Cartridge genetic maker expressing three biologically active siphk, effectively attacking human immunodeficiency virus and ccr5 gene transcripts by means of rna-interference
SUBSTANCE: what is described in a cartridge genetic maker based on GeneClip-Ul-neo vector producing three siRNA - reproduction inhibitors of human immunodeficiency virus type 1 and human CCR5 gene. The maker contains a fragment of the length 27 bp related to a conservative region of a HIV-1 genome reverse transcriptase domain, a fragment of the length 27 bp related to the other conservative region of the HIV-1 genome reverse transcriptase domain, as well as a fragment 19 bp related to CCR5 gene mRNA. The invention can be used in medicine and scientific research.
EFFECT: invention allows producing more effective anti-HIV preparations based on interfering RNA produced in cells by means of the introduced cartridge genetic maker containing palindromes for three siRNA production.
3 dwg, 2 tbl, 2 ex
The invention relates to the fields of medical and molecular genetics, and gene therapy, and is associated with the previously identified targets for RNA interference in human immunodeficiency virus type 1 (HIV-1) to create a genetic cassette design. This cassette expresses two biologically active siPHK that leads to the attack of the relevant targets in the transcripts of the virus, and potent inhibition of its reproduction in human cells. The specified cassette genetic design Express hairpin RNAS that are processed in vivo to the biologically active siPHK. Last effectively delivered in protein complexes responsible for cleavage of transcripts of the virus, and this is achieved by the inhibition of its reproduction. In addition, the cassette expresses and new biologically active siPHK attacking mRNA gene CCR5 person. Specified gene encodes a co-receptor, facilitating penetration of the virus in T-lymphocytes of human rights. This genetic construct can be used in medicine for gene therapy of AIDS and HIV infection, as well as in scientific research for powerful anti-HIV products.
Currently, the main approach for the treatment of AIDS and HIV infection is a prescription antiviral chemotherapy, including drugs that act on key fermentive-1 - reverse transcriptase, integrase, and protease. However, despite the large number of drugs, there is the issue of the effectiveness of antiviral therapy. The main reason for treatment failure - adaptive mutagenesis of the virus, leading to the emergence of variants of viruses resistant to antiviral drugs. Serious problems are also toxicity and high cost of used medicines.
It is known the use of ribozymes and antisense RNA for the treatment of HIV infection [Dorman N. And Lever, A.M. (2001) RNA-based gene therapy for HIV infection. HIV Med., 2, 114-122; Michienzi, A., Conti, L., Varano Century, Prislei S., Gessani S., and Bozzoni I. (1998) Inhibition of human immunodeficiency virus type 1 replication by nuclear chimeric anti-HIVribozymes in a human T lymphoblastoid cell line. Hum. Gene Ther., 9., 621-628; Veres G., Junker, U., Baker J., Barske C, Kalfoglou C, lives H., S. Escaich, Kaneshima H., and Bohnlein, E. (1998) Comparative analisis of intracellularly expressed antisense RNAs as inhibitors of human immunodeficiency virus type 1 replication. J. Virol., 72, 1894-1901.], which block multiple HIV genes, which reduces its activity.
The closest to this invention are genetic constructs that give expression siPHK within HIV-infected cells (RF Patent 2324738, "Genetic design based on the vector plasmid pEGFP-Nl, producing siPHK inhibitors reproduction of human immunodeficiency virus type 1 (options)".
The present image is the buy differs in the present genetic structure expressed not one but three siPHK, two of which focused on two other targets in the transcripts of the virus, and one targeted to the mRNA of the gene CCR5 human encoding a cellular co-receptor involved in the penetration of the virus in T-lymphocytes. In addition, in the present genetic structure uses a different initial vector.
The technical result of the invention is the use of two previously identified targets RNA intreference in the virus (patent of the Russian Federation dated 29.08.2008, registration number 2008134932) and one new targets CCR5 mRNA described in this Application, to obtain a higher effect of suppressing the virus. This cassette genetic structure contains three palindrome intended for the education of RNA hairpins". The expression of such constructs in human cells leads to the formation of three interfering RNA (siPHK). The proposed genetic structure encode biologically active siPHK, which were selected using non-viral test systems using cultures of human cells.
