EP0942961A1 - Procede de production de lymphocytes thymo-dependants tumoricides - Google Patents

Procede de production de lymphocytes thymo-dependants tumoricides

Info

Publication number
EP0942961A1
EP0942961A1 EP97948827A EP97948827A EP0942961A1 EP 0942961 A1 EP0942961 A1 EP 0942961A1 EP 97948827 A EP97948827 A EP 97948827A EP 97948827 A EP97948827 A EP 97948827A EP 0942961 A1 EP0942961 A1 EP 0942961A1
Authority
EP
European Patent Office
Prior art keywords
lymphocytes
cells
tumoricidal
tumor
antibody
Prior art date
Legal status (The legal status 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 status listed.)
Withdrawn
Application number
EP97948827A
Other languages
German (de)
English (en)
Inventor
Herbert Jungfer
Heinrich Barchet
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Roche Diagnostics GmbH
Original Assignee
Roche Diagnostics GmbH
Boehringer Mannheim GmbH
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 Roche Diagnostics GmbH, Boehringer Mannheim GmbH filed Critical Roche Diagnostics GmbH
Priority to EP97948827A priority Critical patent/EP0942961A1/fr
Publication of EP0942961A1 publication Critical patent/EP0942961A1/fr
Withdrawn legal-status Critical Current

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
    • 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
    • C12N5/0636—T lymphocytes
    • 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
    • A61K39/464—Cellular immunotherapy characterised by the antigen targeted or presented
    • A61K39/4643—Vertebrate antigens
    • A61K39/4644—Cancer antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K2035/124—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells the cells being hematopoietic, bone marrow derived or blood cells
    • 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
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/50—Cell markers; Cell surface determinants
    • C12N2501/515—CD3, T-cell receptor complex
    • 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
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/50—Cell markers; Cell surface determinants
    • C12N2501/599—Cell markers; Cell surface determinants with CD designations not provided for elsewhere

