Promoting agent of proliferation of regulatory t-lymphocyte, and their promotion method
SUBSTANCE: invention proposed N-end fragment of soluble suppressor of immune response with the length of 21 amino-acids, which has sequence of amino-acids as per Seq ID NO: 1, allowing to promote formation of regulatory T-lymphocytes, as well as a promotion method of formation of regulatory T-lymphocytes with N-end fragment of soluble suppressor of immune response with Seq ID NO: 1, at its introduction with the concentration of 0.1-50 microgram/ml.
EFFECT: invention can be used for propagation of regulatory T-lymphocytes obtained from a patient suffering autoimmune disease, for the purpose of further injection of the obtained T-lymphocytes into the body of the patient.
3 cl, 2 tbl, 3 ex
The invention relates to the field of biotechnology, biologics, medical devices, designed to impact on the immune system of the body, namely the activity of T-lymphocytes, namely a stimulator of proliferation of regulatory T-lymphocytes.
Minor subpopulation of T-lymphocytes called regulatory T lymphocytes (Treg), capable of using multiple mechanisms to inhibit the proliferation of normal T-lymphocytes, having an important role in suppressing the activity of T-cells directed against antigens of the organism. Phenotypic regulatory T-lymphocytes are distinguished by the presence of markers of CD4+CD25+FoxP3+ [Aune T.M. et al. // J. Immunol. - 1983. - v. 131. - p.2848-2852; Webb D.R. et al. // International Immunology. - 1990. -v.2. - p.765-774].
The high content of regulatory T-lymphocytes was observed in the lymphocytes of tumor and draining lymph nodes of tumor that is probably one of the reasons for the suppression of T-cell immune response of the body to the tumor antigens. In this regard, the increased activity and/or maintenance of Treg in the body may become the treatment of several autoimmune diseases, complications associated with transplantation of organs and tissues, including bone marrow transplantation, such as disease "graft versus host". In particular, in the present method of treatment of autoimmune and alloimmune disease is the second by a fence of a patient, in need of this treatment, leukocyte mass, the separation from her cell with the phenotype CD4+CD25+, further cultivation of these cells in vitro for up to 4 weeks with the purpose of reproduction and reverse the introduction of the received cells to the patient (WO 2005086781, 2004).
As agents stimulating the proliferation of regulatory T cells is suggested to use azacytidine, tumor necrosis factor-beta, retinoic acid, trichostatin a, anti-CD3, anti-CD28, oxidized ATP, interleukin-2 (WO 2009126877, 2008; WO 2009114097, 2008).
The disadvantage of these stimulants is that they are not strictly selective stimulators of proliferation of regulatory T-lymphocytes, causing the need to find a more specific activator of proliferation of such cells.
Closest to the claimed invention is the use as a stimulator of cytokines, particularly of interleukin-2, which is injected into the culture medium in a dose of 100 IU/ml (WO 2005086781, 2004). The disadvantage of this stimulant is the lack of specificity.
The problem faced by the authors, was expanding Arsenal of stimulators of proliferation of regulatory T cells on the proliferation of regulatory T-lymphocytes.
The technical result was achieved in the synthesis of peptide representing the N-terminal fragment of soluble su the spring immune response (SIRS 1-21), the amino acid sequence of which is presented in Annex 1, Seq ID NO: 1, which was having voting stimulating effect on the proliferation of regulatory T cells (Treg) in populations of lymphocytes in the thymus and spleen.
Protein SIRS (soluble immune response suppressor), the complete amino acid sequence of which is still unknown, is a polypeptide with a molecular weight of about 10-14 kDa, which is produced by CD8+ cells, activated mitogen, antigen or interferon (US 4771125, 1984) and has the ability to inhibit the production of antibodies and to inhibit the reaction of the delayed-type hypersensitivity in vivo [Aune T.M. et al. // J. Immunol. - 1983. - v. 131. - p.2848-2852; Webb D.R. et al. // International Immunology. - 1990. - v.2. - p.765-774]. The amino acid sequence of the N-terminal segment SIRS mouse, containing 21 amino acid residue (SIRS 1-21), set [Webb D.R. et al. // International Immunology. - 1990. - v.2. - R-774]. Information about the use of SIRS or its fragments as stimulators of proliferation of Treg cells in the reviewed literature was not found.
