140 results about "Herpes simplex thymidine kinase" patented technology

A composition of matter comprising a vaccinia virus expression vector with a negative thymidine kinase phenotype and a negative vaccinia virus growth factor phenotype.

The present invention relates to compositions and methods for the treatment of tumor and / or malignant and / or cancerous cells. The present invention provides VSV vectors comprising nucleic acid encoding a cytokine, such as interleukin or interferon, or a suicide gene, such as thymidine kinase, or other biological protein, such as heat shock protein gp96, or endostatin or angiostatin, wherein said VSV vectors exhibit greater oncolytic activity against the tumor and / or malignant and / or cancerous cell than a wild-type VSV vector. The present invention also provides methods of making such vectors, host cells, expression systems, and compositions comprising such VSV vectors, and viral particles comprising such VSV vectors. The present invention also provides methods for producing oncolytic activity in a tumor and / or malignant and / or cancerous cell comprising contacting said cell with a VSV vector of the present invention. The present invention also provides methods for suppressing tumor growth comprising contacting said tumor with a VSV vector of the present invention. The present invention also provides methods for eliciting an immune response to a tumor cell in an individual.

Provided herein are constructs for genome editing or genetic engineering in fungi or protists, methods of using the constructs and media for use in selecting cells. The construct include a polynucleotide encoding a thymidine kinase operably connected to a promoter, suitably a constitutive promoter; a polynucleotide encoding an endonuclease operably connected to an inducible promoter; and a recognition site for the endonuclease. The constructs may also include selectable markers for use in selecting recombinations.

Provided herein are constructs for genome editing or genetic engineering in fungi or protists, methods of using the constructs and media for use in selecting cells. The construct include a polynucleotide encoding a thymidine kinase operably connected to a promoter, suitably a constitutive promoter; a polynucleotide encoding an endonuclease operably connected to an inducible promoter; and a recognition site for the endonuclease. The constructs may also include selectable markers for use in selecting recombinations.

A Mob− plasmid having a RSF1010 replicon, comprising a gene coding for Rep protein and said plasmid has been modified to inactivate gene related to mobilization ability. The present invention also describes a bacterium having an ability to produce useful metabolites, comprising the plasmid and said bacterium lack active thymidylate synthase coded by thyA gene and thymidine kinase coded by tdk gene, and a method for producing useful metabolites, such as native or recombinant proteins, enzymes, L-amino acids, nucleosides and nucleotides, vitamins, using the bacterium.

An anti-cytoplasm thymidine kinase-IgY antibody prepared from the C-terminal 31 peptide of human cervical cancer cell (HeLa)TK1, the application of the reagent kit prepared from said antibody in diagnosing tumor, the process for preparing said antibody and reagent kit, and its test method are disclosed.

Embodiments of the invention are directed methods that include a thymidine kinase deficient vaccinia virus. The methods include administering the vaccinia virus at increased viral concentrations. Further aspects of the invention include methods for inducing oncolysis or collapse of tumor vasculature in a subject having a tumor comprising administering to a subject at least 1×108 infectious viral particles of a TK-deficient, GM-CSF-expressing, replication-competent vaccinia virus vector sufficient to induce oncolysis of cells in the tumor.

This invention provides a recombinant adenovirus expression vector characterized by the partial or total deletion of the adenoviral protein IX DNA and having a gene encoding a foreign protein or a functional fragment or mutant thereof. Transformed host cells and a method of producing recombinant proteins and gene therapy also are included within the scope of this invention. Thus, for example, the adenoviral vector of this invention can contain a foreign gene for the expression of a protein effective in regulating the cell cycle, such as p53, Rb, or mitosin, or in inducing cell death, such as the conditional suicide gene thymidine dinase. (The latter must be used in conjunction with a thymidine kinase metabolite in order to be effective.)

The invention discloses attenuated, brightened and replication-controllable HSV recombinant virus, a preparation method and applications thereof. According to the present invention, thymidine kinase (TK) gene essential for replicating viruses in neurons and being a main virulence factor is knocked out by using a homologous recombination method, and subsequently a red or green fluorescent gene enhancement expression cassette is recombined into the genome of the virus to construct a series of novel recombinants viruses, wherein the toxicity is markedly low, the states of infected mice are good,the fluorescence signal is strong, and the expression of the recombinant viruses is limited at the injection site after the recombinant viruses are used in in-vivo animal center labeling; by combiningwith Cre-dependent AAV helper viruses capable of expressing TK in a compensated manner, the cell-specific transmonosynaptic loop tracing is achieved; and the recombinant HSV has wide application value in nervous system targeted gene transduction, neural network transsynaptic tracing, tumor disintegration, viral replication and pathogenesis mechanism, antiviral drug screening and other fields.