Definitions

  • the invention relates to a process for the activation, production and multiplication of tumorous T lymphocytes and a composition which is suitable as a medium for the production, activation and multiplication of tumorous T lymphocytes.
  • the cellular immune defense plays an important role in the elimination of pathologically altered body cells such as virus-infected cells or tumor cells.
  • cytotoxic T-lymphocytes recognize the changed body's own cells based on surface antigens.
  • surface antigens are usually protein fragments that are formed by the cells and are present on the cell surface bound to surface receptors of the so-called major histocompatibility complex (MHC) (Zinkernagel et al., Nature 248 (1974), 701-702 and Babbit et al., 1985, Nature 317: 359-361).
  • MHC major histocompatibility complex
  • LAK cells lymphokine-activated killer cells
  • the lymphocytes to be activated were also cultivated in the presence of autologous tumor cells (mixed lymphocyte tumor cultures, G. Fossati et al., International Journal of Cancer 42 (1988), 239-245; G. Degiovanni et al., Eur. J. Immunol. 18 (1988), 671-676; Wölfel et al., J. Exp. Med. 170 (1989), 797-810; Darrow et al., J. Immunol. 142 ( 1989), 3329-3335 and Notter et al., Int. J. Cancer 45 (1990), 834-841).
  • autologous tumor cells mixed lymphocyte tumor cultures, G. Fossati et al., International Journal of Cancer 42 (1988), 239-245; G. Degiovanni et al., Eur. J. Immunol. 18 (1988), 671-676; Wölfel et al., J. Exp. Med. 170 (1989), 797-810; Darrow
  • TLL tumor infiltrating lymphocytes
  • WO 94/23014 describes tumoricidal T lymphocytes and a process for their preparation. Thereafter, tumor tidal lymphocytes are produced by co-cultivating lymphocytes with a mammalian cell line of certain properties.
  • the object of the invention was to provide an improved and simplified method for the specific production of specifically tumoricidal T-lymphocytes, the Stimulation and proliferation of other T lymphocytes, and in particular of NK cells, is avoided
  • tumoricidal T-lymphocytes which cells of a chronic myelogenic leukemia cell line (K-562, ATCC CCL 243) do not destroy cytotoxically, but only inhibit their growth for a limited period of time, by cultivating lymphocytes which are lysosomotrophic be pretreated with anti-CD43 antibodies and isolation of the tumoricidal T lymphocytes.
  • lymphocytes white blood cells
  • sucker cell line a sucker cell line
  • Tumor lymphocytes in the sense of the invention, are T cells, as described in WO94 / 23014 and which recognize tumor cells in an antigen-unspecific manner and are killed by apoptosis induction and / or cytolysis. T tumor lymphocytes are characterized in that
  • no interleukin-2 is detectable in the culture supernatant of these tumorous T lymphocytes during the proliferation of these cells in the presence of the HB 654 or HB 617 cell line at a detection limit of 0.5 IU / ml
  • Tumor-killer T cells are not natural killer (NK) lymphocytes. Compared to tumor-infiltrating lymphocytes, they are better suited for therapeutic use due to their broader tumoricidal activity.
  • the tumoricidal cells T produced according to the method of the invention are T -Cells
  • Another distinguishing criterion between NK cells and tumoricidal T cells is that NK cells recognize the tumor cell line K-562 and destroy it cytotoxically, whereas tumoricidal T cells only inhibit the cell line K-562 for a limited time during growth
  • a time-limited inhibition in the sense of the invention is to be understood that the cells of the cell culture are not killed, but only inhibited in their proliferation, preferably completely inhibited in their proliferation. The inhibition of proliferation is reversible and can be reversed by separating the tumoricidal T cells and the cells of the cell line K-562.
  • a tumoricidal effect is not only to be understood as the killing (in particular lysis) of the tumor cell lines examined, but also an anti-proliferation effect.
  • This tumoricidal effect can e.g. are recognized by the cytotoxicity tests familiar to the person skilled in the art, e.g. the fact that the tumor cell lines, which are morphologically distinguishable from the tumor t-lymphocytes, disappear from the culture or at least remain in growth after prolonged cultivation with the tumor t-lymphocytes.
  • the tumoricidal T lymphocytes are preferably CD3 + and / or CD4 +.
  • Lymphocytes in the sense of the invention, are to be understood as preparations of white blood cells which can be obtained from whole blood, for example. Such methods are known to the person skilled in the art. For example, after a gradient centrifugation (Ficoll gradient), the fraction of mononuclear cells (PBMNC) containing the blood lymphocytes can be obtained.
  • PBMNCs obtained in this way have to be pretreated so that specifically tumoricidal T lymphocytes are formed.
  • the cells are pretreated using a lysosomotrophic substance, such as L-leucyl-leucine methyl ester or an analogous ester. Such substances are for example from PL.
  • the lysosomotrophic pretreatment maintains the fraction of surviving lymphocytes in which the progenitor cells are enriched with tumoricidal T-lymphocytes.
  • An anti-CD43 antibody is an antibody against the cell surface molecule CD43 (DeSmet, W. et al., In: Schlossman, SF, et al. [Eds.] Leukocyte Typing V, pp. 1706-1707, Oxford, United Kingdom: Oxford University Press, 1995).
  • CD43 is a sialoglycoprotein that is formed on cells of the hematopoietic system and - with the exception of dendritic cells and a subpopulation of B lymphocytes - is expressed on all cells of the immune system (Fukuda, M., Glycobiology 1 [1991], 347-356 ).
  • Such an antibody is preferably directed against the part of the CD43 molecule which is involved in the binding of CD43 to MHC class I molecules.
  • the antibody can be monoclonal or polyclonal and by immunization, synthesis or recombinant methods be made. Complete antibodies or antibody derivatives which mediate a characteristic binding can be used. However, an antibody or antibody derivative is preferably used which is capable of crosslinking CD43 molecules on cell surfaces in an analogous manner to an intact CD43 antibody.
  • one or more anti-CD2 antibodies are added to improve the yield of tumoricidal T lymphocytes, preferably a T cell stimulating pair (directed against two different epitopes of CD2) of anti-CD2 antibodies ( Schwarz, M., et al., J. Immunol. 154 (1995), 5813-5820; Moingeon, P., et al., Immunol. Rev. 111 (1989), 111-140).
  • the lymphocytes are cultured with the anti-CD43 antibody expediently in conventional culture media over several days (preferably 5-30 days).
  • the culture medium is preferably serum-free.
  • An autologous serum is very particularly preferably used for the cultivation if autologous tumoricidal T-lymphocytes are to be prepared.
  • the tumor tidal lymphocytes obtained in this way can, if necessary after cleaning, be administered to the patient as a therapeutic agent.
  • tumoral T-lymphocytes with a wide tumor activity can be obtained without HLA restriction.
