WO1997029201A2 - Vecteur retroviral pour le transfert genique d'un antagoniste d'il6 dans des cellules souches hematopoietiques humaines - Google Patents

Vecteur retroviral pour le transfert genique d'un antagoniste d'il6 dans des cellules souches hematopoietiques humaines Download PDF

Info

Publication number
WO1997029201A2
WO1997029201A2 PCT/DE1997/000231 DE9700231W WO9729201A2 WO 1997029201 A2 WO1997029201 A2 WO 1997029201A2 DE 9700231 W DE9700231 W DE 9700231W WO 9729201 A2 WO9729201 A2 WO 9729201A2
Authority
WO
WIPO (PCT)
Prior art keywords
gene
vector
antagonist
retroviral
vector according
Prior art date
Application number
PCT/DE1997/000231
Other languages
German (de)
English (en)
Other versions
WO1997029201A3 (fr
Inventor
Ralf Bargou
Christopher Baum
Dirk HÖNEMANN
Original Assignee
Max-Delbrück-Centrum für Molekulare Medizin
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Max-Delbrück-Centrum für Molekulare Medizin filed Critical Max-Delbrück-Centrum für Molekulare Medizin
Publication of WO1997029201A2 publication Critical patent/WO1997029201A2/fr
Publication of WO1997029201A3 publication Critical patent/WO1997029201A3/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • C12N15/86—Viral vectors
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/52—Cytokines; Lymphokines; Interferons
    • C07K14/54—Interleukins [IL]
    • C07K14/5412—IL-6
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2740/00—Reverse transcribing RNA viruses
    • C12N2740/00011—Details
    • C12N2740/10011—Retroviridae
    • C12N2740/13011—Gammaretrovirus, e.g. murine leukeamia virus
    • C12N2740/13041—Use of virus, viral particle or viral elements as a vector
    • C12N2740/13043—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2840/00—Vectors comprising a special translation-regulating system
    • C12N2840/20—Vectors comprising a special translation-regulating system translation of more than one cistron
    • C12N2840/203—Vectors comprising a special translation-regulating system translation of more than one cistron having an IRES

