[The human lectin supersystems possessing probiotic and protective actions.]

The potential of useful for human immunobiological supersystems of lectins (SSL) recognizing carbohydrates and glycoconjugates of molecular or supramolecular protein/(oligo)peptide-containing constituents of biotopes is described. SSL recognize, reversibly bind, delivery to biotopes, orient natural or synthetic polymeric, polyvalent glycoconjugates (imitators of natural glycopolymers) at the cell surface. The key features of SSL are indicated and described. The possibilities of application and prospects of SL of probiotics, complement С4 system and protein hormones (on example of erythropoietins) in prognostics and diagnostics of pathologies, prophylaxis and therapy of diseases and medical biotechnology are evaluated.

[Lectin populations of NK cells against tumors coupled to viral infections (review of literature).]

Analysis of the human NK (natural killers) cells and their functionally different populations in connection to tumor processes accompanied with viral infections is presented. Receptor lectins (non-immunoglobulin proteins and their complexes recognizing polysaccharides, glycoconjugates and glycopattern-containing molecules) play important role in regulation of innate immunity. Communicative diversity of NK-cell populations (in which lectins cofunction to other receptors) is directed against tumors and viruses. Effectiveness and selectivity of action of NK cell populations can be increased in cooperation together with adaptive immunity. Evaluations of occurrence, redistribution (also under influence of cytokines) and contribution of NK-populations (depending on lectin receptors recognition coupled to multifunctions of receptors) in respect of increasing antitumor and antiviral immune responces are given. The data indicate extended prospects of lectin receptors (coupled to other type receptors) containing NK populations of the network compartment of innate immunity upon realization of different variants of organism protection in cooperation with cellular and humoral immunity. Such NK populations are the basis for further intercellular interactions. Innate immunity Cross-Talk, involving the leader NK cell populations acting according to humoral immunity mechanisms, acts on duty regime (importance for therapy of chronic pathology) that results in providing optimal combined antitumor and antiviral cytokine and cytotoxic responses according to the principle of action as «network-in-network¼. The influence network of lectin, Ig-like, cytotoxic, other regulator NK populations (also throuph redistribution of production of cytokines by immunocompetent cells) is perspective for forming early prolongated antitumor and antiviral processes of different types in organism. It is of importance to consider CD diversity of receptor repertuar of lectin, Ig-like and other NK populations revealing different ontogenesis as well as to seach patient key NK-populations to select and construct personally (or for contingents in cases of epidemiological significance) optimal therapeutic/prophylactic NK populations (like variants of CAR-T). Aforementioned data indicate perspectiveness of NK cell populations in development of new antitumor/antiviral effective and selective vaccine strategies, preparations and regimes of their applications. Probiotic lectins reveal features of perspective ligands cofunctioning to network of NK cell populations.

[Lectins and glycoconjugates in presentation of antigens and protection against patogens.]

The current data on using lectin receptors and glycoconjugates (GC) in study of pathways of intercellular communications within innate immunity network coupled to adaptive immunity wеre overviewed and systematized. A superfamily of GC-specific C-type lectin receptors (CLR), as a constituent of pattern recognizing receptors (PRR), involves in immunity to produce redistribution of groups of cytokines, chemokines and specific antibodies. CLR on surface of multiple subpopulations of antigen-presenting cells (APC) and other cells of immunity are under consideration (monocyte derived macrophages, macrophages, immature and mature dendritic cells, other myeloid cells, neutrophils, basophils, lymphocytes, etc.). CLR cofunction both each other and/or together with other PRR and immune receptors. CLR are favourable for quicker and GC-specific innate type presentation of important danger antigenic patterns (PAMP, MAMP, DAMP, SAMP, TAMP) using their specific targeting through currently/ adaptively optimal pathways within intercellular branched network of activated/ polarized subpopulations of cells mentioned above. CLR agonist GC (recombinant, synthetic, conjugates to carriers (adaptive proteins, etc.), assembled into nanoparticles and liposomes) can serve successful and specific adjuvants in vaccine strategies. CLR and GC pairs participate in on duty biocontrol of immunobiological supervising in organism, modulate/ switch recognition and protection cell pathways of immune responses (presentation, reciprocally organized protective attacks, rates and amplitudes of activation and inhibition of cell communicative chains). Lectins characterized both cell and cell communicative chain current functional biomarkering that is of importance for clinical diagnostics. The knowledge on CLR-GC pattern relationships is perspective for study and influence fine mechanisms of cell GC-targeting and immunity, improvement of traditional vaccines, development of GC-specific and highly effective pattern vaccines against different infections or tumors. New specific vaccine strategies are possible towards different biotopes in organism (also for mucosal opened cavities).

