ES2550938T3 - Medicamentos y métodos para el tratamiento de mesotelioma - Google Patents
Medicamentos y métodos para el tratamiento de mesotelioma Download PDFInfo
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- ES2550938T3 ES2550938T3 ES08805222.0T ES08805222T ES2550938T3 ES 2550938 T3 ES2550938 T3 ES 2550938T3 ES 08805222 T ES08805222 T ES 08805222T ES 2550938 T3 ES2550938 T3 ES 2550938T3
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- 238000000034 method Methods 0.000 title abstract 2
- 206010027406 Mesothelioma Diseases 0.000 title description 3
- 229940079593 drug Drugs 0.000 title 1
- 239000003814 drug Substances 0.000 title 1
- 238000002483 medication Methods 0.000 title 1
- 210000004027 cell Anatomy 0.000 abstract description 14
- 210000004443 dendritic cell Anatomy 0.000 abstract description 5
- 208000015181 infectious disease Diseases 0.000 abstract description 2
- 239000013592 cell lysate Substances 0.000 abstract 2
- 208000006178 malignant mesothelioma Diseases 0.000 abstract 2
- 229960005486 vaccine Drugs 0.000 abstract 2
- 201000005505 Measles Diseases 0.000 abstract 1
- 230000002238 attenuated effect Effects 0.000 abstract 1
- 238000000338 in vitro Methods 0.000 abstract 1
- 210000001616 monocyte Anatomy 0.000 description 6
- 108010085238 Actins Proteins 0.000 description 3
- 102100037435 Antiviral innate immune response receptor RIG-I Human genes 0.000 description 3
- 101710127675 Antiviral innate immune response receptor RIG-I Proteins 0.000 description 3
- 230000001640 apoptogenic effect Effects 0.000 description 3
- 210000004881 tumor cell Anatomy 0.000 description 3
- 230000003612 virological effect Effects 0.000 description 3
- 102000007469 Actins Human genes 0.000 description 2
- 238000000684 flow cytometry Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 101100532034 Drosophila melanogaster RTase gene Proteins 0.000 description 1
- 229920001917 Ficoll Polymers 0.000 description 1
- 102000004457 Granulocyte-Macrophage Colony-Stimulating Factor Human genes 0.000 description 1
- 108010017213 Granulocyte-Macrophage Colony-Stimulating Factor Proteins 0.000 description 1
- 108010088729 HLA-A*02:01 antigen Proteins 0.000 description 1
- 102000006354 HLA-DR Antigens Human genes 0.000 description 1
- 108010058597 HLA-DR Antigens Proteins 0.000 description 1
- 101000946889 Homo sapiens Monocyte differentiation antigen CD14 Proteins 0.000 description 1
- 108090000978 Interleukin-4 Proteins 0.000 description 1
- 102100035877 Monocyte differentiation antigen CD14 Human genes 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 206010057249 Phagocytosis Diseases 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 230000000735 allogeneic effect Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 108091092328 cellular RNA Proteins 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 239000012228 culture supernatant Substances 0.000 description 1
- 238000000432 density-gradient centrifugation Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 230000008782 phagocytosis Effects 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 230000029812 viral genome replication Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/12—Viral antigens
- A61K39/155—Paramyxoviridae, e.g. parainfluenza virus
- A61K39/165—Mumps or measles virus
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/12—Viral antigens
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/46—Cellular immunotherapy
- A61K39/461—Cellular immunotherapy characterised by the cell type used
- A61K39/4611—T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/46—Cellular immunotherapy
- A61K39/461—Cellular immunotherapy characterised by the cell type used
- A61K39/4615—Dendritic cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/46—Cellular immunotherapy
- A61K39/462—Cellular immunotherapy characterized by the effect or the function of the cells
- A61K39/4622—Antigen presenting cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/46—Cellular immunotherapy
- A61K39/464—Cellular immunotherapy characterised by the antigen targeted or presented
- A61K39/4643—Vertebrate antigens
- A61K39/4644—Cancer antigens
- A61K39/464466—Adhesion molecules, e.g. NRCAM, EpCAM or cadherins
- A61K39/464468—Mesothelin [MSLN]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/46—Cellular immunotherapy
- A61K39/464—Cellular immunotherapy characterised by the antigen targeted or presented
- A61K39/4643—Vertebrate antigens
- A61K39/4644—Cancer antigens
- A61K39/464499—Undefined tumor antigens, e.g. tumor lysate or antigens targeted by cells isolated from tumor
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/46—Cellular immunotherapy
- A61K39/464—Cellular immunotherapy characterised by the antigen targeted or presented
- A61K39/464838—Viral antigens
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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- 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
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0634—Cells from the blood or the immune system
- C12N5/0639—Dendritic cells, e.g. Langherhans cells in the epidermis
- C12N5/064—Immunosuppressive dendritic cells
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- C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
- A61K2039/525—Virus
- A61K2039/5254—Virus avirulent or attenuated
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/58—Medicinal preparations containing antigens or antibodies raising an immune response against a target which is not the antigen used for immunisation
- A61K2039/585—Medicinal preparations containing antigens or antibodies raising an immune response against a target which is not the antigen used for immunisation wherein the target is cancer
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- C12N2760/00011—Details
- C12N2760/18011—Paramyxoviridae
- C12N2760/18411—Morbillivirus, e.g. Measles virus, canine distemper
- C12N2760/18432—Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent
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- C12N2760/00011—Details
- C12N2760/18011—Paramyxoviridae
- C12N2760/18411—Morbillivirus, e.g. Measles virus, canine distemper
- C12N2760/18434—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
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- Medicinal Chemistry (AREA)
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- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Mycology (AREA)
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- Engineering & Computer Science (AREA)
- Virology (AREA)
- Biomedical Technology (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Oncology (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Hematology (AREA)
- Communicable Diseases (AREA)
- Pulmonology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
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Abstract
Un método para preparar células dendríticas vacunales destinadas a tratar un mesotelioma maligno en un individuo, que comprende las siguientes etapas: - infección in vitro de células de mesotelioma maligno recogidas del individuo por una cepa de sarampión atenuada para producir un lisado celular; - poner en contacto células dendríticas con el lisado celular para producir células dendríticas vacunales.
