Method of amplification of immune response to antigene of mammal
SUBSTANCE: imiquimod or resiquimod is injected locally to a mammal over 12-26 hours after injection of nucleotide sequences encoding granulocyte-macrophage colony-stimulating factor and antigen peptide or protein.
EFFECT: significant amplification of immune response of mammal to antigen.
18 cl, 23 dwg, 1 tbl, 1 ex
The text descriptions are given in facsimile form.
1. Method of enhancing the immune response of a mammal to an antigen, including the introduction of the following components:
(1) the agonist of TLR-7, selected from imiquimod or resiquimod;
(2) the nucleotide sequence encoding granulocyte-macrophage colony-stimulating factor; and
(3) the nucleotide sequence encoding the antigenic peptide or protein,
where the component (1) is administered locally through 12-26 h after components (2) and (3).
2. The method according to claim 1, where one or more than one nucleotide sequence is a DNA sequence.
3. The method according to claim 1, where the nucleotide sequence is encoded in a DNA plasmid.
4. The method according to claim 1, where the nucleotide sequence (3) encodes a protein P501S or its derivative, which can enhance the immune response in vivo, and this immune response able to recognize tumor cells or tumor expressing P501S.
5. The method according to claim 1, where the nucleotide sequence (3) encodes a protein MUC-1 (mucin-1) or its derivative, which can enhance the immune response in vivo, and this immune response able to recognize tumor cells or tumor expressing MUC-1.
6. The method according to claim 5, where the protein MUC-1 or its derivative does not contain any duplicate elements (perfect or imperfect).
7. The method according to claim 5, g the e protein, MUC-1, or its derivative does not contain any committed repeated elements.
8. The method according to claim 5, where the protein MUC-1 or its derivative contains from 1 to 15 repetitive elements.
9. The method according to claim 5, where the protein MUC-1 or its derivative contains 7 committed repeated elements.
10. The method according to claim 5, where the nucleotide sequence encoding a protein, MUC-1, or its derivative, has modified codons.
11. The method according to claim 5, where the nucleotide sequence encoding a plot with imperfect repeat, is the value of the relative usage of synonymous codons at least 0.6.
12. The method according to claim 5, where the nucleotide sequence encoding an imperfect duplicate items protein MUC-1 or its derivative has a level of identity with MUC-1 wild type DNA in the appropriate non-recurring areas less than 85%.
13. The method according to claim 5, where the protein MUC-1 or its derivative has a modified repeat items (VNTR (variable number of tandem repeats)), for example, mutants with reduced glycosylation.
14. The method according to claim 5, where the protein MUC-1 or its derivative is a protein or anywhereman alien T-cell epitopes.
15. The method according to 14, where protein MUC-1 or its derivative is a protein or anywhereman with P2, or RSO, or their fragments.
16. The method according to 14, where alien T-cell epitopes introduced into the molecule or on any the second end of the protein, MUC-1, or its derivative.
17. The method according to any one of claims 1 to 16, where at least one media component (2) or (3) is a Golden grains and suitable for delivery of drugs through the particles.
18. The method according to 17, where the media components (2) and (3) is a Golden grains, and the component (1) prepared for sequential injection.
SUBSTANCE: invention relates to field of biotechnology, namely, to obtaining genetically modified cell lines and can be applied in medicine for immunotherapy and immuno-prophylaxis in patients with malignant neoplasms. By means of recombinant method line of cells of human melanoma KG is obtained, which secretes recombinant granulocytic-macrofagal colony-stimulating human factor. Obtained line is deposited with Specialised cell culture collection of vertebrates of Russian cell culture collection under number RCCC ("П") 699"Д".
EFFECT: line of human melanoma cells KG possesses stable cultural, morphological and immunological characteristics and possesses ability to secrete recombinant human GM-CSF, remaining after cell inactivation with ionising irradiation.
FIELD: gene engineering.