A method of treating ocular disorders (such as, for example, proliferative vitreoretinopathy or PVR) associated with replicating ocular cells by transfecting replicating ocular cells in vivo with a polynucleotide encoding an agent which is capable of providing for the inhibition, prevention, or destruction of the growth of the replicating ocular cells upon expression of the agent. The agent may be a viral thymidine kinase, and the polynucleotide encoding the agent may be contained in a retroviral vector. Once the replicating ocular cells are transduced with the retroviral vector, the patient is given a chemotherapeutic or interaction agent, such as ganciclovir, which kills the transfected replicating ocular cells.

The invention discloses a novel oncolytic virus based on vaccinia virus Tiantan strain. The thymidine kinase (TK) region of the virus contains a coding sequence of AIF-GM-CSF as shown in SEQ ID NO.1.The oncolytic vaccinia virus which can efficiently expressing a human AIF-GM-CSF gene is prepared by effectively combining the tumor suppression effect of gene therapy and the oncolytic effect of viral therapy. While the oncolytic virus of the vaccinia virus Tiantan strain achieves the oncolytic effect to pyrolyze tumor cells, human AIF is massively expressed to cause apoptosis of a great number of infected tumor cells; and a great number of human GM-CSF can be expressed, and NK cells or DC cells can be recruited into the inside of a tumor to kill the tumor or effectively represent a tumor antigen, so that proliferation and differentiation of cytotoxicity t cells can be improved, and multiple anti-tumor effects can be achieved. Compared with simple gene therapy or virus therapy, the malignant tumor killing performance of the novel oncolytic virus is reinforced.

A method of treatment of cancer, viral infections, and the like administers anti-TK1 antibody, constituted as the complete antibody or a fragment thereof. The antibody binds to the surface of cells expressing TK1 thereon. The antibody, with or without another agent bound thereto, may effect complement mediated lysis, antibody-dependent cell-mediated cell cytotoxicity, apoptosis, an immune response by the mammal, a reduction in cellular replication, a combination thereof, or the like for such cells. The antibody may be coupled to an immune response stimulator, a cytotoxin, an enzyme, a combination, or the like to effect the treatment desired.

The invention discloses a recombinant oncolytic vaccinia virus and a preparation method and application thereof. The thymidine kinase (TK) region of the virus comprises a coding sequence of a PD1 full-length antibody shown in the formula of SEQ ID NO.1. Through effective combination of the anti-tumor effect of gene therapy and the oncolytic effect of virus treatment, the oncolytic vaccinia virus for efficiently expressing the PD1 full-length antibody gene is prepared. When the oncolytic vaccinia virus produces the oncolytic effect of splitting tumor cells, the PD1 full-length antibody is efficiently expressed, the activity of PD1 on the surfaces of T cells is inhibited, the T cell immune response is activated and the dual antitumor effects are produced. Compared with the single gene therapy or viral therapy, the method using the recombinant oncolytic vaccinia virus improves the effects of killing malignant B cell lymphoma. Through virus replication-related gene deletion, the TK regionof the vaccinia virus genome is deleted so that specific replication of the viruses in the abnormally proliferating tumor cells is ensured and the viruses can not replicate in normal cells, so that the safety of the oncolytic vaccinia virus vector is greatly improved.

The present invention relates to new recombinant raccoon poxvirus vectors comprising two or more exogenous nucleic acid molecules, each encoding at least one feline protein, wherein at least two of the nucleic acid molecules are inserted into the hemagglutinin (ha) locus or the thymidine kinase (tk) locus, or at least one of the nucleic acid molecules is inserted into each of the hemagglutinin and thymidine kinase loci. Described herein are monovalent and polyvalent recombinant feline vaccines that encompass an immunologically effective amount of the recombinant raccoon poxvirus vectors and, optionally, a suitable carrier or diluent. The vaccine of this invention optionally includes additional feline antigens to provide broad spectrum protection to cats against a variety of feline pathogens. The invention further concerns the method for inducing a protective immune response to the feline pathogens in a cat by administering the recombinant vaccines.