Definitions

  • the invention relates to a retroviral vector for the gene transfer of an IL6 antagonist into human hematopoietic stem cells for the treatment of multiple myeloma.
  • Areas of application of the invention are medicine and the pharmaceutical industry.
  • Plasmacytoma Multiple myeloma, or plasmacytoma, is an incurable malignant disease of the antibody-secreting B cell with an annual incidence of 4 new diseases per 100,000 inhabitants in Germany.
  • the main localization of the plasma cytoma cells is bone and bone marrow, where they lead to osteolysis or displacement of the hematopoiesis, with the consequences anemia, leukocytopenia and thrombocytopenia.
  • the median survival after the onset of the disease is 3 years (W. Siegenthaler et al., Textbook of Internal Medicine, 3rd edition (1992), pp. 723-727).
  • interleukin-6 has been characterized as a cytokine that is of crucial importance for the growth of plasma cytoma cells. It has recently been shown that it is not possible to induce plasmacytomas or to propagate already established (syngeneic) plasmacytoma cells in transgenic IL6 "knock-out" mice, that is to say mice that no longer have a functional IL6 gene not - ' ⁇ nock-out "mice was readily possible.
  • IL6 interleukin-6
  • the aim of the invention is to develop a gene therapy strategy for combating multiple myeloma with the aid of IL6 antagonists.
  • the aim of this strategy is to ensure that the biologically active concentration of this antagonist in the bone marrow is sufficiently high in clinical use to neutralize the IL6 present.
  • the invention is therefore based on the object of constructing a vector which, after transfer to autologous hematopoietic stem cells, brings about a strong and stable expression of a suitable IL6 antagonist.
  • the essential basis of the invention is a retroviral starting vector, preferably a MoMLV derivative, particularly preferably the vector pSFlN, which shows a particularly high level of expression for gene transfer into hematopoietic stem and progenitor cells.
  • the gene of an IL6 antagonist and a suicide or a selection gene are incorporated into this vector using methods known per se in genetic engineering.
  • the thymidine kinase gene and the cytosine desaminase gene are the main suicide genes suitable.
  • the puromycin gene, the neomycin acetyltransferase gene or the mdr-1 gene are preferably used as the selection gene.
  • the gene of the IL6 antagonist and the suicide or selection gene are preferably located on the same mRNA, that is to say they are controlled in expression by the same promoter which is particularly suitable for stem cells, and are nevertheless translated separately.
  • the suicide gene unforeseen side effects of the therapeutic gene, e.g. on the normal hematopoiesis of the patient, those cells are produced which produce the IL6 antagonist.
  • stem cells which produce high amounts of mdr-1 under the selection pressure of chemotherapy will also express large amounts of IL6 antagonist at the same time.
  • the selection of the IL6 antagonist is of particular importance for the effect of the vector.
  • all known IL6 antagonists can be used, but preference is given to up to 20 codons over the human interleukin-6 mutant antagonists.
  • Another important embodiment is an IL6 antagonist containing 12 codon mutations
  • ACATGTGTGA 251 AAGCAGCAAA GAGGCACTGG CAGAAAACAA CCTGAACCTT
  • CTCATTCTGC 601 GCAGCTTTAA GGAGTTCCTG atccgCAGCC TGAGGGCTCT
  • SV40 Simian Virus 40 early promoter
  • the vector is packaged using known viral vector packaging lines, which can generate non-replication-capable retroviruses.
  • the selection of the virus envelope used has a significant impact on the efficiency of the gene transfer.
  • the lines FLY13 and GP + envAml2 are particularly suitable.
  • the vector is used in such a way that hematopoietic stem cells from peripheral blood or bone marrow of patients are stimulated by cytokine, the vector is transferred into these stem cells and they are then returned to the patient, preferably by intravenous injection. Since hematopoietic stem cells preferentially ' homen ' in the bone marrow after systemic administration, the expression of the IL6 antagonist is ensured in this organ.
  • the vector according to the invention has a high gene expression rate in hematopoietic stem cells and can therefore be used effectively. It enables a curative approach to the treatment of multiple myeloma, which is potentially superior to previous therapies.
  • the invention will be explained in more detail below by application examples.
  • This vector is a MoLMV derivative and contains regulatory elements of the viruses FUCF and MESV, which lead to the fact that the genes transferred with this vector pSFlN are particularly strongly expressed in hematopoietic stem and progenitor cells.