[The role of lectins from probiotic microorganisms in sustaining the macroorganism].

The notion of lectins has been expanded with special reference to lectins of probiotic microorganisms (LPM). New data on LPM and their properties are considered in the context of a new concept of systemic organization of LPM being developed. Conditions for complete separation of poly (N-acetylgalactosamine-?)-binding lectins and polyMan-?/oligomannoside/mannan-binding lectins are developed on the assumption that they contain a single carbohydrate-binding site in their structure. It is shown that strain typing of probiotic microorganisms is possible based not only on protein but also on LPM composition. The observed peculiarities of LPM functioning substantially extend the concept of informational superiority of integral interatomic networks over their isolated components. This approach makes it possible to set functioning of LPM systems against that of lectin systems of other components of human microbiota as prevailing in the support of the normal status of the macrorganism. The results demonsatrate the unique character of LPM systems and suggest good prospects for their application in medicine, biotechnology and nanotechnology.

[Classification of lectins as universal regulator molecules of biological systems].

Modern concepts of lectin classification reflecting tendencies in the development of functional classification of lectins (Ln) and Ln-like proteins are presented. Classification of bacterial Ln is proposed. Classification of Ln and Ln-like proteins of plants, animals, and bacteria is based on 8 major elements of protein secondary structure that allows for the evaluation of topographic diversity of their surfaces, potential compatibility of bacterial, vegetable, and mammalian Ln molecules and possible variants of their co-functioning.

[Nanotechnologies and perspectives of their application in medicine and biotechnology].

The results of nanotechnologies application in biological and medical fields are summarized. The examples of nanotechnologies use in reactions of cell adhesion, cell recognizing, intracellular sorting, organ and tissue construction, nanobiosensing, drug delivering and cancer therapy are presented. The prospects of nanotechnologies use in biomedicine, biothechnology and cell biology are shown.

[General properties and functioning principles of lectins in biological systems].

The authors give the literature review with analysis of main conceptions of lectins - their structure and functions according to state-of-the-art science and production requirements. Up-to-date definition of lectins based on last achievements of science and practice represented in the article.

[Strategic aspects of constructing probiotics of the future].

The publication systematizes modern achievements in understanding the role of microbiocenoses in the existence of macroorganism, and possible ways of constructing of probiotics of the future.

[Deficiencies in C4A and C4B isotypes of human complement revealed by isoelectrofocusing and chemical reactivity of activated forms].

An approach is proposed to detect deficiencies in isotypes A and B of the C4 component of human complement, based on the calculation of the ratio of their IEA activities and the ratio of their quantities determined by isoelectrofocusing of their desialated forms with chemiluminescent detection in an immunoblot. The ratios of the quantities and activities of C4A/C4B practically coincided when determined in blood serum of 20 patients, many of which had inherited deficiencies in the C4 component isotypes.

[New generation pharmaceuticals from milk of transgenic animals. The problem of isolation of biologically active proteins].

The authors of the review characterize the protein components of milk and adduce data on the expression of transgenic proteins in animal milk. Technological principles of purification of recombinant medicinal proteins from milk of transgenic animals are covered.

[Biological characterization and prospects of application of a new bifidobacterial strain as an ingradient of probiotics].

The study was dedicated to investigation of a bifidobacterial strain, isolated from a healthy chick, its biological properties and lectins produced by the strain; a restrictive analysis of the chromosomal DNA was conducted as well. Presumably B. gallinarum was identified. The strain displayed adhesive, lectinic and antagonistic activities; it was resistant to the leading chemotherapeutic pharmaceuticals and antibiotics, and, when given to chicks, it had probiotic effect. The study identified a set of lectins binding phosphorylated mannan, with isoelectric points in the pH interval of 4 to 8; the set differed from the sets of lectins of other bifidobacteria.

[Lectins, adhesins, and lectin-like substances of lactobacilli and bifidobacteria].