  • a tumoricidal activity is understood to mean both a killing, in particular apoptosis-inducing and / or lysing, effect on the corresponding tumor cells and also a proliferation-inhibiting effect on these tumor cells.
  • Blood lymphocytes are preferably used as lymphocytes. However, it is also possible to use tumor infiltrating lymphocytes (TLL) as well as lymphocytes from spleen or lymph nodes. It is preferred to clean the lymphocyte preparation before use. When using blood lymphocytes, it is advisable to deplete the erythrocytes.
  • the fraction of the progenitor cells from tumoricidal T lymphocytes is enriched with the anti-CD43 antibody before cultivation.
  • monocytes, macrophages, NK cells, suppressor T cells and allogen-reactive, cytotoxic T cells and their precursor cells are eliminated lysosomotrophically, preferably according to Thiele and Lipsky (The Journal of Immunology, Vol. 136, No. 3 (1986) , Pp. 1038-1048), incubated with L-leucyl-L-leucine methyl ester.
  • lymphocytes preferably from the blood or from tumors of a donor.
  • lymphocytes according to known methods, e.g. isolated using a Ficoll density gradient centrifugation.
  • the lymphocytes left over after lysosomotrophic treatment are then cultured in a customary lymphocyte culture medium together with an anti-CD43 antibody.
  • the antibody concentration is between 1 ng / ml - 100 ⁇ g / ml, preferably 10 ⁇ g / ml culture medium.
  • Cultivation continues until the activation of tumoricidal T cells becomes apparent. This usually requires cultivation for around 8 days.
  • the cultivation according to the invention enables activation of tumoricidal T lymphocytes without growth or differentiation factors, such as e.g. Lymphokines, especially interleukin-2, must be added. This is of particular advantage for the therapeutic use of the tumoral T lymphocytes obtained, since such factors can trigger side effects in the therapeutic application.
  • human tumoricidal T-lymphocytes can be obtained without differentiation or growth factors, such as lymphokines, in particular interleukin-2, having to be added, which can lead to side effects during therapy.
  • Another object of the invention is therefore the use of the method according to the invention for the production of a therapeutic agent which is used in tumor therapy.
  • the tumoricidal T-lymphocytes produced according to the invention are washed according to methods familiar to the person skilled in the art (e.g. by centrifugation and resuspension of the pellet in physiological saline solution with repeated, e.g. triple repetition), if appropriate isolated and in a for suitable medium (e.g.
  • lymphocytes can also be activated in vivo by application of an anti-CD43 antibody to form tumoricidal T lymphocytes.
  • the antibody is taken up and applied in a medium suitable for the application, such as, for example, physiological saline.
  • the tumoricidal T lymphocytes can also be used for the elimination of tumor cells in a cell preparation ex vivo. It can preferably be used to isolate tumor cells from stem cell isolates (eg bone marrow stem cells) by culturing with tumoricidal T lymphocytes (purging). The stem cells cleaned in this way can be re-implanted in the patient after radiation or chemotherapy (autologous bone marrow transplantation).
  • stem cell isolates eg bone marrow stem cells
  • the stem cells cleaned in this way can be re-implanted in the patient after radiation or chemotherapy (autologous bone marrow transplantation).
  • Antibodies against the human CD43 molecule are described by DeSmet, W., et al. (In: Schlossman, S.F., et al. [Eds.] Leukocyte Typing V, pp. 1706-1707, Oxford, United Kingdom: Oxford University Press, 1995).
  • the anti-CD43 antibody designated as "6F5" is suitable.
  • an increase in the T-lymphocytes thus obtained is carried out.
  • the propagation is preferably carried out according to the method described in the international application PCT / EP97 / 01924.
  • the cell line HB 654 was deposited on March 24, 1993 with the German Collection for Cell Cultures and Microorganisms GmbH, Mascheroder Weg 1 b, D-38124 Braunschweig under the number DSM ACC 2122.
  • the cell line HB 617 was deposited on March 1st, 1994 with the German Collection for Cell Cultures and Microorganisms GmbH, Mascheroder Weg 1 b, D-38124 Braunschweig under the number DSM ACC 2166.
  • Mononuclear cells from the peripheral blood (PBMNC) of human donors are separated from the erythrocytes by gradient centrifugation (lymphocyte separation medium, Boehringer Manheim GmbH (BM)), washed twice with phosphate-buffered saline (Boehringer Mannheim GmbH (BM)) and at a density of approx 5 x 10 ⁇ cells / ml RPMI-1640 medium (Boehringer Mannheim GmbH (BM)) according to Thiele and Lipsky (The Journal of Immunology, Vol. 136, No. 3 (1986), pp. 1038-1048) at 250 ⁇ M leucyl-leucine methyl ester (Boehringer Mannheim GmbH (BM)) incubated for 20 minutes at room temperature.
  • BM peripheral blood
  • the surviving cells After washing twice with RPMI-1640 medium, the surviving cells at a density of approx. 2 x 10 ⁇ cells / ml in X-Vivo-20 medium (Bio-Whittacker), which the anti-CD43 antibody 6F5 in a Concentration of 10 ⁇ g / ml has been added, cultured at 37 ° C. in an 8% CO 2 atmosphere. On day 5 after the beginning of the culture, half of the medium is renewed and fresh anti-CD43 antibody 6F5 is added to a final concentration of 10 ⁇ g / ml. On day 10, after the culture has been set up, the cells are used for example 3.
  • the phenotyping of the cells on day 10 of the culture shows that> 95% of the cells are CD3 +. Cells with the CD 14 or CD 16 markers are not found.
  • lymphocytes are prepared from the blood of a human donor by gradient centrifugation and treatment with leucyl-leucine methyl ester.
  • the lymphocytes are suspended in Isove's modified DMEM (Gibco), to which 10% heat-inactivated (56 ° C / 30 minutes), autologous serum has been added, at a density of 1 x 10 ⁇ cells / ml and the anti-CD43 antibody 6F5 and the anti-CD2 antibodies TS2 / 18 and VIT13 (Schwarz, M., et al., J. Immunol. 154 (1995), 5813-5820) were added in a concentration of 10 ⁇ g / ml each. On day 5, half of the culture medium with the antibodies contained in it is renewed. The cell count on day 10 after the start of the culture showed that the lymphocytes had increased 2.2-fold.
  • the tumoricidal T-lymphocytes obtained according to Example 1 are added to cultures of human tumor lines (see Tables I and II). The tumoricidal effect on these tumor cells is monitored under the microscope. These different tumor cell lines are killed or their growth is inhibited with the tumoricidal T-lymphocytes according to the invention.
  • microorganism referred to under I has been received by this International Depository on (date of deletion) and an application has been made to convert this deletion into a deposit according to
  • microorganism referred to under I has been received by this International Depositary on (date of the deposit) and an application to convert this deposit into a deposit under the Budapest Treaty has been received on (date of receipt of the application for conversion).