Presents the technical result is achieved by creating a genetic cassette design on the basis of the vector GeneClip™ U1 Neomycin (Ac. number AY745746), http://www.promega.com/pnotes/89/12416_21/12416_21 .pdf.
Created genetic structure length 195 bp contain stawk the DNA specified in Table 1:
Table 1 shows 19-27-nucleotide sequence corresponding to the transcripts of the virus or CCR5 mRNA. Lowercase letters in bold in the following order specified regions corresponding to the semantic chain mRNA virus (2435..2461), antisense chain; semantic chain mRNA virus (2304..2330), antisense chain; semantic chain mRNA gene CCR5 (47..65), antisense chain. The DNA sequence presented in orientation 5'-3'. Lowercase letters shown texts loops and spacers containing artificial sites for restricted EcoRI or BamHI (bold lowercase letters). Loops are formed in the transcript after annealing of complementary areas of palindromes. The texts of genetic constructs correspond to the strain of HIV-1 subtype A (HIV-1 subtype A), the number of sequences in GenBank - AF316544. The numbering of the CCR5 gene region is specified by a sequence of GenBank - U54994. This design is intended for defeat of two regions of mRNA that encode conservative region of the reverse transcriptase (RT) virus, and the 5' region of the mRNA of the gene CCR5 person in the first codons.
Specified genetic structure was tested using non-viral systems. This system is intended for quantitative evaluation of biological activity siPHK, which are formed in vivo by expression of genetices the x structures. It was created on the basis of the vector psiCHECK™-2 (Promega, number sequences in GenBank AY535007). The vector contains a gene of the Firefly luciferase gene and the luciferase coral (Renilla). These luciferase cause the illumination of different wavelengths and their expression can be measured using a luminometer. 3' end non-coding region of the gene Renilla inserted a short region of the viral genome (domains)corresponding to the selected target RNA interference in the transcripts of the virus. The domains were obtained using chemical synthesis of DNA oligonucleotides. Table 2 presents the texts of the domains that cloned in the vector psiCHECK™-2 sites restricts XhoI and NotI.
In table 2 lowercase letters shows the region containing the sites of the restriction endonucleases XhoI and NotI, and two spacer between fragments of the genome of HIV-1 and CCR5 cDNA. Bold font shows the region of the viral genome and cDNA CCR5 length of 28 bp each, which correspond to the target RNA interference. The numbering of the areas are listed according to the sequence of the virus (AF316544) and CCR5 cDNA (U54994) in the following order: a target for anti-HIV-1, anti-HIV-2, anti-CCR5-8.
Testing the biological activity of the cassette is carried out in experiments on co-transfection culture of human cells. Using two DNA preparation: cassette genetic constructs containing the promoter U1 RNA polymerase I coding coding three siPHK, as well as a DNA plasmid-based vector psi-CHECK-2, containing three cloned domain relevant data siPHK (target RNAi) (table 2). In human cells at the genetic structures is expressed paste containing palindromes. Synthesized RNA forms a hairpin, which is a natural substrate of Disera, enzyme, the clip 21 nucleotide duplexes RNA from studs. If Dicer working on the duplex RNA longer than 19 base pairs (natural substrate), then he, being associated with double-stranded siPHK, actively participate in its "loading" in the protein complex RISC. In this complex there is an "unwinding" of the circuits siPHK and freeing it from one of them (the so-called "passenger"-chain). If the complex remains antisense siPHK, it is used as a "gunner", recognizing the complementary target transcripts cells. After binding of the complex with the corresponding mRNA is cutting one of its proteins, which leads to shut down expression of the corresponding gene of the virus and violates its reproduction in the cell host. In transfected cells Renilla mRNA containing a 3'-non-coding region of the test target RNA interference, is cut, which leads to a sharp decrease in the blue glow, caused by the luciferase (478 nm). Yellow-green is e glow, called luciferase Firefly Photinus pyralis (557 nm), encoded by the same DNA psiCHECK-2, serves as an internal control.