The invention discloses a TK (Thymidine Kinase) gene removed recombinant VTT (Tian Tan strain) oncolytic VACA (Vaccinia Virus). A VTT oncolytic VACA is subjected to recombination transformation, a TKgene is removed, and an EGFP (Enhanced Green Fluorescent Protein) gene and a Luciferase gene are expressed. The invention further discloses a preparation method of the TK gene removed recombinant VTToncolytic VACA, which comprises the steps of: recombining a wild type VTT virus and a targeting plasmid aiming at the TK gene of the VTT virus in cells to obtain virus suspension; screening and purifying the virus suspension to obtain the attenuated VTT oncolytic VACA. According to the TK gene removed recombinant VTT oncolytic VACA, which is disclosed by the invention, the TK gene is used as an attenuation target, the EGFP gene and the Luciferase gene are expressed, virus virulence is reduced, and tumor selectivity of the virus is improved; the TK gene removed recombinant VTT oncolytic VACA has a function of monitoring distribution of virus particles in a body, provides a novel antitumor drug for the clinic, can be used for researching and developing a live vaccine and is applicable to various tumors such as the lung cancer, the liver cancer, the melanoma and the like.

The invention relates to a new fluoroimmunoassay method for human thymidine kinase fluoroimmunoassay based on magnetic nanometer grains. The invention sets up a quick and simple immunological detecting technology for the high flux human thymidine kinase (hTK1), and the technology is used for the hTK1 clinical examination. The hTK1 is fixed with covalence on the surface of an amido silanization superparamagnetism nanometer grain immobilized carrier through glutaric dialdehyde, a competitive immunoassay method is adopted, horse radish peroxidase is used as an enzyme labeling, ethyl-para-hydroxyphenyl acid is used as a fluorogenic substrate, thereby realizing the detecting for the hTK1. The amido silanization nucleocapsid-shaped magnetic nanometer grains are evenly dispersed in liquidoid, thereby having the advantages of fixation and uniformity, high specificity, good repeatability, quick reaction velocity, etc., under the action of an adscititious magnetic field, the invention can effectively realize the separation and enrichment of the immune complex and the reaction liquid of the nucleocapsid-shaped magnetic nanometer grains, the operation of the analytical method is simple, accurate, sensitive and fast, automatic detection is easy to be realized, and large batch of test task can be completed.

Compositions and methods useful in transgene expression are provided. Herpes simplex virus type 1 thymidine kinase sequences (“TK sequences”) are used to enhance transgene expression in first generation and high capacity adenoviral vectors. An mCMV promoter-driven β-galactosidase-expressing cassette is combined with TK sequences through direct fusion of the cDNA's. β-galactosidase (transgene) expression is enhanced independent of adenoviral vector selection. Methods of enhancing transgene expression employing the inventive adenoviral vectors are provided, along with pharmaceutical preparations comprising the inventive vectors and kits for enhanced transgene expression.

Nucleic acid sequences encoding improved Herpes Simplex Virus Thymidine Kinases are provided, including their use in diagnostic and therapeutic applications. The thymidine kinases may be mutated using conservative mutations, non-conservative mutations, or both. Also provided are gene therapeutic systems, including viral and retroviral particles.

The invention discloses a liposome medicinal composition with tumor targeting, in-vivo tracing and treating functions and a preparation method thereof. The medicinal composition is a liposome targeting medicament which is resistant to CD44 antibody coupling, has a molecular imaging function simultaneously, and can be used for monitoring the in-vivo distribution of a medicament in real time in a living body state. In particular, plasmids containing three fusion genes, including renilla luciferase, red fluorescent proteins and suicide gene thymidine kinase, are coupled to CD44 antibody mediation-resistant immunoliposome, the specificity of liposome nanoparticles in a liver cancer in-situ model of an in-vivo targeting NOD / SCID (Non-Obese Diabetic / Severe Combined Immune-Deficiency) mouse is monitored by detecting a renilla luciferase signal with a living body imaging system, and apoptosis of liver cancer cells is induced by applying target thymidine kinase of ganciclovir; and moreover, a targeted liposome can be coated with adriamycin for inducing apoptosis of the liver cancer cells. The liposome medicinal composition provided by the invention does not have any toxic or side effect, has small damage and a good effect, and is suitable to be applied for a long time.