SUBSTANCE: invention can be used for production of recombinant polypeptide of human granulocyte colony-stimulating factor. Recombinant plasmid DNA is constructed in vitro. It includes synthetic gene of human granulocyte colony-stimulating factor, strong constitutive promoter A3 from the early stage of bacteriophage T7 and synthetic section - translation enhancer (TREN) of gene 10 in bacteriophage T7. This DNA in combination with high copy number of plasmid and optimisation of cultivation conditions ensures constitutive biosynthesis of target protein in transformed by this DNA race of Escherichia coli SGK25/pA3GF with high yield.
EFFECT: constitutive biosynthesis of target protein in cells; high yield.
2 cl, 4 dwg, 7 ex
FIELD: medicine, endocrinology, pharmacy.
SUBSTANCE: invention proposes an agent eliciting antidiabetic activity. Agent represents nonglycosylated recombinant human granulocyte colony-stimulating factor. It differs from native preparation by absence of glycosyl group and the presence of additional methionine residue by its N-end. Effect of agent is associated with mobilization and migration of bone marrow mesenchymal stem cells, homing into pancreas. This results to reparation of insulin-producing activity of organ and normalization of the peripheral blood glucose level after monotherapy with recombinant human granulocyte colony-stimulating factor.
EFFECT: valuable medicinal property of agent.
3 tbl, 2 dwg, 1 ex
FIELD: molecular biology.
SUBSTANCE: invention relates to isolated DNA fragment encoding horse GM-CSF, and isolated horse GM-CSF protein. Also disclosed are vectors and various compositions containing thereof. GM-CSF is useful as adjuvant for horse vaccination as well as non-specific immunity stimulator in veterinary.
EFFECT: new compositions for gorse vaccination.
18 cl, 2 dwg, 1 tbl, 7 ex
FIELD: biotechnology, microbiology, genetic engineering.
SUBSTANCE: invention describes construction of recombinant plasmid DNA pFGM17 encoding constitutive synthesis of polypeptide of human granulocytic-macrophagal colony-stimulating factor (GM-CSF) and consisting of Kpn I/Eco RI-fragment of plasmid pSPF1 DNA and artificial DNA sequence encoding signal peptide of the protein Caf1 from Yersinia pestis, and also Kpn I/Eco RI-fragment of intermediate plasmid pSK-GM comprising the synthetic human gene GM-CSF. Escherichia coli cells are transformed with plasmid DNA pFGM17 and strain E. coli BL21(DE3)/pFGM17 is prepared that is a producer of human polypeptide GM-CSF. Invention provides enhancing technological effectiveness and economy of process for preparing recombinant FM-CSF due to excluding the induction stage in biosynthesis process and in simultaneous increasing the yield of the end product by 2 times. Invention can be used for preparing human granulocytic-macrophagal colony-stimulating factor.
EFFECT: valuable properties of plasmid DNA and microorganism strain.
2 cl, 4 dwg, 4 ex
FIELD: genetic engineering, in particular production of human granulocyte colony-stimulating factor.
SUBSTANCE: Recombinant plasmid DNA pES3-7 with molecular weight of 3.63 MDa (5907 b.p.) is constructed. Said DNA consists DNA Ndel/Notl-fragment containing sequence of recombinant G-CSF artificial gene, β-lactamase gene; and plasmid pET22b(+) DNA Ndel/Notl-fragment containing promoter and terminator of T-RNA-polymerase transcription, amplifier of 17 phage 10 gene translation. Plasmid pES3-7 contains as genetic marker β-lactamase gene which determines resistance of E.coli cells transformed with plasmid pES3-7 to ampicillin, and unique restriction endonuclease recognition sites existing on the next distance to the right from Ndel-site: Xbal - 38 b.p.; Hpal - 1332 b.p.; Pstl - 4065 b.p.; Pvul - 4190 b.p.; Xhol - 5363 b.p. Obtained plasmid is used in transformation of Escherichia coli cells to produce strain E.coli BL21(DE3)/pES3-7 as subproducer of recombinant G-CSF. Method of present invention makes in possible to produce recombinant G-CSF with high yield (20-30 % based on total cell protein content).
EFFECT: simplified method for production of recombinant G-CSF with high yield.