During the studies the authors found that the SIRS peptide 1-21 during culturing a population of cells containing T-lymphocytes that selectively stimulates the proliferation of regulatory T lymphocytes (Treg, CD4+CD25+FoxP3+ cells), practically no impact on the proliferation of total is opulatio cells, that may be associated with the inhibition of regulatory T-lymphocytes proliferation of normal lymphocytes.
Stimulation of the formation of regulatory T-lymphocytes is carried out by introducing the culture fluid stimulator SIRS 1-21 in a concentration of 0.1-10 µg/ml While the efficiency increases with the simultaneous introduction into the system of interleukin-1-beta at a dose of 100-300 PG/ml
The nature and advantages of the claimed group of inventions are illustrated by the following examples.
Example 1. Chemical synthesis of the N-terminal fragment of SIRS (SIRS1-21)
Synthesis of peptide SIRS1-21performed by a solid phase method on a synthesizer Vega Coupler 250 according to the method of in situ using Nα-Boc-protected derivatives of amino acids. In the work we used the following derivatives of amino acids Boc-Ser(Bzl)-OH, Boc-Thr(Bzl)-OH, Boc-Pro-OH, Boc-Ile-OH, Boc-Asn(Trt)-OH, Boc-Gln-OH, Boc-Glu(OBzl)-OH, Boc-Met-OH, Boc-Asp(OcHex)-OH, Boc-Ala-OH, Boc-Lys(ClZ)-OH. The source of the aminoacyl-polymer served as Boc-Ser(Bzl)-PAM with specific capacity of 0.64 mmol/g.
Release temporary tert-butyloxycarbonyl protection held 50% triperoxonane acid (TFA) in methylene chloride (DCM). The condensation reaction was carried out in dimethylformamide (DMF). Neutralization in the first condensation was carried out by adding a threefold excess diisopropylethylamine (DIPEA) directly into the reaction mixture at the stage of joining AMI is kislotno residue; re-condensation was carried out after an additional washing peptidyl-polymer 10% solution of DIPEA in DMF. The joining of amino acid residues was performed by the method of activated esters derived from the corresponding derivatives of amino acids using aminobutiramida-carbodiimide, using a 5-fold excess of reagent in DMF. The mixture of amino acid derivatives were prepared immediately before introduction into the reaction vessel. The completeness of the condensation reaction was performed using ninhydrin and bromophenol tests.
After completion of the extension of the peptide chain enters the polymer was treated with 50% TFA twice for 1 minute, washed with DCM and diethyl ether was removed from the reactor and dried to constant weight.
Cleavage of the peptide from the resin and removal of side protective groups was performed using liquid hydrogen fluoride on Snl mechanism in the presence of scavengers.
Selected coarse product was subjected to purification by the method of prepreparation reversed-phase HPLC on a column (Waters Prep Nova-Pak HR C-18, C, 60Å, 19×300 mm (detection at 220 nm).
The fraction corresponding to the peak of the main product, after freeze drying were subjected to mass spectral and HPLC-analysis.
According to MALDI-TOF spectrometry molecular weight of the peptide (M+N)+- 2281. theoretical molecular weight in the calculation of the free peptide - 2280,38
The content of the basic substance, the SIRS peptide1-21amounted to more than 95% according to HPLC.
Example 2. The influence of the SIRS1-21on the proliferation of splenocytes
The SIRS peptide1-21bred in the medium RPMI-1640 with 1% fetal calf serum at a concentration of 1 mg/ml, were prepared serial dilutions and added into the wells of 96-Loup night - tablet 50 ál per well. Later in the wells contributed suspension of splenocytes in 100 μl of RPMI-1640 medium containing 10% fetal calf serum (TES), 3×105cells per well. A production test was carried out not less than 4 Parallels for each experimental point.