Description
E08805222
22-10-2015
[0043] Con el fin de seguir la cinética del crecimiento vírico en cultivo de células M13 infectadas (MOI = 1,0), se realizaron RT-PCR específicas para receptores potenciales de ARNds vírico (Mda-5, TLR-3, RIG-I y PKR). Se usaron cebadores específicos para el gen β-actina como un control de experimento interno.
5 [0044] En resumen, las células M13 se incubaron con ligando de ácido poliinosínico:policitidílico (10 µg/ml) o VS (MOI = 1,0) y los gránulos celulares se recogieron en diferentes momentos. Después, todo el ARN celular se extrajo usando kits RNeasy (Qiagen, Courtaboeuf, Francia) de acuerdo con las instrucciones del fabricante, y se transcribieron a la inversa usando RTase (Invitrogen, Paisley, Reino Unido). Se usó el ADNc resultante como modelo
10 para la amplificación por PCR usando cebadores específicos para Mda-5, TLR-3, RIG-I, PKR, IFNβ y β-actina. Las secuencias de cebadores PCR se enumeran en la Tabla 1. Los productos PCR se visualizaron por electroforesis en gel de agarosa.
Tabla 1: Secuencias de cebador
- Cebador
- Secuencia Tamaño del fragmento (pb) SEQ ID NO:
- β-actina
- ATCTGGCACCACACCTTCTACAATGAGCTGCG directa 837 3
- CGTCATACTCCTGCTTGCTGATCCACATCTGC inversa
- 4
- TLR-3
- ATTGGGTCTGGGAACATTTCTCTTC directa 319 5
- GTGAGATTTAAACATTCCTCTTCGG inversa
- 6
- Mda-5
- GAGCAACTTCTTTCAACCAC directa 633 7
- GAACACCAGCATCTTCTCCA inversa
- 8
- RIG-I
- GAACGATTCCATCACTATCC directa 580 9
- GGCATCATTATATTTCCGCA inversa
- 10
- PKR
- CTTCTCAGCAGATACATCAG directa 689 11
- GTTACAAGTCCAAAGTCTCC inversa
- 12
15 [0045] Por lo tanto, puede mostrarse que se produjo un pico de replicación vírica entre 1 a 4 días después de la infección de las células M13 de mesotelioma. Además, también pudieron evidenciarse productos PCR correspondientes a receptores potenciales de ARNds vírico (Mda-5, TLR-3, RIG-I y PKR).
Captación eficiente de células de mesotelioma apoptóticas por CD inmaduros.
[0046] Después, se estudió la captación por células dendríticas (CD) de apocuerpos de células tumorales M13 25 infectadas por VS (72 horas) y se comparó con la de células tumorales M13 infectadas por UV (24 horas).
[0047] Las células dendríticas se obtuvieron a partir de monocitos generados de recogidas de leucoféresis de donantes sanos HLA-A0201 (EFS, Nantes, Francia), después de obtener el consentimiento informado. La fracción enriquecida con monocitos (>85 % de pureza) se separó en primer lugar por centrifugación por gradiente de 30 densidad Ficoll (PAA Laboratories, Les Mureaux, Francia). Después, los monocitos se enriquecieron por elutriación (centrifugación contracorriente) usando una centrífuga Beckman Avanti J20 equipada con un rotor JE5.0 y una cámara de elutriación de 40 ml. De forma habitual, la pureza de los monocitos elutriados estaba por encima del 80 %, como se evaluó por citometría de flujo en base a la detección del marcador CD14. Los monocitos se cultivaron en 2 x 108 células/ml con 500 IU/ml de GM-CSF y 200 IU/ml de IL-4 (Cell Genix Technology, Freiburg, Alemania).