This study used ten criteria for selecting the sequence of the target RNA interference of mRNA of a gene CCR5. They are important to ensure that the antisense chain siPHK remained in the RISC complex. Each are given in Table 1 inserts in the genetic structures corresponds to at least ten of the eleven following criteria were used to select crustal 19-sequence polymorphisms siPHK:
composition G/C 30-52%
- at least 3 A/U in position 15-19 semantic chain
- lack of internal repeats
- And at position 19 of the sense circuit
And in position 3, the semantic chain
- U at position 10 of the sense circuit
- no G/C at position 19 of the sense circuit
- the absence of G at position 13 of the sense circuit
- the absence of homopolymer tracts (A)4-5or (T)4-5in the semantic chain
- localization of a target in a conservative region of the viral genome
- lack of homology to known transcripts in the human genome.
Part of the above selection criteria biologically active siPHK published [Reynolds, A., Leake d, Boese q, Scaringe s, Marshall W.S., Khvorova, A. (2004) Rational siRNA design for RNA interference. Nature Biotechnol. 22, 326-330] and presented on the website http://www.dharmacon.com.
See the table 1 genetic cassette design to deliver a targeted effect on the mRNA of the virus in human cells using siPHK, containing substances that occur in cells and CCR5 mRNA. therapeutic result of this effect is the suppression of the reproduction of HIV-1 in human cells.
For human immunodeficiency virus the first type is characterized by a rapid emergence and selection of mutant variants with resistance to various anti-viral drugs. To solve viral variability obtained genetic cassette design, attacking different targets and therefore largely independent of the variability of the virus.
Figure 1 shows a physical map of the vector GeneClipU1-neo.
Figure 2 shows the physical map of the vector psiCHECK-2.
Figure 3 presents the results of examining the effectiveness of RNA interference caused created genetic cassette design on non-viral test the system.
Example 1. Description of the genetic structure and method of reception
Vector GeneClipUl-neo registered in GenBank (Accession # AC AY745746) has a size of 4,758 kb (kilobases - thousand base pairs), and is characterized in that it contains:
the start site of RNA polymerase T7: 1
U1 premotor RNA polymerase II person: 46-438
10 bp spacer: 439-448
U1 terminator: 449-465
the promoter of RNA polymerase SP6: 527-546
early enhancer/promoter of the SV40 virus: 798-1216
signal the beginning of replication of SV40 virus: 1114-1179
gene neo: 1251-2045
synthetic poly(A): 20802128
the gene of β-lactamase: 3080-3940
the promoter of RNA polymerase T7: 4742-3.
In this vector, which comes Promega in linear form, insert a chemically synthesized DNA oligonucleotides, which correspond to the genetic structures that produce hairpin RNA (table 1).
Vector psiCHECK-2 is registered in GenBank (Accession # AY535007), has a size 6,27 kb and is characterized in that it contains:
early enhancer/promoter of the SV40 virus: 7-425
chimeric intron: 489-621
the promoter of RNA polymerase T7: 666-684
gene luciferase Relilla: 694-1629
synthetic poly(A): 1688-1736
the promoter HSK-TK: 1744-2496
gene luciferase Firefly: 2532-4184
late signal poly(A) SV40: 4219-4440
the gene of β-lactamase: 4587-5447.
In this vector in the sites XhoI and NotI of polylinker insert chemically synthesized DNA oligonucleotides that correspond to the selected target RNA interference (table 2).
Example 2. Assessment of the effectiveness of RNA interference caused created by genetic structures in non-viral system
On the eve of the co-transfection (18-20 hours) cells NEC scatter in the wells of a 24-hole tablet Nunc 50 testlets in 500 μl of culture medium DMEM (Paneco)containing glutamine and serum (complete medium), per well. On the day of the experiment, prepare DNA samples for co-transfection at room temperature following the way (one experimental point). 200 μl of DMEM medium containing no serum (SFM), add 100 ng DNA genetic structure in the vector GeneClip-U1-neo (see Table 1) and 10 ng DNA plasmids containing the corresponding cloned domain vector psiCHECK-2 (see Table 2). After mixing, add 1 ál viersprong reagent TransFast (Promega), shake the mixture and incubated for her 15 minutes Then sucked off the medium from the wells with those cells, shake the tube with a mixture of DNA TransFast and immediately add the mixture to the wells with cells. After incubation for 1 hour at 37°C in CO2-incubator added to the wells in 600 μl of medium with serum, mix and cuberoot tablet 48-72 hours.
