EP0334301A1
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Recombinant retroviruses
Assignee: Novartis Vaccines and Diagnostics Inc
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Filing: 1989-03-21
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Granted: —
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Expires: 2009-03-21
Abstract
Abstract Recombinant retroviruses carrying a vector construct capable of preventing, inhibiting, stabilizing or reversing infectious, cancerous or auto-immune diseases are disclosed. More specifically, the recombinant retroviruses of the present invention are useful for (a) stimulating a specific immune response to an antigen or a pathogenic antigen; (b) inhibiting a function of a pathogenic agent, such as a virus; and (c) inhibiting the interaction of an agent with a host cell receptor. In addition, eucaryotic cells infected with, and pharmaceutical compositions containing such a recombinant retrovirus are disclosed. Various methods for producing recombinant retroviruses having unique characteristics, and methods for producing transgenic packaging animals or insects are also disclosed.
Inventors
- 👤 Harry E. Gruber
- 👤 Douglas J. Jolly
- 👤 James G. Respess
- 👤 Paul K. Laikind
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-08 03:06
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/79; C12N15/85; C12N15/86; A; A61; A61K; A61K38/00; A61K38/16; A61K38/17; A61K38/19; A61K38/20; A61K38/2013; A61K38/21; A61K38/217; A61P; A61P25/00; A61P25/28; A61P3/00; A61P3/08; A61P3/10; A61P31/00; A61P31/12; A61P31/14; A61P31/18; A61P35/00; A61P37/00; A61P37/02; A61P9/00; C07; C07K; C07K14/00; C07K14/005; C07K14/435; C07K14/705; C07K14/70503; C07K14/70514; C12N7/00; C12Q; C12Q1/00; C12Q1/02; A01; A01K; A01K2217/00; A01K2217/05; A61K39/00; A61K2039/51; A61K2039/525; A61K2039/5256; A61K2039/53; A61K2039/57; A61K48/00; C12N2740/00; C12N2740/00011; C12N2740/10011; C12N2740/10041; C12N2740/10043; C12N2740/13011; C12N2740/13022; C12N2740/16011; C12N2740/16022; C12N2740/16023; C12N2740/16041; C12N2740/16043; C12N2740/16111; C12N2740/16122; C12N2740/16211; C12N2740/16222
Claims Analysis
Total Claims: 66
Claim 1
Independent Claim
19 words
A recombinant retrovirus carrying a vector construct capable of preventing, inhibiting, stabilizing or reversing infectious, cancerous or auto-immune diseases.
Claim 2
Dependent Claim
19 words
A recombinant retrovirus carrying a vector construct capable of preventing, inhibiting, stabilizing or reversing infectious, cancerous or auto-immune diseases.
Claim 3
Dependent Claim
26 words
The recombinant retrovirus of claim 1 wherein the vector construct directs the expression of a protein or portion thereof in target cells infected with said retrovirus.
Claim 4
Dependent Claim
43 words
The recombinant retrovirus of claim 1 wherein the vector construct directs the expression of an antigen or modified form thereof in target cells infected with the retrovirus, said antigen or modified form thereof being capable of stimulating an immune response within an animal.
Claim 5
Dependent Claim
50 words
The recombinant retrovirus of claim 1 wherein the vector construct directs the expression of a modified form of a pathogenic antigen in target cells infected with the retrovirus, said modified antigen being capable of stimulating an immune response within an animal but having reduced pathogenicity relative to the pathogenic antigen.
Claim 6
Dependent Claim
17 words
The recombinant retrovirus of claims 3 or 4 wherein the expressed antigen elicits a cell-mediated immune response.
Claim 7
Dependent Claim
23 words
The recombinant retrovirus of claims 3 or 4 wherein the expressed antigen elicits an HLA class I- and/or class II- restricted immune response.
Claim 8
Dependent Claim
31 words
The recombinant retrovirus of claims 3-6, wherein the expressed antigen is an HIV protein selected from the group consisting of gp 160, gp 120 and gp 41, or modified forms thereof.
Claim 9
Dependent Claim
26 words
The recombinant retrovirus of claim 1 wherein the vector construct directs the expression of a specific T-cell receptor or immunoglobulin that recognizes an antigen of interest.