35 Después, las células se dejaron diferenciar durante 6 días.
[0048] El día 6, las CD derivadas de monocitos se recogieron del sobrenadante de cultivo y se sembraron en cultivo para su carga posterior. Las CD inmaduras se incubaron con 2·108 células/ml de material apoptótico, derivado de células tumorales M13 alogénicas tratadas por UV o infectadas por VS, durante 24 horas más de cultivo 40 conjunto (relación 1:1). El análisis del rendimiento de la fagocitosis de las CD se evaluó tanto por citometría de flujo como microscopía láser confocal, como se ha descrito previamente (Massé y col. (2002) Cancer Research 32: 10501056). En resumen, las células M13 tratadas por UV o por VS se marcaron con colorante de tinción de membrana PKH-26, de acuerdo con el protocolo del fabricante (Sigma, St Quentin Fallavier, Francia). Después de 24 horas de cultivo conjunto, las CD se tiñeron con anticuerpos anti HLA-DR conjugados con FITC (Immunotech, Marseilles, 45 Francia). Después de los lavados con PBS, las células se cosecharon y se analizaron en un sistema FACSCalibur (BD Biosciences, Grenoble, Francia), o con un microscopio TCS NT (Leica Instruments, Heidelberg, Alemania). Las CD que habían ingerido células apoptóticas se identificaron como células doble positivo a HLA-DR/PKH-26 (figura
8
Claims (1)
-
imagen1
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP07291232A EP2058003A1 (en) | 2007-10-10 | 2007-10-10 | Medicaments and methods for treating mesothelioma |
| EP07291232 | 2007-10-10 | ||
| PCT/EP2008/063626 WO2009047331A1 (en) | 2007-10-10 | 2008-10-10 | Medicaments and methods for treating mesothelioma |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| ES2550938T3 true ES2550938T3 (es) | 2015-11-13 |
Family
ID=38935926
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| ES08805222.0T Active ES2550938T3 (es) | 2007-10-10 | 2008-10-10 | Medicamentos y métodos para el tratamiento de mesotelioma |
Country Status (7)
| Country | Link |
|---|---|
| US (2) | US9023643B2 (es) |
| EP (2) | EP2058003A1 (es) |
| CN (1) | CN101868249B (es) |
| CA (1) | CA2702186C (es) |
| DK (1) | DK2203183T3 (es) |
| ES (1) | ES2550938T3 (es) |
| WO (1) | WO2009047331A1 (es) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2432738A1 (en) * | 2003-02-26 | 2004-08-26 | Philippe Despres | New dengue and west nile viruses proteins and genes coding the foregoing, and their use in vaccinal, therapeutic and diagnostic applications |
| EP2058003A1 (en) | 2007-10-10 | 2009-05-13 | Institut National De La Sante Et De La Recherche Medicale (Inserm) | Medicaments and methods for treating mesothelioma |
| EP2759301A1 (en) | 2013-01-24 | 2014-07-30 | Institut Pasteur | Use of a genetically modified infectious measles virus with enhanced pro-apoptotic properties (MV-DeltaC virus) |
| CN107085095A (zh) * | 2017-03-02 | 2017-08-22 | 江苏华冠生物技术股份有限公司 | 用作酶联试剂盒包被抗原的麻疹病毒裂解液的制备方法 |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7118740B1 (en) * | 2000-09-22 | 2006-10-10 | Mayo Foundation For Medical Education And Research | Method for limiting the growth of cancer cells using an attenuated measles virus |
| EP2058003A1 (en) | 2007-10-10 | 2009-05-13 | Institut National De La Sante Et De La Recherche Medicale (Inserm) | Medicaments and methods for treating mesothelioma |
-
2007
- 2007-10-10 EP EP07291232A patent/EP2058003A1/en not_active Withdrawn
-
2008
- 2008-10-10 WO PCT/EP2008/063626 patent/WO2009047331A1/en active Application Filing
- 2008-10-10 EP EP08805222.0A patent/EP2203183B1/en active Active
- 2008-10-10 ES ES08805222.0T patent/ES2550938T3/es active Active
- 2008-10-10 DK DK08805222.0T patent/DK2203183T3/en active
- 2008-10-10 US US12/682,457 patent/US9023643B2/en active Active
- 2008-10-10 CA CA2702186A patent/CA2702186C/en active Active
- 2008-10-10 CN CN200880116771XA patent/CN101868249B/zh active Active
-
2015
- 2015-04-08 US US14/681,362 patent/US9636395B2/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| US9636395B2 (en) | 2017-05-02 |
| US20150209423A1 (en) | 2015-07-30 |
| EP2203183B1 (en) | 2015-08-19 |
| CA2702186A1 (en) | 2009-04-16 |
| WO2009047331A1 (en) | 2009-04-16 |
| US20100278872A1 (en) | 2010-11-04 |
| DK2203183T3 (en) | 2015-11-09 |
| US9023643B2 (en) | 2015-05-05 |
| CN101868249A (zh) | 2010-10-20 |
| CA2702186C (en) | 2019-05-28 |
| EP2203183A1 (en) | 2010-07-07 |
| CN101868249B (zh) | 2013-05-29 |
| EP2058003A1 (en) | 2009-05-13 |
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