To measure the expression of luciferase medium over the cells in the wells sucked off, add 100 ál of a solution of trypsin-EDTA (Paneco) per well, after 10 min incubation at 37°C in CO2-incubator add 100 ál of complete medium, carry a fully-cell suspension in a test tube, Eppendorf 0.5 ml, precipitated by centrifugation at room temperature in an Eppendorf centrifuge (5 min 2000 rpm), the supernatant is sucked off, suspended cells in 70 μl of 1xPBS and conduct the measurement light luciferase Firefly and Renilla according to the guidelines set Dual-Luciferase Reporter Assay System (Promega) on a luminometer Reporter Microplate Luminometer (Turner BioSystems). Figure 3 shows the results of testing of genetic constructs on neverenoughtime. It is seen that the genetic constructs specifically inhibit expression of the target for 60-96%.
Cassette genetic design of anti-HIV-1-anti-HIV-2-anti-CCR5-8-based vector GeneClip-U1-neo containing the insert in the area of single-stranded ends, indicated in figure 1, producing three siPHK inhibitors reproduction of human immunodeficiency virus type 1 and gene expression of human CCR5 and contain two fragments of the genome of HIV-1 long 27 bp each, corresponding to two conservative areas of the domain of the reverse transcriptase of the genome of HIV-1 (region 2435 virus..2461 shown in the first line, and the region 2304..2330 - second line), and 19 bp region corresponding to the gene CCR5 (47..65, shown in the third line):
where lower case letters shows the area of the loops between the parts of palindromes that can form double-stranded duplexes.
SUBSTANCE: recovered polynucleotide coding aminopeptidase containing a nucleotide sequence presented in the description is presented. Also, polynucleotide hybridised with said polynucleotide in the high stiffness environment is presented. An expression vector containing said polynucleotide, and an applicable host cell are described. Polypeptide related to the sequence presented in the description and being aminopeptidase is presented. A method of producing said polypeptide involving the stages of applicable host cell transformation by said polynucleotide or vector, cell cultivations in the medium adequate for said polynucleotide expression, and optional polypeptide purification from said cell or culture medium is offered. Besides, there has been described diagnostic technique for Aspergillus-infection in an organism involving the stages: a) recovery of a biological sample from said organism which is supposed to be Aspergillus infected, b) recovery of nucleic acid from said sample, c) determination whether said recovered nucleic acid contains polynucleotides hybridised with polynucleotide recovered from Aspergillus niger.
EFFECT: invention allows diagnosing Aspergillus-infection in the organism.
13 cl, 3 tbl, 11 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to biotechnology and immunology. Versions of a human CD37 specific antibody are presented, each containing a variable site of a light and heavy chain. A coding nucleic acid and a based expression vector are described. There are disclosed: a host cell containing a vector, a method of producing the antibody with using the cell, and also a composition for decreasing the B-cell count and a method of treating diseases associated with aberrant B-cell activity, with using the CD37-specific antibodies.
EFFECT: use of the invention provides specific loss of 80% BJAB cells at the antibody concentration 10 mcg/ml, and also increases the survival rate of Daudi mice as compared with Rhithuximab therapy that can find application for treating various tumours.
39 cl, 39 dwg, 7 tbl, 18 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention relates to field of biotechnology and deals with conjugates "caliheamicin derivative-carrier". Essence of invention includes methods of obtaining conjugates "monomer cytotoxic medication-caliheamicin/carrier" with considerably higher loading with medication than in earlier described methods, with low aggregation degree and with low content of low-conjugated fraction (LCF), as well as conjugates "cytotoxic medication-caliheamicin derivative/anti-CD22-antibody". Invention also includes compositions, which contain conjugate with CD22 antibody and application of said conjugates.
EFFECT: invention makes it possible to create conjugates with high loading of cytotoxic medication.