2 cl, 2 dwg, 2 ex
FIELD: chemistry; medicine.
SUBSTANCE: claimed are polypeptide and respective polynucleotide zcytor17lig and molecules of antibody against human zcytor17. Human zcytor17lig is novel cytokine. Claimed invention also relates to methods of protein obtaining, its application for stimulation of immune reaction in mammal. Described is method of obtaining antibodies to said protein and respective antibodies.
EFFECT: polypeptides can be used in realisation of methods stimulation of immune system, proliferation and development of hemopoietic cells in vitro and in vivo.
17 cl, 3 dwg, 21 tbl, 47 ex
FIELD: chemistry; medicine.
SUBSTANCE: invention can be used in production of preparations for treatment of pre-diabetic states, type 2 diabetes and glucose tolerance disturbances. Novel peptides, demonstrating properties of selective receptor VPAC2 agonists, in particular ability to stimulate insulin synthesis and its release from β-cells of pancreas by glucose-depending method, as well as following reduction of glucose level in plasma.
EFFECT: increased efficiency of action and stability in comparison with natural peptides, possibility of their successful application in treatment of diseases.
14 cl, 18 dwg, 1 tbl, 10 ex
SUBSTANCE: present invention relates to biotechnology. Description is given of a single-strand T-cell receptor (scTCR), containing an α segment, formed by a sequence of a variable region in a TCR chain, joined with the N end of the extracellular sequence with constant region in the TCR chain, a β segment, formed by a sequence of the variable region of the α TCR chain, joined with the N end of the extracellular sequence with constant region of the β TCR chain, and a linker sequence, joining the C end of the α segment with the N end of the β segment, or vice versa. Extracellular sequences of constant regions of α and β segments are joined by a disulphide bond. Extracellular sequences of constant regions can correspond to constant regions of α and β chains of native TCR, cut-off at their C ends such that, cysteic residues, which form the inter-chain native disulphide bond of the TCR, are excluded, or extracellular sequences of constant regions which are in the α and β segments, can correspond to constant regions of α and β chains of native TCR, in which cysteic residues, which form the native inter-chain disulphide bond, are replaced by another amino acid residue, or there is no uncoupled cysteic residue, which is in the β chain of the native TCR. This invention makes available a new class of alpha/beta analogues of scTCR, in which there is a disulphide bond between residues of a single amino acid, contributing to stability of the bond between the alpha and beta regions of the molecule.
EFFECT: such TCR are suitable for screening or for therapeutic purposes.
3 cl, 14 dwg, 3 ex
FIELD: medicine; biotechnology.
SUBSTANCE: present invention concerns biotechnology. The agent regulating a differentiation of cells-natural killers, containing in quality of an effective ingredient one or several genes is described. The genes used in the invention, are described in demand materials.
EFFECT: reception of the new agent regulating a differentiation of cells-natural killers.
6 cl, 4 dwg, 5 tbl, 7 ex
SUBSTANCE: adhesive protein containing protein and peptide repeated at least once, which is combined to carboxy- and/or amino-ends of the protein, is described. Polynucleotide containing nucleotide sequence, which encodes the adhesive protein, is represented. Expression vector containing functionally incorporated nucleotide sequence, which encodes the adhesive protein, is described. Transformant, which is transformed by the vector selected from the group, which contains prokaryote, eukaryote and cells, derived from eukaryotes, is represented. The methods of producing and purification of the adhesive protein are represented. Adhesive containing the adhesive protein as an active component in efficient number is offered. The method for regulation of adhesive strength of the present adhesive, including treatment with the substance selected from the group, which contains oxidising agent, filler and surfactant, or regulation of concentration of the adhesive protein, which is the surfactant of the adhesive, is described. Covering agent containing the adhesive protein as an active component in efficient number is offered.
EFFECT: present invention allows producing adhesive protein, which can be efficiently produced in volume to use them instead of chemical adhesive.
42 cl, 33 dwg, 1 tbl, 17 ex
FIELD: chemistry; biochemistry.