Claim 10
Dependent Claim
37 words
The recombinant retrovirus of claim 1 wherein the vector construct directs the expression of a palliative in cells infected with the retrovirus, said palliative being capable of inhibiting a function of a pathogenic agent necessary for pathogenicity.
Claim 11
Dependent Claim
37 words
The recombinant retrovirus of claim 9 wherein the pathogenic agent is a virus, and the inhibited function is selected from the group consisting of absorption, replication, gene expression, assembly, and exit of the virus from infected cells.
Claim 12
Dependent Claim
41 words
The recombinant retrovirus of claim 9 wherein the pathogenic agent is a cancerous cell or cancer-promoting growth factor, and the inhibited function is selected from the group consisting of cell replication, susceptibility to external signals, and lack of production of anti-oncogenes.
Claim 13
Dependent Claim
37 words
The recombinant retrovirus of claim 9 wherein the vector construct directs the expression of a toxic palliative in infected target cells in response to the presence in said cells of an entity associated with the pathogenic agent.
Claim 14
Dependent Claim
23 words
The recombinant retrovirus of claim 9 wherein the palliative is capable of selectively inhibiting the expression or the effects of a pathogenic gene.
Claim 15
Dependent Claim
17 words
The recombinant retrovirus of claim 13 wherein the palliative comprises an inhibiting peptide specific for viral protease.
Claim 16
Dependent Claim
19 words
The recombinant retrovirus of claim 9 wherein the palliative comprises antisense RNA complementary to RNA sequences necessary for pathogenicity.
Claim 17
Dependent Claim
23 words
The recombinant retrovirus of claim 9 wherein the palliative promotes recombination or integration into a pathogenic genome, so as to disrupt its function.
Claim 18
Dependent Claim
29 words
The recombinant retrovirus of claim 9 wherein the palliative comprises a defective structural protein of a pathogenic agent, said protein being capable of inhibiting assembly of the pathogenic agent.
Claim 19
Dependent Claim
18 words
The recombinant retrovirus of claim 9 wherein the vector construct directs the expression of the HSVTK gene product.
Claim 20
Dependent Claim
18 words
The recombinant retrovirus of claim 9 wherein the vector construct directs the expression of a tumor supressor gene.
Claim 21
Dependent Claim
44 words
The recombinant retrovirus of claim 9 wherein the vector construct directs the expression of a protein that activates a compound with little or no cytotoxicity into a toxic product in the presence of a pathogenic agent, thereby effecting localized therapy to the pathogenic agent.
Claim 22
Dependent Claim
13 words
The recombinant retrovirus of claim 20 wherein said protein is herpes thymidine kinase.
Claim 23
Dependent Claim
11 words
The recombinant retrovirus of claim 20 wherein said protein is CD4.
Claim 24
Dependent Claim
31 words
The recombinant retrovirus of claim 9 wherein the vector construct directs the expression of a protein that is toxic upon processing or modification by a protein derived from a pathogenic agent.
Claim 25
Dependent Claim
29 words
The recombinant retrovirus of claim 9 wherein the vector construct expresses a reporting product on the surface of target cells infected with the retrovirus and containing the pathogenic agent.
Claim 26
Dependent Claim
29 words
The recombinant retrovirus of claim 1 wherein the vector construct directs the expression of a gene capable of suppressing the immune system in target cells infected with said retrovirus.
Claim 27
Dependent Claim
44 words
The recombinant retrovirus of claim 1 wherein the vector construct directs the expression of a blocking element in cells infected with said retrovirus, said blocking element being capable of binding to either a receptor or an agent such that the receptor/agent interaction is blocked.
Claim 28
Dependent Claim
45 words
The recombinant retrovirus of claim 1 wherein the vector construct directs the expression of a blocking element in cells infected with said retrovirus, said blocking element being capable of binding to a receptor or an envelope protein, such that the receptor/envelope protein interaction is blocked.
Claim 29
Dependent Claim
18 words
The recombinant retrovirus of claims 26 or 27 wherein the blocking element is secreted from the infected cells.
Claim 30
Dependent Claim
14 words
The recombinant retrovirus of any of claims 1-28, wherein said retrovirus is replication defective.
