Purification and Characterization of a Mannose Recognition Lectin from Oreochromis niloticus (Tilapia Fish): Cytokine Production in Mice Splenocytes

Abstract

The aim of this work was to purify and partially characterize a mannose recognition lectin from Nile tilapia (Oreochromis niloticus) serum, named OniL. OniL was isolated through precipitation with ammonium sulfate and affinity chromatography (Concanavalin A-Sepharose 4B). In addition, we evaluated carbohydrate specificity, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) profiles, and in vitro immunomodulatory activity on mice splenocyte experimental cultures through cytotoxic assays and cytokine production. The ammonium sulfate fraction F2 showed the highest specific hemagglutinating activity (331) and was applied to affinity matrix. Adsorbed proteins (OniL) were eluted with methyl-α-D: -mannopyranoside. OniL, a 17-kDa protein by SDS-PAGE constituted by subunits of 11 and 6.6 kDa, showed highest affinity for methyl-α-D: -mannopyranoside and D: -mannose. Immunological assays, in vitro, showed that OniL did not show cytotoxicity against splenocytes, induced higher IFN-γ production and lower IL-10 as well as nitrite release. In conclusion, OniL lectin was successfully purified and showed a preferential Th1 response in mice splenocytes.

Anti-biofilm properties and immunological response of an immune molecule lectin isolated from shrimp Metapenaeus monoceros

“…The analysis of purified MmLec was multivalent showing higher affinity to arabinose, dextrose, rhamnose, N-acetyl glucosamine, fucose and mannose, a lower affinity to N-acetyl galactosamine, and no activity towards galactose. This agrees with previous results by Silva et al [63] and Mitra and Das [64] who found that a single lectin protein can exhibit affinity towards various sugar molecules.…”

The Deposition of a Lectin from <em>Oreochromis niloticus</em> on the Surface of Titanium Dioxide Nanotubes Improve the Cell Adhesion, Proliferation, and Osteogenic Activity of Osteoblast-like Cells

“…In this work, we used the lectin OniL to functionalize the surface of TiO2NTs. This lectin is a 17 kDa protein consisting of two subunits of 11 and 6.6 kDa and presents a high affinity for methyl-α-Dmannopyranoside and D-mannose [14]. In mammals, mannose-binding lectins (MBLs) constitute a fundamental link between the innate immune system and other functions, such as coagulation, homeostasis after injury and defence against microorganisms [35].…”

“…Lectins, a non-immune proteins, which bind reversibly and in a highly specific manner to simple or complex carbohydrates, have been widely investigated due to its prominent role in several physiological and pathological processes, including immune response, inflammation, cell-cell communication, recognition, and differentiation [8]. It has been demonstrated that lectins from plants and animals have mitogenic [9,10], antibacterial [11,12], immunomodulatory [13,14] antithrombotic [15] and healing [16] activities. Some of these activities are remarkably interesting from the point of view of osseointegration and tissue repair.…”

Lectins, Interconnecting Proteins with Biotechnological/Pharmacological and Therapeutic Applications

“…A mannose-specific lectin from serum of Oreochromis niloticus (Nile tilapia fish) denominated OniL demonstrated in vitro Th1 response induction on mice splenocytes. OniL also stimulated high levels of IFN γ production and low levels of IL-10 and nitrite, without cytotoxic effect [65]. …”