A method of treatment of cancer, viral infections, and the like administers anti-TK1 antibody, constituted as the complete antibody or a fragment thereof. The antibody binds to the surface of cells expressing TK1 thereon. The antibody, with or without another agent bound thereto, may effect complement mediated lysis, antibody-dependent cell-mediated cell cytotoxicity, apoptosis, an immune response by the mammal, a reduction in cellular replication, a combination thereof, or the like for such cells. The antibody may be coupled to an immune response stimulator, a cytotoxin, an enzyme, a combination, or the like to effect the treatment desired.

Monoclonal antibodies to thymidine kinase 1 are disclosed which are useful in methods of detecting, diagnosing, and treating viral infection.

Methods and compositions suitable for the treatment of malignancies in subjects with a germline deletion polymorphism that blocks the activity of thymidine kinase inhibitors in triggering apoptosis in tumor cells or in subjects having a mutation in or a dysregulation of the AHI1 gene are disclosed. These methods employ an alkylating hexitol derivative such as dianhydrogalactitol, a derivative or analog of dianhydrogalactitol, diacetyldianhydrogalactitol, a derivative or analog of diacetyldianhydrogalactitol, dibromodulcitol, and a derivative or analog of dibromodulcitol. The compositions can include such alkylating hexitol derivatives. The methods can further include administration of a BH3 mimetic, and the compositions can further include a BH3 mimetic. In subjects having a dysregulation of the AHI1 gene, the methods can further include the administration of an agent modulating the expression or activity of the AHI1 gene or AHI1 protein, and the compositions can further include such an agent.

The invention discloses a recombinant oncolytic vaccinia virus, a preparation method and application thereof. A thymidine kinase TK region of a genome of a recombinant oncolytic vaccinia virus comprises an anti-mouse / human TIGIT antibody genetic sequence, and the thymidine kinase TK region of the genome of the recombinant oncolytic vaccinia virus can express an anti-mouse / human TIGIT antibody. Theanti-mouse / human TIGIT antibody genetic sequence is formed by connecting an antibody heavy chain gene, 2A peptide and an antibody light chain gene in series, and the nucleotide sequence of the anti-mouse / human TIGIT antibody gene is shown as SEQ ID NO. 1. The recombinant oncolytic vaccinia virus oncolytic virus kills tumors through direct oncolysis and activation of a human immune system. The oncolytic vaccinia virus can effectively activate the immune response of T cells to tumor cells so as to exert multiple anti-tumor effects.

Belonging to the genetic engineering field in medicine, the invention discloses a construction scheme for artificially modified human adenovirus type 5 (Ad5), and specific application of a tumor-selective replicative adenovirus construct in tumor treatment. With the techniques of PCR (polymerase chain reaction) amplifying fixed point deletion, enzyme cutting, connection, cloning, homologous recombination, transfection, single cloning purification of adenovirus and the like, a recombinant adenovirus construct can be acquired. Acquisition of the construct is characterized in that: 27 bases are deleted in 2(CR2) region of E1A conserved sequence of Ad5 genome; 28530-29360nt in 6.7k / gp19k gene in E3 region are deleted, and meanwhile a fragment of full-length cDNA(1131bp) of HSV-TK (herpes simplex virus thymidine kinase) is inserted into the deletion region. The construct of the invention is a novel oncolytic adenovirus carrier with high tumor-selective replication capability. With the suicide gene of HSV-TK and dissolubility of tumor cells by oncolytic viruses, the dual killing effects have unique utility value in tumor biotherapy.

The present invention relates to a new recombinant raccoon poxvirus vector vaccine in which the vector expresses one or more antigenic proteins encoded by multiple open reading frames, preferably the ORF5, ORF6 and / or ORF3 / ORF4 / ORF7, of one or more porcine reproductive and respiratory syndrome virus strains alone or in combination with an open reading frame of porcine circovirus type 2 (PCV-2), preferably ORF2, at the hemagglutinin (ha) and / or thymidine kinase (tk) loci.

Monoclonal antibodies to thymidine kinase 1 are disclosed which are useful in methods of detecting, diagnosing, and treating viral infection.

The present invention relates to a new recombinant raccoon poxvirus vector vaccine in which the vector expresses one or more antigenic proteins encoded by multiple open reading frames, preferably the ORF5, ORF6 and / or ORF3 / ORF4 / ORF7, of one or more porcine reproductive and respiratory syndrome virus strains alone or in combination with an open reading frame of porcine circovirus type 2 (PCV-2), preferably ORF2, at the hemagglutinin (ha) and / or thymidine kinase (tk) loci.