Claim 31
Dependent Claim
30 words
A method of producing a recombinant retrovirus, comprising: packaging a vector construct in a capsid and envelope such that a replication defective recombinant retrovirus according to claim 29 is produced.
Claim 32
Dependent Claim
14 words
Ex vivo cells infected with a recombinant retrovirus according to any of claims 1-29.
Claim 33
Dependent Claim
28 words
Eucaryotic cells infected with a recombinant retrovirus according to any of claims 1-28, said cells being capable of generating infectious particles containing any one of said vector constructs.
Claim 34
Dependent Claim
22 words
A pharmaceutical composition comprising a retrovirus according to any one of claims 1-29, in combination with a physiologically acceptable carrier or diluent.
Claim 35
Dependent Claim
13 words
The pharmaceutical composition of claim 33, for use as an active therapeutic substance.
Claim 36
Independent Claim
84 words
A method of testing a sample for the presence of a virus carrying a particular gene, said gene being capable of expressing an identifying protein in indicator cells having (1) a marker gene which is capable of expressing a marker product; and (2) control sequences which respond to the presence of the identifying protein by switching expression of the marker gene between expressing and non-expressing states; the method comprising contacting indicator cells with the sample, and testing for the presence of the marker product.
Claim 37
Independent Claim
84 words
A method of testing a sample for the presence of a virus carrying a particular gene, said gene being capable of expressing an identifying protein in indicator cells having (1) a marker gene which is capable of expressing a marker product; and (2) control sequences which respond to the presence of the identifying protein by switching expression of the marker gene between expressing and non-expressing states; the method comprising contacting indicator cells with the sample, and testing for the presence of the marker product.
Claim 38
Independent Claim
96 words
A method of testing for the presence of a particular gene in a cell sample, said gene being capable of expressing an identifying protein, comprising: infecting cells within the sample with a recombinant retrovirus comprising a vector construct coding for (1) a marker gene which is capable of expressing a marker product in the cell sample; and (2) control sequences which respond to the presence of the identifying protein in the cells within the sample by switching expression of the marker gene between expressing and non-expressing states; and testing for the presence of the marker product.
Claim 39
Independent Claim
96 words
A method of testing for the presence of a particular gene in a cell sample, said gene being capable of expressing an identifying protein, comprising: infecting cells within the sample with a recombinant retrovirus comprising a vector construct coding for (1) a marker gene which is capable of expressing a marker product in the cell sample; and (2) control sequences which respond to the presence of the identifying protein in the cells within the sample by switching expression of the marker gene between expressing and non-expressing states; and testing for the presence of the marker product.
Claim 40
Independent Claim
27 words
The method of claims 35 or 36 wherein the control sequences switch expression of the marker product by switching the marker gene between transcribing and nontranscribing states.
Claim 41
Dependent Claim
79 words
An indicator cell suitable for testing for the presence of a virus carrying a particular gene which is capable of expressing an identifying protein in the indicator cell, the indicator cell having a genome coding for (1) a marker gene which can express a marker product in the indicator cell; and (2) control sequences which respond to the presence of the identifying protein in the indicator cell by switching expression of the marker gene between expressing and non-expressing states.
Claim 42
Dependent Claim
79 words
An indicator cell suitable for testing for the presence of a virus carrying a particular gene which is capable of expressing an identifying protein in the indicator cell, the indicator cell having a genome coding for (1) a marker gene which can express a marker product in the indicator cell; and (2) control sequences which respond to the presence of the identifying protein in the indicator cell by switching expression of the marker gene between expressing and non-expressing states.
Claim 43
Dependent Claim
71 words
A recombinant retrovirus suitable for infecting cells and testing for the presence of a particular gene therein which produces an identifying protein, the recombinant retrovirus having a genome coding for: (1) a marker gene which can express a marker product in the cells; and (2) control sequences which respond to the presence of the identifying protein in the cells by switching expression of the marker gene between expressing and non-expressing states.
Claim 44
Dependent Claim
71 words
A recombinant retrovirus suitable for infecting cells and testing for the presence of a particular gene therein which produces an identifying protein, the recombinant retrovirus having a genome coding for: (1) a marker gene which can express a marker product in the cells; and (2) control sequences which respond to the presence of the identifying protein in the cells by switching expression of the marker gene between expressing and non-expressing states.
