rBanLec, inside a positive dose-dependent manner, promotes the proliferation of TGMs from both BALB/c and C57BL/6 mice, while its mitogenic influence on RMs is significantly lower (BALB/c mice) or not detectable (C57BL/6 mice). Echinacoside of rBanLec binding to proteins in TGMs lysate. Whole cell lysate was prepared after collecting TGMs from peritoneum, resolved on 9% polyacrylamide gel by non-reducing SDS-polyacrylamide gel electrophoresis and transferred onto PVDF membrane. Binding of biotin-labeled rBanlec to the proteins from TGMs lysate was visualized with extrAvidine-alkaline phosphatase / 5-Bromo-4-chloro-3-indolyl phosphate/NBT system.(TIF) pone.0172469.s003.tif (365K) GUID:?D2EB01D1-AE74-4FEF-803C-AC11979A9CE0 S4 Fig: Western blot detection of TLR2 (A), TLR4 (B) and CD14 (C) in cell lysate prepared from peritoneal TGMs. Whole cell lysate was prepared after collecting ZCYTOR7 TGMs from peritoneum, resolved on 9% polyacrylamide gel by non-reducing SDS-polyacrylamide gel electrophoresis and transferred onto PVDF membrane. TLR2, TLR4 and CD14 were recognized using the specific biotin-labeled monoclonal antibodies. extrAvidine-alkaline phosphatase /5-Bromo-4-chloro-3-indolyl phosphate/nitro-blue tetrazolium chloride system was utilized for visualization.(TIF) pone.0172469.s004.tif (387K) GUID:?2A19BC6B-F632-4F2E-952B-0D6E7920E24B Data Availability StatementAll relevant data are within the paper and its Supporting Information documents. Abstract We shown that a recombinant banana lectin (rBanLec), which structural characteristics and physiological effects highly resemble Echinacoside those reported for its natural counterparts, Echinacoside binds murine peritoneal macrophages and specifically modulates their practical characteristics. By using rBanLec in concentrations ranging from 1 g to 10 g to activate resident (RMs) and thioglycollate-elicited (TGMs) peritoneal macrophages from BALB/c and C57BL/6 mice, we have shown that effects of rBanLec activation depend on its concentration but also within the practical status of macrophages and their genetic background. rBanLec, inside a positive dose-dependent manner, promotes the proliferation of TGMs from both BALB/c and C57BL/6 mice, while its mitogenic influence on RMs is definitely significantly lower (BALB/c mice) or not detectable (C57BL/6 mice). In all peritoneal macrophages, irrespective of their type and genetic background, rBanLec, inside a positive dose dependent manner, enhances the secretion of IL-10. rBanLec activation of RMs from both BALB/c and C57BL/6 resulted in a positive dose-dependent promotion of proinflammatory phenotype (enhancement of NO production and IL-12 and TNF secretion, reduction of arginase activity). Positive dose-dependent skewing toward proinflammatory phenotype was also observed in TGMs from C57BL/6 mice. However, the enhancement of rBanLec activation promotes skewing of TGMs from BALB/c mice towards anti-inflammatory profile (reduction of NO production and IL-12 secretion, enhancement of arginase activity and TGF and IL-4 secretion). Moreover, we founded that rBanLec binds oligosaccharide constructions of TLR2 and CD14 and that obstructing of signaling via these receptors significantly impairs the production of TNF and NO in BALB/c macrophages. Since the end result of rBanLec activation depends on rBanLec concentration as well as within the practical characteristics of its Echinacoside target cells and their genetic background, further studies are needed to investigate its effects under physiological and specific pathological conditions. Intro Macrophages represent large, morphologically and Echinacoside functionally heterogeneous, immune cell populace that is often regarded as a bridge between innate and adaptive immunity [1, 2]. As part of innate immunity, they provide a first line of defense against invading pathogens because of the inherent capabilities to phagocyte pathogens and mount various anti-microbial mechanisms. Instead, by acting as antigen-presenting cells, they exert an important part in initiation and shaping of the adaptive immune response [3]. The mode of macrophages activation, their earlier immunological encounter and surrounding milieu highly influence their practical characteristics [4C8]. Moreover, genetic background-dependent qualitative and quantitative variations in the macrophage reactions to external stimuli are reported [9]. Depending primarily on their morphological characteristics, peritoneal macrophages, most often used like a model system in macrophages-related practical studies, are divided into two organizations assigned as large peritoneal macrophages (LPMs) and small peritoneal macrophages (SPMs). Beside different morphology, a difference in phenotype and practical characteristics of LPMs and SPMs was designated as well [8, 10]. Even though SPMs are generally considered as more responsive to proinflammatory activation, it is hard to associate a specific type.