181 cl, 11 ex, 13 tbl, 29 dwg
SUBSTANCE: described is a polypeptide, having phytase activity, selected from: a polypeptide with an amino acid sequence given in the description of the invention, a fragment of that polypeptide, and a polypeptide which exhibits at least 90% identity with the said polypeptide. Described also is a polynucleotide which codes the said polypeptide. An expression cassette and an expression vector which contain the described polynucleotides are disclosed. A host organism producing the described polypeptide is disclosed. Furthermore, a feed additive and animal feed containing the said polypeptide, host organism or metabolites thereof is obtained. The invention discloses use of the said polypeptide to prepare a feed additive or animal feed and for hydrolysis of myoinositol hexakisphosphate to inorganic monophosphate, to myoinositol with lower degree of phosphorylation and to free myoinositol.
EFFECT: invention enables to obtain inorganic monophosphate, myoinositol with lower degree of phosphorylation or free myoinositol from a complex compound of myoinositol hexakisphosphate and vary the ration of domestic animals.
14 cl, 11 dwg, 9 tbl, 2 ex
SUBSTANCE: offered are versions of antibodies each of which is specifically bound with IGF-IR, inhibits its activity and is its antagonist, not exhibiting substantially IGF-IR agonist activity. Each of the antibodies is characterised at least by the presence of a variable area of a heavy and easy chain. There are described: antibody conjugates with cytotoxic agents, and also versions of a pharmaceutical composition for cancer diagnosing and therapy, methods of cancer treatment and diagnosing, a cancer diagnosing reagent - on the basis of the antibodies. There are disclosed: a method of producing the antibodies; nucleic acid (NA) coding the antibody; an expression vector containing NA. Offered is a hybridoma EM 164 producing the antibody under the invention, deposited in ATCC, No. PTA-4457, and also the use of the antibody for IGF-IR linkage. The use of the invention presents the antibodies which allow inhibiting MCF cell growth approximately in 12 times that is higher approximately in 5 times than hen using the antibody IR3, and can be used for cancer diagnosing and treatment expressing higher levels of receptor IGF-I, such as breast cancer, colon cancer, lung cancer, prostate cancer, ovarian cancer, synovial carcinoma and pancreatic cancer.
EFFECT: more efficient diagnosing and treatment of said cancers.
58 cl, 28 dwg, 10 tbl, 1 ex
SUBSTANCE: recombinant technique is used to produce proteins consisting of the module SEQ ID NO: 3, involving replacement of serine and/or glutamate with lysine and/or cysteine and/or addition of the amino-terminal TAG according to SEQ ID NO:20-24 or carboxy-terminal TAG according to SEQ ID NO:25-28, which contain lysine or cysteine along with other amino acids.
EFFECT: invention enables to obtain modified spider silk protein, capable of binding with other substances through a lysine or cysteine residue with preservation of structural integrity of the spider silk protein.
21 cl, 8 dwg, 2 tbl, 1 ex
SUBSTANCE: invention can be used for determining a functional status of primary signalling pathways of a cell. Substance of the invention are lentiviral reporter vector structures containing reporter markers. The invention allows predicting the tumour sensitivity to therapeutic exposures and developing an individual therapeutic approach.
EFFECT: creating the lentiviral reporter vector structure optimal for determining the activity of transcriptional factors reflecting the functional status of signalling pathways of the cell.
6 cl, 16 dwg
SUBSTANCE: there is offered a method of engineering yeast strains - stable human growth hormone (somatotropin) producers. Also, there are offered two strains Y-3506 of Russian National Collection of Industrial Microorganisms and Y-3507 of Russian National Collection of Industrial Microorganisms - stable human somatotropin producers. The producer strains have been prepared by sequential integration of expression plasmid into a recipient stain genome. Each plasmid carries in its structure a somatotropin gene (GH1); fused with a leader sequence controlled by the GAL1 promotor, as well as one of the genes URA3, LEU2, TRP1 and HIS3 complementing auxotrophic recipient strain mutations.
EFFECT: efficacy of the produced strains is 100-130 mg of somatotropin per 1 l of the medium.
3 cl, 7 dwg, 6 ex
SUBSTANCE: elaborated is method of obtaining factor, which takes part in process of control of appetite and/or body weight. Also described are genes, obtained by said method, polypeptides, coded by said genes, intended for treatment, control or diagnostics of diseases, associated with eating disorders and/or control of body weight. Invention also relates to substances, which inhibit activity of said genes or said polypeptides, intended for treatment, control or diagnostics of diseases, associated with process of appetite and/or body weight control.
