Two basic proteins were isolated to homogeneity from malting barley (Hordeum vulgare L.) grain. Proteins were identified as members of a Thaumatin-Like Protein (TLP) family, by Western blot. Isoforms, assigned as TLP-S and TLP-R, have slightly different mobility at about 22 and 27 kDa in nonreducing and reducing conditions, and pI values of 9.5 and 9.4, respectively. The antifungal potency of malting barley grain TLPs isoforms was examined on Micrococcus lysodeikticus, Saccharomyces cerevisiae, Candida albicans and plant pathogen Fusarium sporotrichioides growth in vitro. It was found that that IC 50 value for TLP-S was two fold higher. Antibacterial and antifungal activities of both isoforms were completely abolished by divalent (Ca 2+ , Mn 2+ , Mg 2+ ) and monovalent (K + ) cations, at concentrations approximating physiological ionic strength and higher. Glucanase activity was not observed; neither TLP-S nor TLP-R digested glucan. On the basis of these results, the importance of TLP for barley grain protection against fungal diseases has been discussed together with the mechanism of antimicrobial action.
We demonstrated that a recombinant banana lectin (rBanLec), which structural characteristics and physiological impacts highly resemble those reported for its natural counterparts, binds murine peritoneal macrophages and specifically modulates their functional characteristics. By using rBanLec in concentrations ranging from 1 μg to 10 μg to stimulate resident (RMs) and thioglycollate-elicited (TGMs) peritoneal macrophages from BALB/c and C57BL/6 mice, we have shown that effects of rBanLec stimulation depend on its concentration but also on the functional status of macrophages and their genetic background. rBanLec, in 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). In all peritoneal macrophages, irrespective of their type and genetic background, rBanLec, in a positive dose dependent manner, enhances the secretion of IL-10. rBanLec stimulation 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 TNFα 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 stimulation 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 TGFβ and IL-4 secretion). Moreover, we established that rBanLec binds oligosaccharide structures of TLR2 and CD14 and that blocking of signaling via these receptors significantly impairs the production of TNFα and NO in BALB/c macrophages. Since the outcome of rBanLec stimulation depends on rBanLec concentration as well as on the functional characteristics of its target cells and their genetic background, further studies are needed to investigate its effects under physiological and specific pathological conditions.
Selective targeting of drugs to the proposed site of action provides therapeutic advantages such as reduced toxicity and smaller dose levels. Despite a huge progress made in drug design and delivery systems, many challenges still have to be solved. Small therapeutic drugs always have the potential to pass into the kidneys and be excreted from the body. The use of macromolecular constructs (carriers) that allow longer circulation times, contribute to improved chemical functionality and more precise drug delivery is an attractive alternative option. Bioadhesive systems which will utilize intense contact to increase the drug concentration gradient could be an attractive approach. Because of their specific carbohydrate-binding, lectins can interact with glycoconjugates present on the epithelial cells that line all of the organs exposed to the external environment. The unique carbohydrate specificities of plant lectins can facilitate mucoadhesion and cytoadhesion of drugs. As immunostimulatory molecules with an adjuvant effect plant lectins can also be employed in vaccine development
Art v 1 is the major allergen of mugwort (Artemisia vulgaris) pollen, a significant cause of hay fever all over Europe. Specific immunotherapy is the only treatment modality for allergic disease. Application of modified allergens makes the treatment safer and more efficient. In this work, two out of three (citraconic anhydride, cis-aconitic anhydride, 2,3-dimethylmaleic anhydride) tested anhydrides were proven to be suitable for chemical modifications of allergens. Art v 1 was modified by cis-aconitylation and citraconylation in order to obtain derivatives of Art v 1 that may be suitable for further immunological testing. Acylation of Art v 1 gave derivatives (caaArt v 1 and citArt v 1) with about 80 % modified amino groups. The derivatives were in the monomeric form and had dramatically reduced pI values. Both derivatives were relatively stable at neutral pH values, while the acyl groups undergo hydrolysis under acidic conditions. Modification of allergens by cis-aconitylation and citraconylation could be a new tool for obtaining allergoids.
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