  • This vector is therefore superior to previous vectors with regard to gene transfer in hematopoietic stem cells and is therefore particularly suitable for a possible gene therapy use of the IL6 antagonist in hematopoietic Staram cells.
  • the antagonist was equipped with a myc tag. In this way it is easily possible to detect the IL6 antagonist expression in the serum or bone marrow of the patient by means of immunological methods (eg ELISA).
  • the neomycin resistance gene in the pSFlN was replaced by a puromycin resistance gene and linked to the IL6 antagonist via an IRES sequence.
  • the new vector resulting from the incorporation of the IL6 antagonist is called pSF-IL-6AP.
  • a suicide gene the herpes simplex thymidine kinase, was incorporated into the vector in a further step.
  • This enables the cell genetically modified by the vector to be killed in the patient by systemic administration of ganciclovir.
  • both cDNAs were linked in the vector via an IRES (inter-ribosomal recognition site) sequence.
  • IRES internal-ribosomal recognition site
  • both genes are located on the same mRNA, that is to say they are controlled in expression by the same promoter which is particularly suitable for stem cells, and are nevertheless translated separately.
  • the new vector resulting from the incorporation of the Tk suicide gene is called pSF-IL-6AT.
  • the multidrug resistance gene (radr-1) was additionally incorporated into the vector.
  • chemotherapy which is used in the context of conventional plasmacytoma therapy, can enrich those stem cells in the patient that also carry the IL6 antagonist. This would increase the IL6 antagonist concentration in the patient and increase the therapeutic efficacy of the vector.
  • both cDNAs in the vector were linked via an IRES (inter-ribosomal recognition site) sequence. In this way, both genes are located on the same raRNA, that is to say they are controlled in expression by the same promoter which is particularly suitable for stem cells, and are nevertheless translated separately. Stem cells that express high amounts of mdr-1 under the pressure of chemotherapy will also simultaneously express large amounts of IL6 antagonist.
  • the new vector resulting from the incorporation of the mdr-1 gene is called pSF-IL-6AM.
  • the vector DNA is packaged into viruses using the retroviral packaging cell lines FLY13 and FLYRD18.
  • These packaging lines have the advantage that they generate much higher (100-fold) virus titers than previously used lines (Cosset et al., Journ. Virol. 69: 7430-36, 1995). This leads to an improvement in gene transfer efficiency in hematopoietic stem cells.
  • Another advantage of these lines is that they produce viral vectors that are stable in human serum. Since gene transfer must be carried out in human serum as a cultivation medium in a clinical application, these packaging lines are very well suited for this purpose.
  • the virus-containing supernatant from the packaging lines is used in gene transfer to primary hematopoietic stem cells.
  • the stem cells are previously treated with a cytokine cocktail (Brugger et al., Blood 81: 2579-84, 1993) for 24 hours. The virus supernatant is then added and cultivated for 48 hours. This leads to a gene transfer efficiency of 10-20%. 6)
  • the application of the invention is primarily intended for the treatment of multiple myeloma. Since it has been shown that the cytokine IL6 is an essential growth factor for plasmotytom cells and the withdrawal of this factor leads to the death of the malignant cells, the expression of high concentrations of the IL6 antagonist in the bone marrow by means of gene transfer in stem cells represents a curative approach to the treatment of multiple myeloma and is therefore potentially superior to previous therapy methods.
  • Item 57 (aspartate for leucine), item 59 (phenylalanine for glutamate), item 60 (tryptophan for asparagine).
  • Pos. 75 (tyrosine for glutamine), Pos. 76 (lysine for serine)
  • Steps 2-4 take place analogously to example I.
  • the vector DNA is packaged into viruses using the retroviral packaging line GP + envAml2 (Markowitz et al., Virology 167, 400-406, 1988).
  • This packaging line has the advantage that the titers are not significantly lower than those of the fly
  • the virus envelope used here is particularly suitable for the infection of hematopoietic stem cells (Xu et al., Exp.- Hematology 22 (2), Feb 223-30, 1994).
  • this vector also contains another internal promoter (SV 40 or CMV) for the independent transcription of the resistance gene, in this case the gene for neomycin resistance.
  • This proven vector design allows the in vitro selection of infected stem cells using neomycin or neomycin analogs and has already been used successfully in several stem cell gene therapy studies (e.g.
  • retroviral vectors are packaged analogously to Examples I and II in the packaging cell lines used there, and the infection protocol does not change (with the exception of the use of neomycin as a selection antibiotic).