EFFECT: using tiasolidindions, possessing PPARγ-agonistic activity, it is possible to obtain genes and polypeptides, involved into regulation of appetite and/or body weight reduction.
27 cl, 41 dwg, 35 ex
SUBSTANCE: recombinant plasmid pETPHOFus enabling synthesis of an anthrax lethal factor substratum as a part of one polypeptide with a mutated reporter enzyme of alkaline phosphatase Escherichia coli. Said fused polypeptide represents a new high-sensitivity anthrax lethal factor substratum. Besides, an Escherichia coli BL-PHOFus strain being a producer of the fused polypeptide is offered. The strain provides a no-fermentation high-yield synthesized protein not less than 30 mg in 1 litre of a liquid culture. The prepared anthrax lethal factor substratum shows hypersensitiveness and high specificity to LF splitting and is able to detect proteolytic activity of the anthrax lethal factor in concentration up to 1 pM.
EFFECT: invention can find application in clinical recognition of anthrax disease; for detecting hot spots of anthrax, for screening the compound libraries and searching anthrax lethal factor activity inhibitors.
2 cl, 4 dwg, 4 ex
SUBSTANCE: new 02_AG.RU.09RU2204 1 type human immunodeficiency virus strain, recombinant subtype 02AG is recovered in the territory of Russia in a 18-year-old sexually infected woman with acute HIV-infection diagnosed. The strain is supposed to be included in the national strain panel, also can be used for studying of the efficacy of therapeutic and preventive chemotherapeutic and vaccine preparations, improving the diagnostic techniques for HIV-infection. The HIV-1 02_AG.RU.09RU2204 human immunodeficiency virus strain is deposited in the Collection of Microorganisms of Federal State Research Institution State Science Centre Vector of Federal Service for Supervision of Consumer Rights and Human Welfare.
EFFECT: improved diagnostic accuracy.
4 dwg, 4 ex
SUBSTANCE: composition contains HIV fusion protein. Also, there is disclosed the application of such composition for manufacturing a drug for HIV immune response elicitation and a method for immune response elicitation with using thereof.
EFFECT: HIV immune response elicitation.
31 cl, 8 dwg
SUBSTANCE: invention relates to field of biotechnology, virology and medicine. Described is immunogenic composition of preliminarily determined inactivated strains of human immunodeficiency virus (HIV). Inactivation is carried out by means of psoralen and ultrasonic radiation. Composition is made more efficient by removal of structural HIV peculiarities, which prevent immune response. In particular, sialic acid is removed in order to enhance immune recognition of composition and for weakening of complimentary H factor binding. Also for prevention of complimentary H factor binding CD55 and CD59 are removed. Determination of strains for inactivation can be realised by means of immunotherapeutic genotyping or probabilistic assessment of infection risk.
EFFECT: invention can be used in medicine.
30 cl, 3 dwg
FIELD: chemistry; biochemistry.
SUBSTANCE: invention relates to biotechnology, virology and medicine. Vaccines can be used to treat individuals chronically infected with HIV by inducing a protective cell-mediated response in these individuals against the same HIV subtype which was used to make the vaccine. Invention can be used in medicine.
EFFECT: obtaining an intact human immunodeficiency virus of a defined subtype which can be used in making vaccines and pharmaceutical compositions containing such vaccines.
21 cl, 2 dwg, 4 ex
FIELD: chemistry; biochemistry.
SUBSTANCE: present invention relates to biochemistry and specifically to a modified polypeptide of a HIV-1 gp41 envelope glycoprotein, a polynucleotide which codes the modified polypeptide and an expression vector which contains a coding modified polypeptide of the HIV-1 gp41 envelope glycoprotein. The modified polypeptide of the HIV-1 gp41 envelope glycoprotein contains an amino acid sequence SEQ ID NO: 1 or SEQ ID NO: 14, where the sequence between positions 603 and 615 or 598 and 622 SEQ ID NO: 1 or the sequence between positions 530 and 542 or 525 and 549 SEQ ID NO: 14 is replaced by a linker fragment which is an oligopeptide SEQ ID NO: 2.
