Signaling events during macrophage activation with Betula pendula roth pectic polysaccharides.

We studied the effect of two pectic polysaccharides PS-B1-AG and PS-B2-RG that were contained in total polysaccharides extracted from Betula pendula leaves on NO production by mouse macrophages and the contribution of signaling molecules to macrophage activation by the test substances. Unlike the total sample, pectins produced a NO-stimulating effect on macrophages. The effect of PS-B2-RG (10 µg/ml) did not differ from the effect of LPS, while PS-B1-AG produced this effect only in a concentration of 20 µg/ml, which was probably due to differences in the chemical structure of the test substances. The studied pectin polysaccharides activated transcription factor NF-κB, kinases p38 and PI3, and cAMP as a negative regulator. These results indicate that Betula pendula polysaccharides are promising substances for creation of immunomodulating drugs.

Effect of IKK-2 inhibitor on local Th1 and Th2 type inflammation.

We studied the capacity of IKK-2 inhibitor ([5-(p-fluorophenyl)-2-ureido] thiophene-3-carboxamide) and non-steroidal anti-inflammatory drug voltaren to reduce the severity of local inflammation in Th1- and Th2-type immune response. The test substances (inhibitor in concentrations from 2×10(-13)to 2×10(-16)M and 9.8×10(-4)M voltaren) were shown to reduce edema caused by sheep erythrocytes (Th1-type immune response) and local immediate-type hypersensitivity response (Th2-type immune response). The anti-inflammatory effects of IKK-2 inhibitor in Th1-type immune response were similar to those of voltaren.

Effect of NF-κB inhibitor aurothiomalate on local inflammation in experimental Th1- and Th2-type immune response.

We compared the effect of NF-κB inhibitor aurothiomalate and voltaren on local inflammation in different types of immune response. Both substances reduced edema caused by sheep erythrocytes (Th1-type immune response) and local immediate-type hypersensitivity response induced with ovalbumin (Th2-dependent response). The anti-inflammatory effects of aurothiomalate were similar to those of voltaren during Th1-type immune response.

In vitro effect of combined hybrid molecules from vitamin E analogues and betulinic acid on macrophage activity.

Macrophage activity was studied after treatment with hybrid molecules obtained by condensation of terpenic acid residues (betulinic and betulonic acids) and α-tocopherol analogues (α-tocopherol hemisuccinate and Trolox acid). As distinct from betulinic acid and α-tocopherol hemisuccinate, hybrid molecules did not exhibit cytotoxicity in relation to mouse peritoneal macrophages in the MTT test. Test substances inhibited the production of NO by mouse peritoneal macrophages. However, hybrid molecules had no effect on activity of macrophage arginase. Our results indicate that new molecules have anti-inflammatory activity. It can be hypothesized that these substances have immunomodulatory properties.

Study of anti-inflammatory action of aurothiomalate, an inhibitor of NF-κB.

We compared the effects of NF-κB inhibitor aurothiomalate and voltaren on NO production by mouse macrophages in vitro, their ability to cause local edema at the site of injection, and their effect on carrageenan-induced inflammation. High concentrations of aurothiomalate reduced NO production, while in low concentrations both aurothiomalate and voltaren stimulated this process. When injected into mouse footpad, aurothiomalate in a dose >1 mM and voltaren in a dose >1.6 µM induce paw edema. Both compounds suppressed carrageenan-induced inflammation, but the efficacy of aurothiomalate 2-fold exceeded that of voltaren.

Water-soluble polysaccharide obtained from Acorus calamus L. classically activates macrophages and stimulates Th1 response.

A pectic polysaccharide was isolated from the rhizomes of Acorus calamus L. The main component of its carbohydrate chain was represented by residues of D-galacturonic acid (>85%). In addition, this polysaccharide contained residues of galactose, arabinose, xylose and rhamnose (<10%). Structural analysis of pectic polysaccharide from A.calamus L. with NMR spectroscopy indicated that it contains the regions of a linear 1,4-alpha-D-galactopyranosyluronan, which represents a major component of the macromolecule. A considerable amount of galacturonic acid residues was not methoxylated. We demonstrate here that the pectic polysaccharide from A. calamus L. in low concentrations was able to stimulate in vitro IL-12 and nitric oxide production by murine macrophages. It also induced TNF-alpha secretion by human peripheral blood mononuclear cells, reduced arginase activity but did not affect IL-10 secretion by murine macrophages or human peripheral blood mononuclear cells. The IL-12 and NO-stimulating effects on murine macrophages were similar to that of LPS. In addition, the polysaccharide promoted in vivo Th1 immune response in mice which were immunized with sheep red blood cells (DTH and quantity of plaque-forming cells) and down regulated serum level of IgG1 and IgE during Th2-depend immune response induced by ovalbumin. The PS increased Th1-induced edema and suppressed Th2-induced paw swelling in adoptive systems. Our results suggest that the pectic polysaccharide from A. calamus L. represents a promising immunomodulating agent that stimulates M1-polarized macrophages and promotes Th1-oriented adaptive immune response. We propose that this polysaccharide could be potentially applied for treatment of infectious, oncological diseases or for immunoglobulin-E-mediated disorders.

Effects of plant water-soluble polysaccharides on the production of immunoglobulins E and G1 by lymphocytes of mice sensitized with ovalbumin.

Were studied the effects of water-soluble plant polysaccharides isolated from pharmacopoeic raw material on anaphylactic shock and production of IgE and IgG1 by lymphocytes from mice immunized with ovalbumin. Course treatment with polysaccharides from coltsfoot, sweet flag, clover, Artemisia, marigold, and elecampane reduced animal mortality after induction of anaphylactic shock. In addition, injection of these substances reduced serum concentrations of IgE and IgG1. These substances can be regarded as promising agents for the treatment of IgE-dependent diseases (atopic dermatitis, asthma, atopic rhinitis, urticaria, food allergies, etc.).