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biophysics (AREA)
  • Wood Science & Technology (AREA)
  • Molecular Biology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Physics & Mathematics (AREA)
  • Toxicology (AREA)
  • Virology (AREA)
  • Plant Pathology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Microbiology (AREA)
  • Medicinal Chemistry (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

L'invention concerne un vecteur rétroviral pour le transfert génique d'un antagoniste d'interleukine 6(IL6) dans des cellules souches hématopoïétiques humaines en vue du traitement du myélome multiple. Les domaines d'application de l'invention sont la médecine et l'industrie pharmaceutique. Le vecteur décrit est constitué d'un vecteur initial rétroviral, du gène d'un antagoniste d'IL6, d'un gène-suicide ou d'un gène de sélection, et éventuellement de promoteurs pour la transcription du gène-suicide ou du gène de sélection. Une forme préférée de réalisation est constituée du dérivé pLXSN de MoMLV comme vecteur initial rétroviral, du gène d'un antagoniste d'IL6 ayant muté, par rapport à l'interleukine 6 humaine, au niveau de 20 codons maximum, du gène pour la néomycine acétyl-transférase comme gène de sélection, et des promoteurs précoces immédiats viraux des virus CMV et SV40. Une autre forme de réalisation préférée est constituée du dérivé pSF1N de MoMLV comme vecteur initial rétroviral, du gène d'un antagoniste d'IL6 ayant muté, par rapport à l'interleukine 6 humaine, au niveau de 7 codons, et du gène de thymidine-kinase ou bien du gène de cytosine-désaminase comme gène-suicide ou encore du gène de puromycine ou du gène de mdr-1 comme gène de sélection.
PCT/DE1997/000231 1996-02-07 1997-01-29 Vecteur retroviral pour le transfert genique d'un antagoniste d'il6 dans des cellules souches hematopoietiques humaines WO1997029201A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE19604378 1996-02-07
DE19604378.6 1996-02-07

Publications (2)

Publication Number Publication Date
WO1997029201A2 true WO1997029201A2 (fr) 1997-08-14
WO1997029201A3 WO1997029201A3 (fr) 1997-10-02

Family

ID=7784718

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/DE1997/000231 WO1997029201A2 (fr) 1996-02-07 1997-01-29 Vecteur retroviral pour le transfert genique d'un antagoniste d'il6 dans des cellules souches hematopoietiques humaines

Country Status (2)

Country Link
DE (1) DE19704979A1 (fr)
WO (1) WO1997029201A2 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002020060A1 (fr) * 2000-09-08 2002-03-14 Phogen Limited Agregats composes de la proteine vp22 et d'acides nucleiques et utilisation de tels agregats
US6551587B2 (en) 1994-11-28 2003-04-22 Genetic Therapy, Inc. Vectors for tissue-specific replication
US6638762B1 (en) 1994-11-28 2003-10-28 Genetic Therapy, Inc. Tissue-vectors specific replication and gene expression

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE60003632T2 (de) * 1999-01-11 2004-04-15 Leadd B.V. Verwendung von apoptose-induzierende mittel zur herstellung eines mittels zur behandlung von (auto)immunkrankheiten

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993001281A1 (fr) * 1991-07-03 1993-01-21 The United States Of America, Represented By The Secretary, Department Of Health And Human Services Systeme de selection negative de cytosine desaminase pour techniques et therapies de transfert de gene
WO1994024870A1 (fr) * 1993-01-20 1994-11-10 Biotransplant, Inc. Vecteurs retroviraux capables d'exprimer des proteines multimeres a partir de sites multiples d'initiation de traduction
WO1995032282A1 (fr) * 1994-05-19 1995-11-30 Human Genome Sciences, Inc. Variant d'epissure d'interleukine-6
WO1995034669A2 (fr) * 1994-06-02 1995-12-21 Somatix Therapy Corporation Vecteurs retroviraux de therapie genique et procedes therapeutiques correspondants
WO1996007747A1 (fr) * 1994-09-08 1996-03-14 Boehringer Mannheim Gmbh Hybrides vectoriels retroviraux et leur utilisation pour le transfert de genes
WO1996034104A1 (fr) * 1995-04-28 1996-10-31 Istituto Di Ricerche Di Biologia Molecolare P. Angeletti S.P.A. Antagonistes de l'interleukine-6 humaine totalement incapables de se lier a la gp130 et leur utilisation pour l'elaboration de composes pharmaceutiques

