[Elaboration of immune stimulating lipid-saponin subunit antigen carrier based on glycolipid monogalactosyldiacylglycerol from sea macrophytes and triterpene glycosides from Cucumaria japonica].

The ability of some triterpene glycosides of holothurians: holotoxin A1 from Apostichopus japonicus and a mixture of monosulphated triterpene glycosides from Cucumaria japonica called cucumarioside (CD) to form supramolecular complexes with cholesterol (Chol) and monogalactosyldiacylglycerol (MGDG) or phosphatidylcholine (PC) was studied. A transmission electron microscopy method was used to observe supramolecular lipid-saponin complexes formed by holotoxin A1 and CD with cholesterol in the presence of membrane lipids. The observed supramolecular complexes are tubular nanoparticles with a length of 100-300 nm, an external diameter of 10-16 nm and an internal diameter of 2-6 nm. The formation of tubular nanoparticles was more effective in the presence of MGDG than with PC. Nanoparticles forming in the presence of MGDG are shaped as a tubule, have a constant diameter and a strongly pronounced internal channel. In contrast, PC has no such properties; this lipid is unable to fully integrate in tubular nanoparticles. Based on electron-microscopy data the range of weight ratio of MGDG-Chol-CD was determined as a 1-10:2:3 that provided most effective formation of tubular nanoparticles. Different methods of incorporation of model antigens in complex MGDG-Chol-CD were studied. Influenza haemagglutinin and neuraminidase from commercial vaccine "Influvac" and pore forming protein YompF from Yersinia pseudotuberculosis were used as model antigens. From 54 to 72% of protein of "Influvac" vaccine and 88-92% of YompF were incorporated in supramolecular complexes depending on the method of incorporation. The loss of functional activity of haemagglutinin of vaccine "Influvac" was the result of applying ultrasonic disintegration for incorporation of this protein in complex MGDG-Chol-CD. YompF incorporation in MGDG-Chol-CD complex led to the increased diameter of tubular particles, in the same time incorporation of vaccine "Influvac" antigens produced the "cap" formation at the end of tubules. The possibility of a described supramolecular complex MGDG-Chol-CD to be a carrier for subunit bacterial and viral antigens is shown.

[Enhancing the immunogenic activity of influvac vaccine in the use of adjuvant TI complexes modified by echinochrome A].

The self-assembly of marine macrophyte glycolipids, holothurian saponin, and cholesterol gave rise to nanoscale morphological structures called tubular immunostimulating (TI) complexes. Whether the latter could be used on the basis of vaccine preparations containing the influenza virus subunit antigens was studied. There was an obvious increase in the immunogenicity of influenza virus hemagglutinin when the experimental animals were immunized with this antigen as part of TI complexes. It was shown that the adjuvant activity of the TI complex to influenza virus hemagglutinin could be enhanced by adding the known antioxidant echinochrome A from a sand-dollar (Echinarachnius parma) to the matrix of the TI complex.

[The comparative study of the medical action of lyuteolin, rosmarinic acid and echinochrom A at experimental stress-induced cardiopathology].

The method of the physical load in condition of the coronary circulation of the blood disturbance, caused mesaton injection, induced development rat cardiopathology, bring about of the heart function decompensation and 40% death of experimenthal animals. Under electronic-microscopic study of rat cardiomyocytes are discovered signs to disorganizations of mitochondrial apparatus of these cells. Administration to therapeutic mode of luteolin and echinochrome A preparations has provided to 100% animal probability of survival. At the same time, mitochondrial apparatus of cardiomyocytes was characterized by the normal parameter i.e. given preparations have provided of defensive adative effect at cardiomyocytes level. Similar activities for rosmarinic acid have not shown. Study some metabolic parameter and endocrine status animal has also allowed revealing of therapeutic effect of luteolin and echinochrome A. Findings be evidence of that echinochrome A and luteolin capable to play the important positive role in metabolism of cardiomyocytes by stimulating of mitochondrial biogenesis and by changing of adaptative mechanisms of the organism cardiovascular system protection.

[Immunogenic and protective properties of nanosized constructs based on tubular immunostimulating complexes and pore forming protein of Yersinia pseudotuberculosis].