SUBSTANCE: present invention pertains to genetic engineering, more specifically to chimeric polypeptides, containing an antagonist of growth hormone receptor. The invention can be used in medicine. The binding domain of the growth hormone is modified by substituting glycine amino acid residue in position 120 and is further modified in site 1, where at least one amino acid residue is substituted, which increases affinity of the growth hormone to its binding domain on the growth hormone receptor. The amino acid residue is then conjugated with the ligand-binding domain of the growth hormone receptor, through a peptide linker.
EFFECT: obtaining a highly effective antagonist of the growth hormone receptor with longer half-life, reduced immunogenesity and nontoxicity, compared to known mutant forms.
35 cl, 16 dwg, 1 tbl
FIELD: chemistry, biochemistry.
SUBSTANCE: invention relates to area of gene engineering and biotechnology and can be used for labeling biological objects. The nucleic acid molecule was extracted that encoded the fluorescing protein selected from fluorescing proteins represented by the Copepoda Crustacea biological kinds and fluorescing mutants of the aforesaid proteins. The aforesaid nucleic acid was functionally bonded to appropriate elements of expression regulation to be used in the method of producing aforementioned fluorescing protein. On the basis of the aforesaid extracted nucleic acid cloning and expressing vectors are produced as well as the expressing cassette. The cell and a stable cellular lineage, containing the aforesaid expressing cassette produce the fluorescing protein. Note that the said fluorescing protein, the nucleic acid encoding the protein above and expressive genetic structures containing the said nucleic acid, are used incorporated with the set designed to label a biological molecule. The fluorescing protein is also used in the methods of labeling a biological molecule, a cell or a cellular organella.
EFFECT: expanded biological objects labeling means.
12 cl, 9 dwg, 1 tbl, 7 ex
FIELD: chemistry, biochemistry.
SUBSTANCE: invention relates to gene engineering and biotechnology and can be used in a pharmaceutical industry. Cleared and extracted polynucleotide encodes protein with functional activity of a glucose carrier and the GLUT4 protein amino acid lineage wherein valine in position 85 is replaced by methionine. The protein under consideration features biological activity in yeast strains S. cerevisiae deprived of functional carriers of glucose and Erg4 protein.
EFFECT: production of protein with functional activity of glucose carrier in yeasty expressing system.
5 cl, 3 tbl, 1 ex
SUBSTANCE: invention claims compositions which can include one or several mammary gland tumour proteins, their immunogenic parts or polynucleotides encoding such parts. Alternatively the therapeutic composition can include antigen-presenting cell expressing mammary gland tumour protein, or T-cell specific to cells expressing such protein. These compositions can be applied in prevention and treatment of such diseases as mammary gland cancer. Invention also claims diagnostic methods based on determination of mammary gland tumour protein or mRNA encoding such protein in sample.
EFFECT: use of peptides obtained from protein expressed from mammary gland by tumour in diagnostics and therapy of mammary gland cancer.
37 cl, 6 ex, 1 dwg
FIELD: medicine; biotechnologies.
SUBSTANCE: DNA-vaccine is intended for activation of the immune response against the cancer cells, containing the design of DNA operably coding survivin protein and cytokine CCL21, built in attenuated vector Salmonella typhimurium. The way of inhibition of growth of a tumour by vaccine introduction under the invention to a mammal is opened. The product containing the described vaccine is presented. The way of vaccination of a mammal against a cancer where the way includes a stage of introduction to a mammal of effective quantity of the described DNA-vaccine causing the immune response is offered.
EFFECT: expansion of assortment of DNA-vaccines causing the immune response and unexpectedly high level of cytotoxicity against tumoral cells.
21 cl, 42 dwg, 6 tbl, 22 ex
SUBSTANCE: invention concerns novel compounds of formula (1a), formula (1b), formula (1c) and formula (1d), as well as pharmaceutical composition based on them and their application in medicine obtainment. R1-R4, G, W, X, X1, U, V, a, b are defined in the invention claim.
EFFECT: compound with antagonistic effect on vasopressin V1A receptor.
73 cl, 133 ex