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993001281A1 (fr) * 1991-07-03 1993-01-21 The United States Of America, Represented By The Secretary, Department Of Health And Human Services Systeme de selection negative de cytosine desaminase pour techniques et therapies de transfert de gene
WO1994024870A1 (fr) * 1993-01-20 1994-11-10 Biotransplant, Inc. Vecteurs retroviraux capables d'exprimer des proteines multimeres a partir de sites multiples d'initiation de traduction
WO1995032282A1 (fr) * 1994-05-19 1995-11-30 Human Genome Sciences, Inc. Variant d'epissure d'interleukine-6
WO1995034669A2 (fr) * 1994-06-02 1995-12-21 Somatix Therapy Corporation Vecteurs retroviraux de therapie genique et procedes therapeutiques correspondants
WO1996007747A1 (fr) * 1994-09-08 1996-03-14 Boehringer Mannheim Gmbh Hybrides vectoriels retroviraux et leur utilisation pour le transfert de genes
WO1996034104A1 (fr) * 1995-04-28 1996-10-31 Istituto Di Ricerche Di Biologia Molecolare P. Angeletti S.P.A. Antagonistes de l'interleukine-6 humaine totalement incapables de se lier a la gp130 et leur utilisation pour l'elaboration de composes pharmaceutiques

Non-Patent Citations (8)

* Cited by examiner, † Cited by third party
Title
BLOOD, Bd. 84, Nr. 9, 1.November 1994, Seiten 2890-2897, XP000196799 KALLE VON C ET AL: "INCREASED GENE TRANSFER INTO HUMAN HEMATOPOIETIC PROGENITOR CELLS BY EXTENDED IN VITRO EXPOSURE TO A PSEUDOTYPED RETROVIRAL VECTOR" *
BLOOD, Bd. 87, Nr. 11, 1.Juni 1996, Seiten 4510-4519, XP000612195 SPORENO E ET AL: "HUMAN INTERLEUKIN-6 RECEPTOR SUPER-ANTAGONISTS WITH HIGH POTENCY AND WIDE SPECTRUM ON MULTIPLE MYELOMA CELLS" *
EMBO JOURNAL, Bd. 13, Nr. 24, 1994, Seiten 5863-5870, XP000606356 SAVINO R ET AL: "RATIONAL DESIGN OF A RECEPTOR SUPER-ANTAGONIST OF HUMAN INTERLEUKIN-6" in der Anmeldung erw{hnt *
GENE, Bd. 137, Nr. 1, 1993, Seiten 139-143, XP002036890 CRAVCHIK A AND MATUS A: "A NOVEL STRATEGY FOR THE IMMUNOLOGICAL TAGGING OF cDNA CONSTRUCTS" *
JOURNAL OF EXPERIMENTAL MEDICINE, Bd. 180, Nr. 6, 1.Dezember 1994, Seiten 2395-2400, XP000601681 HON DE F D ET AL: "DEVELOPMENT OF AN INTERLEUKIN (IL) 6 RECEPTOR ANTAGONIST THAT INHIBITS IL-6-DEPENDENT GROWTH OF HUMAN MYELOMA CELLS" *
JOURNAL OF VIROLOGY, Bd. 69, Nr. 12, Dezember 1995, Seiten 7541-7547, XP002036889 BAUM C ET AL: "NOVEL RETROVIRAL VECTORS FOR EFFICIENT EXPRESSION OF THE MULTIDRUG RESISTANCE (mdr-1) GENE IN EARLY HEMATOPOIETIC CELLS" in der Anmeldung erw{hnt *
NUCLEIC ACIDS RESEARCH, Bd. 18, Nr. 12, 1990, Seiten 3587-3596, XP000652091 MORGENSTERN J P ET AL: "ADVANCED MAMMALIAN GENE TRANSFER: HIGH TITRE RETROVIRAL VECTORS WITH MULTIPLE DRUG SELECTION MARKERS AND A COMPLEMENTARY HELPER-FREE PACKAGING CELL LINE" *
PROGRESS IN NUCLEIC ACID RESEARCH AND MOLECULAR BIOLOGY, Bd. 38, 1990, Seiten 91-135, XP002030826 MCLACHLIN J R ET AL: "RETROVIRAL-MEDIATED GENE TRANSFER" *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6551587B2 (en) 1994-11-28 2003-04-22 Genetic Therapy, Inc. Vectors for tissue-specific replication
US6638762B1 (en) 1994-11-28 2003-10-28 Genetic Therapy, Inc. Tissue-vectors specific replication and gene expression
WO2002020060A1 (fr) * 2000-09-08 2002-03-14 Phogen Limited Agregats composes de la proteine vp22 et d'acides nucleiques et utilisation de tels agregats