Cell-surface adhesion factors of lactobacilli and bifidobacteria, such as lectin/adhesin proteins of S-layers, secreted lectin-like bacteriocins, and lectin-like complexes, are considered and classified in the article. Certain general and specific properties of these factors are noted, such as in vitro and in vivo adhesion, cell co(aggregation), participation in the forming of microbial biofilms and colonization of mammalian alimentary tract, as well as complexation with biopolymers and bioeffectors, specificity to glycanes and natural glycoconjugates, domain and spatial organization of adhesion factors, co-functioning with other cytokines (pro- and anti-inflammatory ones), regulation of target cell properties, and other biological and physiological activities. The authors also note possibilities of application of lectins and lectin-like proteins of probiotic strains of lactobacilli and bifidobacteria in medicine and biotechnology.

[Formation of probioticotherapy in Russia].

Probioticotherapy and probioticoprophylaxis have been applied widely in Russia and the world of late. The article presents the main medical, strain-related, and biotechnological stages of their development in this country, in comparison with the results of the world practice. The authors substantiate appropriateness of creation of probiotics and which would contain either only growth medium components, able to activate normoflora (to increase colonization ability and stimulate production of biologically active substances) upon entering the organism, or only probiotic strain components and/or biologically active substances produced by biologically active strains in mono- or associative cultivation, or combination of the listed components.

[The effects of the microbial components of the probiotic Acilact on the cell-mediated immunity factors under experimental conditions].

The authors analyzed the effects of the microbial components of the probiotic Acilact on the cell-mediated factors of the immune homeostasis. According to the results, L. acidophilis stimulates the functional activity of immunocompetent cells, especially T-cell mediated immunity with strain- and dose-depending differences in intensity. The study demonstrated the ability of L. acidophilis to activate expression of the genes of pro-inflammatory cytokines (IFNgamma, IL-6, 12, and TNFalpha), to induce production of IFNalpha and IFNgamma, and increase interferon production, with additional stimulation of lymphoid organ cells with specific inductors in vitro. The study established that L. acidophilis stimulates production of macrophage migration inhibition factor by immunocompetent cells of Peyer's plaques and the splen, and induces its quantitative increase in the serum of CBA mice. The study detected immunocorrecting effect of L. acidophilis by the example of experimental shigellous infection, demonstrated intensification of the prolipherative activity of immunocompetent T-cells, induction of the expression of the genes of pro-inflammatory cytokines (IFNgamma, IL-1beta, 6, 12, and TNFalpha), but not IL4 and 10, increase of synthesis of IFNalpha and IFNgamma by the cells of Peyer's plaques, the splen and thymus. The obtained results prove the immunological effect of the probiotic preparation Acilact, based on the strains L. acidophilis NK1 and L. acidophilis K3(111)24.

[Isolation and some properties of a lectin from the venom of the Vietnamese scorpion Buthus occitanus sp]. / Vydelenie i nekotorye svoistva lektina iz iada V'etnamskogo skorpiona Buthus occitanus sp.

A lectin was isolated from the venom of scorpion Buthus occitanus sp. by means of Sephadex G-50 gel filtration and CM-cellulose ion exchange chromatography. The homogeneous lectin preparation consisted of homodimeric molecules with a subunit Mr of 9.3 kDa. Glycine, alanine, and serine dominated in the lectin amino acid composition. The lectin was a glycoprotein containing 20% carbohydrates (predominantly mannose and glucose). Trypsin-treated murine erythrocytes agglutinated at the lectin concentration of 32 micrograms/ml. Hemagglutination was inhibited by carbohydrates (L-fucose > D-glucose > L-rhamnose > D-xylose). The lectin revealed no phospholipase or hyaluronidase, nor toxic activity.

[Inborn deficiency of complement C4A and C4B isotypes in persons infected with Chlamydia]. / Vrozhdennye defitsity izotipov C4A i C4B komponenta C4 komplementa u lits, inifitsirovannykh khlamidiiami.

The difference in the functional activity of the isotypes A and B of component C4 of human complement was used to determine their ratio to detect the inherited deficiency of the isotypes. The frequency of deficiency in healthy persons blood donors was equal for C4A and C4B (0.14 for each isotype), i.e. 14% of total number (22) donors, or 28% totally. These results agree with the literary data, on which the frequency of deficiency of C4A is 0.14, and of C4B is 0.11-0.16. The inherent deficiencies of C4A and C4B for persons infected by Chlamydia were studied. For this purpose the patients (35 persons) with in blood antibodies (IgG or IgM) to Chlamydia (C. trachomatis, C. psittaci and C. pneumoniae) were investigated. The frequencies of deficiency of C4A and C4B were 0.29 and 0.46 respectively. Thus, the number of the undeficiency patients was only 25%, while among healthy persons 70-75% of individuals not having deficiencies of isotypes C4 were observed. The deficiencies of isotypes of C4 at this pathology is detected for the first time. The obtained data suggest the existence of the predisposition to the development of diseases stipulated by Chlamydia in persons with inherent deficiency of C4 component of complement.