EFFECT: improved solubility of a modified polypeptide of the HIV-1 gp41 envelope glycoprotein without changing its immunogenic reactivity.
10 cl, 8 dwg, 1 tbl, 3 ex
FIELD: medicine; veterinary science.
SUBSTANCE: vaccine contains mixed avirulent purified antigens of strain No.101 "ВНИИ3Ж-ДЕП" of cattle rotavirus and of strain "ВНИИ3Ж-ДЕП" of cattle coronavirus in amount to provide protective immunoactivity in an animal's organism when a desired preparation is introduced, and an oil adjuvant in effective ratio. In immunised animals, the vaccine induces high level of antibodies against cattle rota- and protects the cattle stock of sex-age groups from the diseases as stated above. The invention can be used in veterinary science.
EFFECT: new compounds possess effective biological activity.
14 cl, 7 tbl, 3 ex
SUBSTANCE: synthetic peptide with the sequence (III), being a derivative of glycoprotein gp41 of a HIV-1 virus also is suitable for application in immunologic analyses for detection of the infections caused by a HIV-1 virus. The antigenic composition, the method of obtaining of a peptide, the method of detection in vitro antibodies against a HIV-1 and a set for its realisation are besides, offered.
EFFECT: rising of the efficiency of the method.
9 cl, 4 tbl, 3 ex
SUBSTANCE: vaccine contains: purified avirulent antigen material from cattle coronavirus strain "ВНИИЗЖ", produced in sensitive biological system; special adjuvant and saponin additives. The strain is deposited in Federal State Institution (FSI) "ВГНКИ" collection under reference number: cattle coronavirus strain culture "ВНИИЗЖ-ДЕП".
EFFECT: vaccine has high immunogenicity; it is innocuous, doesn't cause reaction; effectively protects cattle livestock against epizootic coronavirus, circulating around Russian territory.
22 cl, 8 tbl, 7 ex
SUBSTANCE: offered method provides introduction to mammal of lymphocyte conditioned medium containing "untrained" T-cells cultivated with CD3/CD28 antibodies covered granules, in combination with antigen. Method can be used in medicine.
EFFECT: allows for enhanced patient's immune system response to vaccine or another immunotherapeutic agent.
10 cl, 37 dwg, 3 tbl, 1 ex
FIELD: medicine, polymers.
SUBSTANCE: invention relates to conjugates consisting of a water-soluble polymer of molecular mass from 200 to 20000 Da and representing polyethylene glycol or alkyl chain to which two molecules of synthetic peptides, not less, are bound by reactive functional group and wherein each peptide comprises amino acid sequence originating from region HR1 or HR2 of human immunodeficiency virus (HIV) gp41. Invention relates to methods for using these conjugates for delivery inhibition of to HIV target-cell by addition of indicated conjugates in the amount providing effective inhibition of cell infection with indicated virus. Also, invention relates to methods for preparing conjugates by functional adding of each molecule of synthetic peptide to polymer through reactive functional group.
EFFECT: valuable biological properties of conjugates.
27 cl, 2 dwg, 6 tbl, 6 ex
FIELD: biotechnology, genetic engineering, molecular biology.
SUBSTANCE: invention proposes recombinant DNA pcDNA-TCI that has molecular mass 4.3 x 103 kDa and size 6 657 nucleotide pairs. The proposed recombinant plasmid DNA provides expression of artificial gene TCI comprising cytotoxic T-cellular epitopes of HIV-1 in eucaryotic cells. Also, invention proposes recombinant attenuated strain of bacterium Salmonella enteritidis E-23 pcDNA-TCI. The proposed strain is obtained by genetic transformation of the strain Salmonella enteritidis E-23 with plasmid pcDNA-TCI. The strain provides delivery DNA-vaccine in eucaryotic cells as the recombinant plasmid DNA pcDNA-TCI. Proposed groups of inventions provide inducing the expressed specific humoral and cellular immune response to HIV-1 in body. Proposed group of inventions can be used in medicine and virology for construction of live DNA-vaccine against HIV-1.
EFFECT: valuable biological and medicinal properties of strain and vaccine.
2 cl, 3 dwg, 3 tbl, 5 ex