Also Published As

Publication number Publication date
WO1997029201A3 (fr) 1997-10-02
DE19704979A1 (de) 1997-08-14

Similar Documents

Publication Publication Date Title
DE602004010799T2 (de) Synthetische bidirektionale promotoren und deren verwendungen
DE69936577T2 (de) Lentivirale Verpackungszellen
DE69535669T2 (de) Retrovirale vektoren mit verminderter rekombinationsrate
DE69636581T2 (de) Retrovirale Vektoren
DE69838758T2 (de) Verfahren und mittel zur herstellung von sicheren, rekombinanten lentivirusvektoren mit hohem titer
DE60028066T2 (de) Lentivirale vektoren für die herstellung von immunotherapeutischen zusammensetzungen
DE69829471T2 (de) Auf lentivirus basierende gentransfer-vektoren
EP1776460B2 (fr) Procede pour moduler l'expression genique par modification de la teneur en cpg
DE60023600T2 (de) Replizierende retrovirale Konstrukte, ihre Herstellung und Verwendungen für den Gentransfer
EP0778349A2 (fr) Construction génétique et son utilisation
CH684094A5 (de) Rekombinante Retroviren.
DE102008050860A1 (de) LCMV-GP-VSV-Pseudotypvektoren und tumorinfiltrierende Virenproduzentenzellen zur Therapie von Tumoren
US5670148A (en) Combined cellular and immunosuppressive therapies
EP1774008A2 (fr) Expression génique inductible
EP0777499B1 (fr) Vaccin vivant utilise pour traiter des maladies tumorales
EP1778295A1 (fr) Therapie genique de tumeurs solides par vecteurs retroviraux pseudotypes a arenavirus-glycoproteine
DE60038555T2 (de) Verpackungszelle
DE69627644T2 (de) Vektoren, die therapeutische gene fuer antimikrobielle peptide enthalten, zur verwendung in der gentherapie
DE10248141B4 (de) Nukleinsäuren und deren Verwendung für die Gentherapie
WO1997029201A2 (fr) Vecteur retroviral pour le transfert genique d'un antagoniste d'il6 dans des cellules souches hematopoietiques humaines
DE69930123T2 (de) Zielgerichtete integration in chromosomen mittels retroviraler vektoren
EP1173550B1 (fr) Procede pour reduire des immunoreactions specifiques
DE69635513T2 (de) Menschliche Pancreas Zelllinien: Entwicklungen und Anwendungen
US6485722B1 (en) Method for selective engraftment of drug-resistant hematopoietic stem cells
EP1220900A2 (fr) Vecteur de transfert de genes destine a la therapie de maladies auto-immunes et de maladies avec immunopathogenie

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A2

Designated state(s): JP US

AL Designated countries for regional patents

Kind code of ref document: A2

Designated state(s): AT BE CH DE DK ES FI FR GB GR IE IT LU MC NL PT SE

AK Designated states

Kind code of ref document: A3

Designated state(s): JP US

AL Designated countries for regional patents

Kind code of ref document: A3

Designated state(s): AT BE CH DE DK ES FI FR GB GR IE IT LU MC NL PT SE

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
121 Ep: the epo has been informed by wipo that ep was designated in this application
NENP Non-entry into the national phase

Ref country code: JP

Ref document number: 97528051

Format of ref document f/p: F

122 Ep: pct application non-entry in european phase