nematode of ruminants, are still unknown. The current study was focused on cloning and characterisation of Hc-AK, and its regulatory effects on cytokines level, cell migration, cell proliferation, nitric oxide production and apoptosis of goat peripheral blood mononuclear cells (PBMCs) were observed.MethodsThe full-length sequence of the Hc-AK gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and sub-cloned into the prokaryotic expression vector pET-32a. The biochemical characteristics of recombinant protein Hc-AK, which was purified by affinity chromatography, were performed based on the enzymatic assay. Binding of rHc-AK with PBMCs was confirmed by immunofluorescence assay (IFA). Immunohistochemical analysis was used to detect localisation of Hc-AK within adult worms sections. The immunoregulatory effects of rHc-AK on cytokine secretions, cell proliferation, cell migration, nitric oxide production and apoptosis were determined by co-incubation of rHc-AK with goat PBMCs.ResultsThe full-length ORF (1080 bp) of the Hc-AK gene was successfully cloned, and His-tagged AK protein was expressed in the Escherichia coli strain BL21. The recombinant protein of Hc-AK (rHc-AK) was about 58.5 kDa together with the fused vector protein of 18 kDa. The biochemical assay showed that the protein encoded by the Hc-ak exhibited enzymatic activity. Western blot analysis confirmed that the rHc-AK was recognised by the sera from rat (rat-antiHc-AK). The IFA results showed that rHc-AK could bind on the surface of goat PBMCs. Immunohistochemically, Hc-AK was localised at the inner and outer membrane as well as in the gut region of adult worms. The binding of rHc-AK to host cells increased the levels of IL-4, IL-10, IL-17, IFN-γ, nitric oxide (NO) production and cell apoptosis of goat PBMCs, whereas, TGF-β1 levels, cell proliferation and PBMCs migration were significantly decreased in a dose dependent manner.ConclusionsOur findings suggested that rHc-AK is an important excretory and secretory (ES) protein involved in host immune responses and exhibit distinct immunomodulatory properties during interaction with goat PBMCs.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-017-2244-z) contains supplementary material, which is available to authorized users.

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Section: Discussionsupporting

confidence: 84%

Section: Discussionsupporting

confidence: 79%

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Arginine kinase from Haemonchus contortus decreased the proliferation and increased the apoptosis of goat PBMCs in vitro

et al. 2017

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“…The total RNA was extracted as previously described 25 . Briefly, about 100 adult worms were ground in pre‐chilled pestle and mortar with 1‐mL TRIzolfor 30 minutes.…”

Section: Methodsmentioning

confidence: 99%

“…24 In recent studies, various binding proteins of H contortus were evaluated for their immune-modulatory properties on goat PBMCs in vitro. [25][26][27][28][29] However, the immune functional roles of H contortus EF-1α in interaction with host immune cells are still unknown.…”

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confidence: 99%

Recombinant elongation factor 1 alpha of Haemonchus contortus affects the functions of goat PBMCs

Ehsan

Gadahi

et al. 2020

Parasite Immunology

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Excretory/secretory proteins of Haemonchus contortus (HcESPs) intermingle comprehensively with host immune cells and modulate host immune responses. In this study, H contortus ES antigen named as elongation factor 1 alpha (HcEF‐1α) was cloned and expressed. The influences of recombinant HcEF‐1α on multiple functions of goat peripheral blood mononuclear cells (PBMCs) were observed in vitro. Immunoblot analysis revealed that rHcEF‐1α was recognized by the serum of goat infected with H contortus. Immunofluorescence analysis indicated that rHcEF‐1α was bound on surface of PBMCs. Moreover, the productions of IL‐4, TGF‐β1, IFN‐γ and IL‐17 of cells were significantly modulated by the incubation with rHcEF‐1α. The production of interleukin IL‐10 was decreased. Cell migration, cell proliferation and cell apoptosis were significantly increased; however, nitric oxide production (NO) was significantly decreased. The MHC II molecule expression of cells incubated with rHcEF‐1α was increased significantly, whereas MHC‐I was not changed as compared to the control groups (PBS control and pET32a). These findings indicated that rHcEF‐1α protein might play essential roles in functional regulations of HcESPs on goat PBMC and mediate the immune responses of the host during host‐parasite relationship.

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Proteomic analysis of the excretory-secretory products from Strongyloides venezuelensis infective larvae: new insights for the immunodiagnosis of human strongyloidiasis

et al. 2022

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Recombinant protein of Haemonchus contortus 14-3-3 isoform 2 (rHcftt-2) decreased the production of IL-4 and suppressed the proliferation of goat PBMCs in vitro

Cited by 20 publications

References 61 publications

Arginine kinase from Haemonchus contortus decreased the proliferation and increased the apoptosis of goat PBMCs in vitro

Arginine kinase from Haemonchus contortus decreased the proliferation and increased the apoptosis of goat PBMCs in vitro

Recombinant elongation factor 1 alpha of Haemonchus contortus affects the functions of goat PBMCs

Proteomic analysis of the excretory-secretory products from Strongyloides venezuelensis infective larvae: new insights for the immunodiagnosis of human strongyloidiasis

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