The Board was placed in CO2incubator and incubated at 37°C in conditions of absolute humidity. After 48 h from the beginning of incubation in cell culture was made3H-thymidine (Isotope, St. Petersburg) at a final concentration of 5 mccu/ml Through day cells were transferred to glass fiber filters using a FilterMate harvester 96 (Perkin-Elmer) and determined the intensity of the incorporation of thymidine on a tablet β-counter MicroBeta TriLux 1450 (Perkin-Elmer).
The results, expressed in number of H3-thymidine incorporated into acid-insoluble residue (imp/min) are presented in table 1.
|In Janie SIRS peptide 1-21on the proliferation of mouse splenocytes in vitro|
|SIRS1-21µg/ml||The intensity of the proliferation of splenocytes (imp/min)|
Analysis of the results indicate a weak inhibition of the proliferation of splenocytes in the presence of 0.1 μg/ml of the investigated peptide. Lower and higher concentrations SIRS1-21during the analyzed period of time (1 day) effects on the proliferation of splenocytes did not.
Example 3. Assessing the impact of SIRS peptide1-21on the proliferation of regulatory T-cells in vitro.
During the experiment came from the fact that cytofluorimetric regulatory T cells can be identified so CD4+ CD25+ lymphocytes, also expressing the transcription factor FoxP3.
The thymocytes or splenocytes mouse were incubated with peptide SIRS1-21in the presence of 100-300 PG/ml recombinant IL-1β or without it for 3 days. To determine the percentage of regulatory T-cells in suspense and thymocytes and splenocytes cells were stained with monoclonal antibodies to surface markers CD4, CD8 and CD25, and intracellular marker FoxP3. Used antibodies to mouse eBioscience: anti-Rohr-FITC (clone FJK-16s), anti-CD4-PE (clone GK1.5), anti-CD25-PE-Cy5 (clone RS), anti-CD8a-PE-Cy5 (clone 53-6 .7). For staining with antibodies to FoxP3 used a set of Foxp3 / reduced Factor Staining Buffer Set (eBioscience) according to the manufacturer's instructions. Analysis of the expression of relevant markers was performed in a three-color mode on cytofluorimetry EPICS XL (Beckman-Coulter). The results obtained are presented in table 2.
|The influence of the SIRS1-25on the CD4+CD25+FoxP3+cells in cultured thymocytes and splenocytes mouse|
|SIRS1-21µg/ml||The CD4+CD25+FoxP3+ cells (%)|
|Without IL-1β||With IL-1β||3 PG/ml||Without IL-1β||With IL-1β||3 PG/ml|
|50||0,41||0,90||0,99||0,95||7,39||5,58||5,61||the ceiling of 5.60|
Table 2 shows that the best results are achieved by incubation of the cells in the presence of 16.5 ág/ml SIRS1-21. Thus, there is a 1.5 to 2-fold increase in the content of CD4+CD25+FoxP3+ in suspensions of thymocytes and 2-3-fold increase in the content of the cells of this phenotype in suspensions of splenocytes.
For thymocytes effect SIRS1-21on the CD4+CD25+FoxP3+cells is more pronounced on the background of IL-1β, which is itself on the content of the data cells in the population of thymocytes virtually no effect. Thus, SIRS1-21stimulates the proliferation of Treg in populations of thymocytes and splenocytes, including the presence in the environment of the cultivation of IL-1β.
From the above data it follows that the SIRS peptide1-21when culturing a population of cells containing T-lymphocytes that selectively stimulates the proliferation of regulatory T lymphocytes (Treg, CD4+CD25+FoxP3+cells), practically no impact on the proliferation of the total population of cells.
This property makes this peptide promising for breeding ex vivo regulatory T-lymphocytes obtained from a patient suffering from autoimmune disease associated with chronic inflammation, for example, a patient with rheumatoid arthritis, psoriasis and psoriaticescom arthritis, diabetes 1-g is of type autoimmune Hashimoto (Hashimoto's disease), ulcerative colitis, Crohn's disease, granulomatosis vasculitis, sarcoidosis, scleroderma, multiple sclerosis, systemic sclerosis, glomerulonephritis autoimmune nature and other similar ailments, graft rejection, diseases, graft versus host, with subsequent introduction of the obtained T-regulatory T-lymphocytes of this patient for treatment of the above diseases and pathological conditions.
