title: 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.
abstract: 14-3-3 proteins have been found to be an excreted/secreted antigen and assumed to be released into the host-parasite interface and described in several unicellular and multicellular parasites. However, little is known about the immunomodulatory effects of H. controtus 14-3-3 protein on host cell. In present study, 14-3-3 isoform 2 gene, designated as Hcftt-2, was amplified by reverse transcription-polymerase chain reaction (RT-PCR) from the adult H. contortus cDNA and cloned into expression plasmid pET32a (+) and expression of the recombinant protein (rHcftt-2) was induced by IPTG. Binding activity of rHcftt-2 to goat peripheral blood mononuclear cells (PBMCs) was confirmed by immunofluorescence assay (IFA) and modulatory effects on cytokine production, cell proliferation, cell migration and nitric oxide (NO) production were observed by co-incubation of rHcftt-2 with goat PBMCs. Sequence analysis showed that it had significant homology with the known 14-3-3 protein isoform 2. Results of IFA revealed that, the rHcftt-2 was bound to the cell surface. We found that, the productions of IL10, IL-17, IFN-γ and cell migration of PBMCs were increased after the cells were incubated with rHCftt-2. However, the productions of IL-4, NO and cell proliferation of the PBMCs were significantly decreased in dose depended manner. Our results showed that the Hcftt-2 played important suppressive regulatory effects on the goat PBMCs.
authors.affiliation: College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, PR China. Electronic address: drgadahi@yahoo.com.
authors.forename: Javaid Ali
authors.initials: JA
authors.lastname: Gadahi
authors.affiliation: College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, PR China. Electronic address: mehsan124@gmail.com.
authors.forename: Muhammad
authors.initials: M
authors.lastname: Ehsan
authors.affiliation: College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, PR China. Electronic address: tongbaiws1003@163.com.
authors.forename: Shuai
authors.initials: S
authors.lastname: Wang
authors.affiliation: College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, PR China. Electronic address: zhangzhenchao525@163.com.
authors.forename: ZhenChao
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, PR China. Electronic address: 453425665@qq.com.
authors.forename: Yujian
authors.initials: Y
authors.lastname: Wang
authors.affiliation: College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, PR China. Electronic address: yanruofeng@njau.edu.cn.
authors.forename: RuoFeng
authors.initials: R
authors.lastname: Yan
authors.affiliation: College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, PR China. Electronic address: songxiaokai@njau.edu.cn.
authors.forename: XiaoKai
authors.initials: X
authors.lastname: Song
authors.affiliation: College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, PR China. Electronic address: xulixin@njau.edu.cn.
authors.forename: LiXin
authors.initials: L
authors.lastname: Xu
authors.affiliation: College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, PR China. Electronic address: lixiangrui@njau.edu.cn.
authors.forename: XiangRui
authors.initials: X
authors.lastname: Li
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000909:Antibodies, Helminth / Q000096:biosynthesis; D049109:Cell Proliferation / Q000187:drug effects; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D018509:DNA, Helminth / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005260:Female; D006041:Goats; D006189:Haemonchus / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D015801:Helminth Proteins / Q000737:chemistry / Q000276:immunology* / Q000494:pharmacology; D015847:Interleukin-4 / Q000276:immunology* / Q000378:metabolism; D009504:Neutrophils / Q000276:immunology* / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D020033:Protein Isoforms / Q000276:immunology / Q000494:pharmacology; D018510:RNA, Helminth / Q000235:genetics / Q000302:isolation & purification; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000276:immunology* / Q000494:pharmacology; D016415:Sequence Alignment
chemical_list: D000909:Antibodies, Helminth; D018076:DNA, Complementary; D018509:DNA, Helminth; D015801:Helminth Proteins; D020033:Protein Isoforms; D018510:RNA, Helminth; D011994:Recombinant Proteins; D015847:Interleukin-4; D009569:Nitric Oxide
keywords: 14-3-3 protein; Cytokine; Goat; Haemonchus contortus; PBMCs