Claim 45
Independent Claim
110 words
A method of selecting packaging cells which produce high levels of a primary agent selected from a packaging protein and a gene product of interest, comprising: (a) providing in packaging cells a genome comprising a primary gene which expresses a primary agent therein, and a selectable gene which expresses a selectable protein therein at lower levels than the primary agent, the expression levels of the primary gene and selectable gene being proportional; (b) exposing the packaging cells to a selecting agent which enables identification of those cells which express the selectable protein at a critical level; and (c) detecting those packaging cells which express high levels of the primary agent.
Claim 46
Independent Claim
110 words
A method of selecting packaging cells which produce high levels of a primary agent selected from a packaging protein and a gene product of interest, comprising: (a) providing in packaging cells a genome comprising a primary gene which expresses a primary agent therein, and a selectable gene which expresses a selectable protein therein at lower levels than the primary agent, the expression levels of the primary gene and selectable gene being proportional; (b) exposing the packaging cells to a selecting agent which enables identification of those cells which express the selectable protein at a critical level; and (c) detecting those packaging cells which express high levels of the primary agent.
Claim 47
Independent Claim
62 words
A method of producing a recombinant retrovirus, comprising: generating gag, pol, and env proteins from a cell line infected by a recombinant virus which is capable of producing the proteins; and contacting the proteins with viral vector RNA, tRNA, liposomes, and a cell extract to complement missing functions for particle assembly, so as to produce recombinant retroviruses carrying the viral vector RNA.
Claim 48
Independent Claim
62 words
A method of producing a recombinant retrovirus, comprising: generating gag, pol, and env proteins from a cell line infected by a recombinant virus which is capable of producing the proteins; and contacting the proteins with viral vector RNA, tRNA, liposomes, and a cell extract to complement missing functions for particle assembly, so as to produce recombinant retroviruses carrying the viral vector RNA.
Claim 49
Independent Claim
47 words
A method of producing a recombinant retrovirus, comprising: (a) generating recombinant viral vectors which separately or in combination, code for gag/pol, env and a retroviral vector genome; (b) producing high titre stocks of the vectors; and (c) co-infecting primary or other cells to generate recombinant retroviral vectors.
Claim 50
Independent Claim
47 words
A method of producing a recombinant retrovirus, comprising: (a) generating recombinant viral vectors which separately or in combination, code for gag/pol, env and a retroviral vector genome; (b) producing high titre stocks of the vectors; and (c) co-infecting primary or other cells to generate recombinant retroviral vectors.
Claim 51
Independent Claim
79 words
A method of producing a retrovirus, comprising growing a producer cell having a genome comprising: (a) a gene of interest along with a packaging signal of a first retroviral phenotype; (b) gag and pol genes of the first retroviral phenotype, absent a packaging signal; (c) a hybrid env gene absent a packaging signal, the product of said hybrid env gene comprising a cytoplasmic segment of the first retroviral phenotype, and a binding segment exogenous to the first retroviral phenotype.
Claim 52
Independent Claim
79 words
A method of producing a retrovirus, comprising growing a producer cell having a genome comprising: (a) a gene of interest along with a packaging signal of a first retroviral phenotype; (b) gag and pol genes of the first retroviral phenotype, absent a packaging signal; (c) a hybrid env gene absent a packaging signal, the product of said hybrid env gene comprising a cytoplasmic segment of the first retroviral phenotype, and a binding segment exogenous to the first retroviral phenotype.
Claim 53
Dependent Claim
58 words
A hybrid env gene useful for preparing a retrovirus which can selectively carry a gene of interest to a target cell, the env gene coding for: (a) a cytoplasmic segment of a first retroviral phenotype; and (b) a binding segment exogenous to the first retroviral phenotype, the binding segment being capable of selectively binding to the target cell.
Claim 54
Dependent Claim
58 words
A hybrid env gene useful for preparing a retrovirus which can selectively carry a gene of interest to a target cell, the env gene coding for: (a) a cytoplasmic segment of a first retroviral phenotype; and (b) a binding segment exogenous to the first retroviral phenotype, the binding segment being capable of selectively binding to the target cell.