1. The N-terminal fragment of soluble suppressor of the immune response length of 21 amino acid having the amino acid sequence according to Seq ID NO: 1, as a stimulant formation of regulatory T-lymphocytes.
2. Method of promoting the formation of regulatory T-lymphocytes by culturing cells containing T-lymphocytes, in the presence of the stimulator, wherein the stimulator into the culture medium injected N-terminal fragment of soluble suppressor of the immune response according to claim 1 in a concentration of 0.1-50 μg/ml
3. The method according to claim 2, characterized in that in the environment of the cultivation of additionally injected interleukin-1-beta at a dose of 100-300 PG/ml
SUBSTANCE: invention includes developed medium formulations for cultivation of human fibroblasts in presence of low (2%) concentration of serum of xenogenic, allogenic and autogenic (autologous) origin.
EFFECT: reduced variability of composition of substantial growth factors, risk of contamination with cell pathogens, reduced level of media cytotoxicity and their potential immunogenicity; creation of adequate low-serum media for maintennace and growth media for solutionof various tasks of diploid strains and tissue cultures cultivation; development of the possibility to use an autogenic serum for cultivation of cells and tissues of a donor with no risks of infectious contamination, cytotoxicity and immune conflict.
2 cl, 5 dwg, 4 ex
SUBSTANCE: method of reprogramming somatic mammalian cells is proposed, comprising bringing the mammalian somatic cell into contact with reprogramming factors selected from the group consisting of: (i) Oct4; (ii) Oct4 and Klf4; (iii) Oct4, Klf4 and Sox2, and with Wnt-conditioned medium or activator of the pathway Wnt.
EFFECT: invention can be used for obtaining induced pluripotent stem cells in order to treat or prevent a medical pathological state in an individual.
10 cl, 1 tbl, 5 dwg, 8 ex
SUBSTANCE: method enables assessing marrowy neutrophilopoietic cell reactivity on intercellular signals - cytokines generated by mature cells of the same type. In case of the suppression of differentiation and morphofunctional generation of neutrophilic precursors in the presence of a great number of neutrophilokines, the cells are stated to be healthy, otherwise - as pathological.
EFFECT: use of the declared method enables the relatively simple assessment of marrowy neutrophilopoietic cell activity.
SUBSTANCE: there are described methods for differentiation of pluripotent stem cells by culture of the pluripotent stem cells in a medium of certain chemical composition added with B27 and treated with activin A.
EFFECT: invention provides improved differentiation effectiveness in stem cells and pancreatic endocrine cells.
32 cl, 53 dwg, 7 tbl, 37 ex
SUBSTANCE: disclosed protein-free cell culture media contain mineral salts, amino acids, antioxidants, vitamins, nutrients, antibiotics, D-mannitol and methyl cellulose, having molecular weight of 14000 Da.
EFFECT: disclosed media can be used in assisted reproduction procedures, particularly extracorporeal fertilisation.
SUBSTANCE: described is a method of producing induced pluripotent stem cells (iPS-cells) from skin fibroblasts of a patient with Huntington's chorea, involving production and reproduction of fibroblasts, inoculating said fibroblats on pools with monolayer density 30% in a medium of DMEM+P/S+L-Glu+10% FBS+bFGF 2 ng/mcl, infection of cells when monolayer density reaches 60% with four viruses LeGO-hOct4-MOI 5, LeGO-hSox2-MOI 5, LeGO-hc-Myc-MOI 2.5, LeGO-hKlf4-MOI 5. After four days, the cells are re-inoculated on plastic 1 to 22.5 in a medium of DMEM+P/S+L-Glu+10% FBS; after one day, the medium is replaced with ESmed and cultured for 8-10 days; the medium is replaced once in two days; mechanical re-inoculation is carried out 1 to 1 on matrigel in a medium of mTeSRI; culturing is carried out until growth of columns of morphologically similar to embryonic stem cells; the clones are collected after a week. Disclosed is a cell line obtained using the described method, which is meant for treating Huntington's chorea, which has the following morpho-molecular characteristics: size of the order of 20 mcm, large nucleus, growth in compact colonies with close contact, doubling period as embryonic stem cells, considering error, normal kryotype of 46 chromosomes, expression of Oct4, SOX2, FOXD3, HESX1, Nanog, Sall4.