AIM: Evaluation of immunogenic and protective properties of constructs based on subunit porin antigen from Yersinia pseudotuberculosis, immunostimulating complexes (ISCOM) and tubular immunostimulating (TI) complexes. MATERIALS AND METHODS: Porin antibodies and blood serum cytokines were determined by using EIA. Porin-specific cell immunity was evaluated by DTH reaction inflammation index. Protective activity of porin formulations was determined by measuring specific gravity of animals surviving Yersinia pseudotuberculosis lethal challenge. RESULTS: Porin in TI complexes develops higher immunogenicity when compared with individual protein or protein with complete Freunds adjuvant. Porin in TI complexes develops higher protective activity, inhibits interferon synthesis in mice. Incorporation of porin into TI complexes results in neutralization of porin suppressive activity against DTH mechanisms and interferon system. CONCLUSION: TI complexes may be used as perspective carriers for bacterial antigens. TI complexes have adjuvant properties and can provide protective properties to porin vaccine constructs.

[Complexation between triterpene glycosides of holothurians and cholesterol is the basis of lipid-saponin carriers of subunit protein antigens].

The ability of some triterpene glycosides of holothurians: cucumarioside A2-2 from Cucumaria japonica, cucumarioside G1 from Cucumaria fraudatrix, frondoside A from Cucumaria frondosa, and holotoxin A1 from A postichopus japonicus to form lipid-saponin supramolecular complexes was studied. The formation of supramolecular cholesterol-glycosides complexes between cholesterol and these glycosides in water medium was observed by transmission electron microscopy. These complexes were considered as nanoparticles with different structure. Complexes formed by cholesterol with cucumarioside A2-2, holotoxin A1, and frondoside A are tubular nanoparticles. In contrast, complexes between cholesterol and cucumarioside G1 have different structured. The structure of nanoparticles formed in the presence of cucumarioside A2-2, holotoxin A1, and cucumarioside G1 was dependent on the ratio of cholesterol in the lipid-saponin system. On the other hand, frondoside A did not shown this tendency. In lipid-saponin systems with a similar molar ratio cholesterol-glycoside, the ordering of the supramolecular structure decreases in the following order: cucumarioside A2-2, holotoxin A1, frondoside A. A comparative analysis of the morphology of the supramolecular complexes and the peculiarities of the molecular structure of triterpene glycosides studied, demonstrated that the structure of supramolecular complexes formed depends on the branching and length of the glycoside carbohydrate chain. On the other hand, the formation of monomeric cholesterol-glycosides complexes depends on the peculiarities of the structure of aglycone. Thus, the possibility of the formation of a new type of antigen carries on the basis of marine triterpene glycosides was proved.

[Membranotropic effect of some triterpene glycosides possessing immunostimulating properties].

The peculiarities of the interaction between cell membrane lipids and triterpene glycosides from holothurians Apostichopus japonicus S. and Cucumaria japonica (holotoxin A1 and cucumarioside A2-2, respectively) were studied in comparison with plant saponins from Quillaja saponaria, known as hemolytic, adjuvant, and structure-forming components of immunostimulating complexes. Similar to Quillaja saponins, the sea glycosides, holotoxin A1 and cucumarioside A2-2 were shown to possess a high hemolytic activity (2.6 and 3 microg/ml, respectively) and sterol-depending membranotropic effect mediated by the formation of nonbilayer sterol-lipid-glycoside complexes. At the same time, cucumarioside A2-2 bound exogenic cholesterol only in the presence of membrane lipids, such as phosphatidylcholine or monogalactosyldiacylglycerol, in contrast to Quillaja saponins and holotoxin A1, which bound cholesterol in the molar ratios 1:2 and 1:8, respectively. Moreover, in all cases, tree-component complexes containing cholesterol, lipid, and glycoside exhibited a lower hemolytic activity compared with two-component sterol-glycoside complexes. It was concluded that the hydrophobic medium of cell membranes performs a potentiative role in the effective interaction between triterpene glycosides and "sterol receptors". A method for decreasing the toxicity of membranotropic holothurian glycosides possessing the immunomodulating properties was suggested.