[Isotyping of human C4 complement using differences in the functional activity of C4A and C4B isotypes]. / Izotipirovanie komponenta C4 komplementa cheloveka s ispol'zovaniiem razlichii v funktsional'noi aktivnosti isotipov C4A i C4B.

The difference in the functional activity of the isotypes A and B of component C4 of human complement was used to determine their ratio and to detect the inherited deficiency of the isotypes. ELISA methods were developed for the quantitative assay of component C4 (conventional sandwich method) and its functional activity. When determining the functional activity, the classic pathway of the complement and therefore of component C4 was activated by activators sorbed on ELISA microplates (immunoglobulin IgG3 or liposaccharide of the Shigella sonnei cell walls, which activates the complement by binding component C1). The nascent fragment C4b is covalently bound to the target activator; C4Ab binds better to the target protein (immunoglobulin), and C4Bb to the target carbohydrate (liposaccharide). Therefore, when immunoglobulin is a target activator, isotype C4A is bound and determined; and when the complement is activated by liposaccharide, isotype C4B is determined. The ratio of the activities determined by the two methods indicates a deficiency in the individual isotypes of component C4 or its absence. The rabbit polyclonal monospecific antibodies against the human component C4 and the conjugates of these antibodies with horseradish peroxidase were used in the methods described.

[Inhibition of binding of activated compliment component C4b with its target]. / Ingibirovanie sviazyvaniia aktivirovannogo C4b-komponenta komplementa s mishen'iu.

The inhibition of covalent binding of the nascent C4b fragment of the human complement component to its natural target, immunoglobulin G, was studied. To this end, an immunoenzyme system was developed. In this ELISA method, the complement was activated on the sorbed IgG molecules and the resulting nascent C4b fragment acylated IgG or interacted with a competitive inhibitor added to the system. The inhibition constants for binding of the nascent C4b to its target were determined for immunoglobulins G1, G2, G3, G4, M, and A1, as well as for ferritin, yeast mannan, capsid polysaccharides of the Neisseria meningitidis A, B, and C serotypes, diphtheria anatoxin, epinephrine, and salicylic acid. On the basis of the experimental data, the immunoglobulin role at the activation stage of the complement regulation cascade, the relationship between the antigen immunogenicity and its ability to interact with C4b, and the direct effect of a number of therapeutic agents on the complement system were discussed. Lectins of various specificities were shown to inhibit the enzymic activation of C4 by the first complement component and the subsequent C4b sorption to its target, which allowed us to suggest that some oligosaccharide fragments of the C1s and C4 molecules are spatially close to the C1s active site and to the thioester bond of C4.

[Isolation and characteristics of a lectin-like protein from bee venom]. / Vydelenie i kharakteristika lektinopodobnogo belka iz pchelinogo iada.

Lectin-like properties of bee venom proteins were studied. Phospholipase A2 was demonstrated to harbor lectin activity. Specific hemagglutinating activity of the isolated phospholipase exceeded 320 times the activity of the initial bee venom.

[An affinity chromatography sorbent containing concanavalin A groups immobilized by complex formation with cobalt]. / Affinnyi khromatograficheskii sorbent, soderzhashchii gruppirovki konkanavalina A, immobilizovannye kompleksoobrazovaniem s kobal'tom.

A Spheron-base affinity sorbent containing Concanavalin A (Con A) groupings has been synthesized. Con A was immobilized via the formation of a ternary complex with a stationary ligand (tetrafunctional) triethylene tetramine) and cobalt(III) ions. The inertness of cobalt(III) provided strong binding of Con A to the matrix. The sorbent was tested for the ability to absorb proteins (albumin, lysozyme, heparin, and ribonuclease) and used the isolation of glucosaminidase from the proteolytic complex of the fungus Geotrichum candidum. The purified enzyme was characterized biochemically.