EFFECT: high treatment efficiency.
2 cl, 5 dwg, 2 tbl, 1 ex
SUBSTANCE: invention relates to animal husbandry. The medium for cryopreservation of goat sperm contains crystalline glucose, tris-(hydroxymethyl)-aminomethane, citric acid 1-hydrate, egg yolk, glycerin, distilled water, amber acid and sodium alginate, with the following ratio of components, wt %: crystalline glucose 0.6000-0.667; tris-(hydroxymethyl)-aminomethane 3.635-4.039; citric acid 1-hydrate 1.887-2.084; amber acid 0.197-0.232; sodium alginate 0.020-0.040; glycerin 5.000-5.500; egg yolk 2.500-3.000; distilled water - the rest.
EFFECT: invention improves stability of sperms to cryopreservation and stabilises their biological full-value in freezing-thawing processes.
7 tbl 16 ex
SUBSTANCE: nutrient medium contains KNO3, NH4NO3, CaCl26H2O, MgSO47H2O, KH2PO4, FeSO47H2O, Na2EDTA2H2O, H3BO3, MnSO44H2O, ZnSO47H2O, KI, Na2MoO42H2O, CuSO45H2O, CoCl26H2O, nicotinic acid, thiamine, pyridoxine, 2,4-D, benzylaminopurine, sucrose, agar-agar and water in the preset proportions.
EFFECT: invention allows culturing the SAUSSUREA ORGAADAYIV KUAN ET KRASNOB cell culture.
SUBSTANCE: group of inventions relates to medicine, namely to dentistry, and deals with medications for restoration of periodontal tissues and methods of their obtaining. Biotransplant for treatment of periodontal diseases represents suspension which contains cultures of autological or allogenic fibroblasts, or fibroblast-like cells in 0.9% solution of sodium chloride in concentration 2.0-10.0x106 in 1 ml of biotransplant, integrated on pharmaceutically acceptable biocompatible biodegradable carrier, selected from the group: acellular matrix, hyaluronic acid preparation, acellular matrix and hyaluronic acid preparation, with specified volume ratio of cell culture suspension ands carrier, which makes it possible to obtain consistence convenient for injecting and, together with expressed clinical effect ensures low traumaticity and simplicity of injecting. Preferable, as carrier, used is crushed to size 100-200 mcm acellular matrix "Saimetra". Also claimed is method of treating periodontal diseases, which lies in the following: on mucous membrane of gum in region of defect one week before biotransplant introduction preliminarily made are cuts provoking an acute inflammatory process resulting in increase of tissue volume, which makes it possible to carry out lossless introduction of large volume of biotransplant. After that, into defect area by means of injection two times with interval 1-4 weeks introduced is claimed biotransplant, using 3.6-60×106 cells per treatment course, which ensures optimal time interval for cell engraftment and restoration of periodontal tissues.
EFFECT: increase of treatment efficiency.
7 cl, 3 ex, 2 dwg
SUBSTANCE: cell material containing marrow stem cells in a liquid culture medium is placed over a prepared agar medium containing marrow cells producing homing factors. After incubation of the prepared two-layer culture, inadherent cells are transferred to a semi-viscous culture medium and incubated in a mode required for colony-formation. Then by recording the difference of stem cell count in the cell material in the reference, and after placing on the agar medium containing homing factors, stem cells migrated by stem cell homing factor are counted.
