The pervasive effects of recombinant Fasciola gigantica Ras-related protein Rab10 on the functions of goat peripheral blood mononuclear cells

Tian, Ai-Ling; Lu, MingMin; Zhang, Fu-Kai; Calderón-Mantilla, Guillermo; Petsalaki, Evangelia; Tian, XiaoWei; Wang, WenJuan; Huang, Si-Yang; Li, XiangRui; Elsheikha, Hany M.; Zhu, Xing-Quan

Abstract

Background: Fasciola gigantica-induced immunomodulation is a major hurdle faced by the host for controlling infection. Here, we elucidated the role of F. gigantica Ras-related protein Rab10 (FgRab10) in the modulation of key functions of peripheral blood mononuclear cells (PBMCs) of goats.
Methods: We cloned and expressed recombinant FgRab10 (rFgRab10) protein and examined its effects on several functions of goat PBMCs. Protein interactors of rFgRab10 were predicted in silico by querying the databases Intact, String, BioPlex and BioGrid. In addition, a total energy analysis of each of the identified interactions was also conducted. Gene Ontology (GO) enrichment analysis was carried out using FuncAssociate 3.0.
Results: The FgRab10 gene (618 bp), encodes 205-amino-acid residues with a molecular mass of ~23 kDa, had complete nucleotide sequence homology with F. hepatica Ras family protein gene (PIS87503.1). The rFgRab10 protein specifically cross-reacted with anti-Fasciola antibodies as shown by Western blot and immunofluorescence analysis. This protein exhibited multiple effects on goat PBMCs, including increased production of cytokines [interleukin-2 (IL-2), IL-4, IL-10, transforming growth factor beta (TGF-β) and interferon gamma (IFN-γ)] and total nitric oxide (NO), enhancing apoptosis and migration of PBMCs, and promoting the phagocytic ability of monocytes. However, it significantly inhibited cell proliferation. Homology modelling revealed 63% identity between rFgRab10 and human Rab10 protein (Uniprot ID: P61026). Protein interaction network analysis revealed more stabilizing interactions between Rab proteins geranylgeranyltransferase component A 1 (CHM) and Rab proteins geranylgeranyltransferase component A 2 (CHML) and rFgRab10 protein. Gene Ontology analysis identified RabGTPase mediated signaling as the most represented pathway.
Conclusions: rFgRab10 protein exerts profound influences on various functions of goat PBMCs. This finding may help explain why F. gigantica is capable of provoking recognition by host immune cells, less capable of destroying this successful parasite.

Citation

Tian, A.-L., Lu, M., Zhang, F.-K., Calderón-Mantilla, G., Petsalaki, E., Tian, X., …Zhu, X.-Q. (2018). The pervasive effects of recombinant Fasciola gigantica Ras-related protein Rab10 on the functions of goat peripheral blood mononuclear cells. Parasites and Vectors, 11, Article 579. https://doi.org/10.1186/s13071-018-3148-2

• Journal Article Type: Article
• Acceptance Date: Oct 14, 2018
• Online Publication Date: Nov 6, 2018
• Publication Date: Nov 6, 2018
• Deposit Date: Nov 7, 2018
• Publicly Available Date: Nov 7, 2018
• Journal: Parasites & Vectors
• Print ISSN: 1756-3305
• Publisher: BioMed Central
• Peer Reviewed: Peer Reviewed
• Volume: 11
• Article Number: 579
• DOI: https://doi.org/10.1186/s13071-018-3148-2
• Keywords: Parasitology; Infectious Diseases
• Public URL: https://nottingham-repository.worktribe.com/output/1233210
• Contract Date: Nov 7, 2018