Abstract
Galectins are glycan-binding proteins that are widely expressed and distributed in mammalian tissues as well as cells of innate and adaptive immune responses. CD4+ T-helper cells differentiate into effector subsets in response to cytokines. T helper 9 cells are one of the recently described subsets of effector T cells that are relatively new and less studied. In this study, galectin domain containing protein from Haemonchus contortus (Hc-GDC) was cloned, expressed in pET32a, and immunoblotting was performed. Localization of recombinant (r)Hc-GDC on outer and inner surface of H. contortus worm and binding with goat Peripheral Blood Mononuclear cells (PBMCs) were performed using immunofluorescence assay. Moreover, effects of rHc-GDC on proliferation, apoptosis, cell migration, and the nitric oxide production in goat PBMCs were evaluated. Furthermore, modulatory effects of rHc-GDC on production of Th1, Th2, and Th9 cells were evaluated by flowcytometry and on interferon gamma, interleukin (IL)-4 and IL-9 were evaluated by quantitative real-time polymerase chain reaction. The results demonstrated that rHc-GDC was successfully cloned, expressed in expression vector as well as in the gut surface of adult H. contortus worm and successful binding with PBMCs surface were observed. Immunoblotting results revealed that rHc-GDC is an important active protein of H. contortus excretory and secretory products. Moreover, the interaction of rHc-GDC with host cells increased the production of Th2, Th9 cells, IL4, IL-9, PBMC proliferation, nitric oxide, and cell migration. No effects of rHc-GDC were observed on PMBC apoptosis, production of Th1 cells, and secretions of IFN-γ and IL-10 cytokines. These findings indicate that recombinant GDC protein from H. contortus modulates the immune functions of goat PBMCs and has the potential to enhance protective immunity by inducing T helper-9-derived IL-9 in vitro.
Highlights
Galectins, animal lectins, are known to be widely distributed from lower invertebrates, such as nematodes and sponges, to higher vertebrates [1]
Recombinant Galectin domain containing protein (GDC) protein from H. contortus was characterized and localization of this protein by immunofluorescence assay confirmed the binding with host immune cells, in surface ligand complex shape, which is a characteristic feature of Excretory and Secretory Products (ESPs) to modulate the immune functions of host Peripheral Blood Mononuclear cells (PBMCs) [47]
This study revealed that rHc-GDC is an important active protein of H. contortus ESPs that played crucial roles in the immune regulations
Summary
Animal lectins, are known to be widely distributed from lower invertebrates, such as nematodes and sponges, to higher vertebrates [1]. It has been demonstrated that the interaction of galectin with its receptor on the target cell governs the function of the galectin [2]. Galectins are widely involved in mammalian innate and adaptive immune responses [3,4,5,6,7]. All galectins contain conserved carbohydrate-recognition domains (CRDs) of about 130 amino acids that are responsible for carbohydrate binding and may influence cellular functions through protein to protein interactions with other nuclear and cytoplasmic proteins. Galectins play important roles in diverse pathological and physiological processes, including immune and inflammatory responses, by maintaining T cell homeostasis [8]. The major function of the immune system is to detect and abolish pathogenic parasites, microorganisms, and cancer cells [9]. Galectin-like proteins have been identified in Onchocerca volvulus [11], Teladorsagia circumcincta [12], and Haemonchus contortus [13]
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