EFFECT: invention allows simplifying the method for determination of production of stem cell homing factors due to the use of the agar medium as a semipermeable membrane and the introduction of colony-forming ability change of the cell material as an evaluation criterion of production of stem cell homing factors.
1 tbl, 1 ex
SUBSTANCE: group of inventions refers to medicine and concerns using a polynucleotide which contains a sequence coding a C-terminal heavy subunit domain of tetanus toxin (HcTeTx), for treating amyotrophic lateral sclerosis (ALS), as well as using a polynucleotide which contains a C-terminal of a heavy subunit of tetanus toxin (HcTeTx), for treating the above disease.
EFFECT: group of inventions provides relieving the ALS symptoms ensured by introducing an isolated non-toxic fragment of tetanus toxin (HcTeTx or a fragment thereof) or the polynucleotide coding the above fragment.
17 cl, 4 ex, 11 dwg, 2 tbl
SUBSTANCE: group of inventions refers to methods of treating urinogenital neurological disorders with using drug preparations containing a modified clostridial toxin, and using modified clostridial toxins in preparing drug preparations for treating the urinogenital neurological disorders. The methods of treating the urinogenital neurological disorders involve the stage of administering a therapeutically effective amount of a composition containing the modified clostridial toxin into a mammal in need thereof.
EFFECT: group of inventions is characterised by the effective treatment of the urinogenital neurological disorders and minimal effects in the regions being other than the goals of the toxin therapy.
15 cl, 11 dwg, 2 tbl, 11 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: group of invention refers to medicine and concerns a pharmaceutical composition, and a method of stimulating an immune response to Mycobacterium avium, subspecies paratuberculosis (MAP) in a mammal. The composition contains a recombinant polypeptide containing from N-terminal to C-terminal: a C-terminal fragment of MAP_3527 protein, an amino acid sequence of MAP_1519 protein, and then an N-terminal protein of MAP 3527 protein. The method involves administering the pharmaceutical composition in animals in the amounts sufficient to stimulate the immune response on MAP.
EFFECT: group of inventions provides either eliminating, or delaying the MAP proliferation over a period of infection.
18 cl, 3 ex, 31 dwg, 2 tbl
SUBSTANCE: invention discloses a bispecific antibody or its functional fragment, specifically binding with IL-4 and IL-13, which contain variable domains of light and heavy chains with established amino acid sequence. The invention also includes use of antibodies or its functional fragments within a pharmaceutical composition for treatment of diseases or disturbances mediated by IL-4 and/or IL-13, including allergic diseases, asthma, cancer. The invention discloses a molecule of nucleic acid, which codes a bispecific antibody or its fragment, an expression vector and a master cell for production of a bispecific antibody and its functional fragment.
EFFECT: invention makes it possible to produce and use new inhibitors of cytokines, preserving stability in process of production and use in vivo.
17 cl, 2 dwg, 8 tbl, 8 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: inventions refers to biotechnology and concern hypoallergic fused proteins. A presented hypoallergic fused protein consists of one hypoallergic molecule originated from an allergen, fused with a second non-allergic protein originated from a pathogen, or a fragment thereof with the hypoallergic molecule originated from the allergen showing 50% decreased ability to bind IgE, 30% decreased T-cell responsiveness as compared with a wild-type allergen. A nucleic acid molecule coding the fused protein, an expression vector, a host cell expressing the same protein, and a vaccine composition containing the protein are also presented.
EFFECT: presented invention enables preparing therapeutic and prophylactic drugs for an allergy or diseases caused by pathogens with using no toxic adjuvants, showing no side effects.
14 cl, 23 dwg, 17 tbl, 27 ex
SUBSTANCE: group of inventions refers to medicine and aims at managing or preventing post-chemotherapeutic pain or pains induced by an anti-cancer agent. What is declared is the use of at least one type A botulinum neurotoxin for preparing a medicine for managing or preventing post-chemotherapeutic pain or pains. What is also declared is a method of managing or preventing post-chemotherapeutic pain or pains by local introduction of said medicine to achieve a generalised effect.