Claim 55
Dependent Claim
70 words
A recombinant retrovirus carrying a vector construct, comprising: (a) an exogenous gene to be inserted into a preselected site on a target cell's genome; and (b) a segment substantially homologous to a corresponding sequence of the target cell's genome adjacent to the preselected site, said sequence being of a sufficient length such that homologous recombination takes place between the vector construct and the target cell's genome at the preselected site.
Claim 56
Dependent Claim
70 words
A recombinant retrovirus carrying a vector construct, comprising: (a) an exogenous gene to be inserted into a preselected site on a target cell's genome; and (b) a segment substantially homologous to a corresponding sequence of the target cell's genome adjacent to the preselected site, said sequence being of a sufficient length such that homologous recombination takes place between the vector construct and the target cell's genome at the preselected site.
Claim 57
Independent Claim
76 words
A method of inserting an exogenous gene into a preselected site on a target cell's genome, the method comprising exposing the target cells to a recombinant retrovirus having a genome comprising: (a) an exogenous gene; (b) a segment substantially homologous to a corresponding sequence of the target cell's genome adjacent the preselected site, which sequence is sufficiently long as to facilitate homologous recombination between the vector construct and the target cell's genome at the preselected site.
Claim 58
Independent Claim
76 words
A method of inserting an exogenous gene into a preselected site on a target cell's genome, the method comprising exposing the target cells to a recombinant retrovirus having a genome comprising: (a) an exogenous gene; (b) a segment substantially homologous to a corresponding sequence of the target cell's genome adjacent the preselected site, which sequence is sufficiently long as to facilitate homologous recombination between the vector construct and the target cell's genome at the preselected site.
Claim 59
Independent Claim
55 words
A method of producing a recombinant retrovirus which is capable of integrating its genome into a preselected site on a target cell's genome, comprising: packaging a vector in a capsid and envelope, and including in the viral particle a modified form of integrase which is capable of integrating the retroviral genome into the preselected site.
Claim 60
Independent Claim
55 words
A method of producing a recombinant retrovirus which is capable of integrating its genome into a preselected site on a target cell's genome, comprising: packaging a vector in a capsid and envelope, and including in the viral particle a modified form of integrase which is capable of integrating the retroviral genome into the preselected site.
Claim 61
Independent Claim
60 words
A method of producing recombinant retroviruses, comprising: mating a transgenic animal or insect containing a gag/pol-env viral construct, with a transgenic animal or insect containing a vector construct containing a promoter; isolating the progeny of said transgenic animals or insects; isolating selected cells from the progeny; growing said cells in an appropriate medium; and isolating recombinant retroviruses from the cells.
Claim 62
Independent Claim
60 words
A method of producing recombinant retroviruses, comprising: mating a transgenic animal or insect containing a gag/pol-env viral construct, with a transgenic animal or insect containing a vector construct containing a promoter; isolating the progeny of said transgenic animals or insects; isolating selected cells from the progeny; growing said cells in an appropriate medium; and isolating recombinant retroviruses from the cells.
Claim 63
Independent Claim
61 words
A method of producing a transgenic packaging animal or insect, comprising: mating a transgenic animal or insect containing a vector construct coding for some, but not all viral proteins necessary for packaging, with a transgenic animal or insect containing a vector construct coding for the remainder of said necessary viral proteins; and isolating the progeny of said transgenic animals or insects.
Claim 64
Independent Claim
61 words
A method of producing a transgenic packaging animal or insect, comprising: mating a transgenic animal or insect containing a vector construct coding for some, but not all viral proteins necessary for packaging, with a transgenic animal or insect containing a vector construct coding for the remainder of said necessary viral proteins; and isolating the progeny of said transgenic animals or insects.
Claim 65
Dependent Claim
34 words
The method of claim 49, further comprising the step of mating said progeny with a transgenic animal or insect containing a vector construct, to produce primary cells capable of producing high titre recombinant retrovirus.
Claim 66
Dependent Claim
35 words
The method of claim 49, further comprising the step of infecting cells explanted from said progeny with a recombinant retrovirus containing a vector construct to produce primary cells capable of producing high titre recombinant retrovirus.
References Cited (Backward Citations)
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