EFFECT: use of the declared group of inventions provides a high therapeutic effect.
2 ex, 3 dwg, 32 cl
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention discloses a number of polynucleotides and polypeptides β-hemolytic streptococci, in particular polypeptides and polynucleotides of Streptococcus pyogenes, and based on them immunogenic compositions, used for prevention or reduction of symptoms of colonisation or infection, caused by β-hemolytic streptococci. Immunogenic composition (versions) contains mixture of two or more polypeptides, encoded by sequence of nucleic acid (NA), which has, at least, 90% identity of sequence of NA, selected from group, consisting of peptidase C5a (SCP), open reading frame (OPC)554, OPC 1218, OPC 1358 and OPC 2459. One of versions of immunogenic composition contains polypeptide SCP, polypeptide peptidylpropylisomerase and, at least, one other polypeptide. Also disclosed are methods of protecting susceptible mammal against colonisation or infection, caused by β-hemolytic streptococcus, by immunisation of immunogenic composition by invention.
EFFECT: invention provides immunogenic compositions and methods for prevention or reduction of symptoms of infections, caused by β-hemolytic streptococci of group A, B, C and G, as well as ensures immunity to wide spectrum of bacteria BHS.
41 cl, 16 dwg, 2 tbl, 3 ex
SUBSTANCE: invention relates to biochemistry. Disclosed is a polypeptide essentially consisting of amino acid sequence SEQ ID NO: 23 for inducing immune response in a mammal against meningococcal bacteria. Provided is a nucleic acid which codes said polypeptide. The invention also provides a plasmid which contains a nucleotide sequence and a host cell which is transformed by said plasmid, which are meant for expressing said polypeptide. Provided is a membrane vesicle containing the polypeptide for use as a medicament for preventing meningococcosis in a mammal. Disclosed is an immunogenic composition which contains an effective amount of the polypeptide or vesicle.
EFFECT: invention enables to induce immune response in a mammal against meningococcal bacteria.
14 cl, 5 tbl
SUBSTANCE: invention relates to biotechnology, specifically obtaining immunogens from Neisseria meningitides and can be used in medicine. Disclosed is protein which is 80% or more identical to amino acid sequence SEQ ID NO:19, and an immunogenic composition with said protein.
EFFECT: invention enables to use said protein for effective prevention or treatment of bacterial meningitis.
6 cl, 5 dwg, 28 tbl
SUBSTANCE: invention discloses a purified and/recombinant antigen polypeptide possessing toxin activity, recovered from Clostridium perfringens with specified amino acid sequence. The invention discloses the recovered or recombinant polynucleotide coding such polypeptide, an expression vector and a host cell expressing the polypeptide. The invention discloses a method for preparing the polypeptide, an antibody specifically bound with the polypeptide, immunogenic compositions and vaccines containing the given polypeptide or a polynucleotide thereby providing a specifically immune response to the polypeptide. There are disclosed a method for inducing the immune response, a method of determining the fact whether an individual has been exposed to a pathogen (versions), a method of screening, an agonist or an antagonist modulating activity of the polypeptide, a method of animal vaccination, e.g. hens for inducing active immunity, as well as passive immunity in hen off-springs which becomes less sensitive to clostridial diseases. What is disclosed is a transgenic plant containing the exogenous polynucleotide coding the polypeptide under the invention, applicable for animal feeding.
EFFECT: polypeptide is used as an ingredient of a forage or a beverage for preventing a disease caused by bacteria expressing the polypeptide under the invention.
39 cl, 8 dwg, 6 tbl, 11 ex
SUBSTANCE: invention relates to medicine, namely to ophthalmology and can be applied for treatment of traumatic and dystrophic injuries of eye cornea. For this purpose peptide fragments of protein S100b - SP2 and/or SP3 in concentrations 10-6 M are introduced in conjunctive cavity or in endonasal way. Introduction is realised daily two times per day during 10 days.
EFFECT: method ensures fast and high-quality restoration of injured zone of cornea due to stimulation of regenerative-reparative processes in cornea.
3 ex, 2 cl