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Microbiology (AREA)
  • Biomedical Technology (AREA)
  • General Health & Medical Sciences (AREA)
  • Cell Biology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • Veterinary Medicine (AREA)
  • Biotechnology (AREA)
  • Organic Chemistry (AREA)
  • Public Health (AREA)
  • Medicinal Chemistry (AREA)
  • Mycology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Biochemistry (AREA)
  • Hematology (AREA)
  • Oncology (AREA)
  • General Engineering & Computer Science (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

Le procédé selon l'invention, qui consiste à soumettre des lymphocytes à un prétraitement lysosomial, avec mise en incubation en présence d'anticorps anti-CD43, se prête à la fabrication de lymphocytes thymo-dépendants tumoricides.
EP97948827A 1996-10-30 1997-10-29 Procede de production de lymphocytes thymo-dependants tumoricides Withdrawn EP0942961A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
EP97948827A EP0942961A1 (fr) 1996-10-30 1997-10-29 Procede de production de lymphocytes thymo-dependants tumoricides

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
EP96117376 1996-10-30
EP96117376 1996-10-30
PCT/EP1997/005957 WO1998018907A1 (fr) 1996-10-30 1997-10-29 Procede de production de lymphocytes thymo-dependants tumoricides
EP97948827A EP0942961A1 (fr) 1996-10-30 1997-10-29 Procede de production de lymphocytes thymo-dependants tumoricides

Publications (1)

Publication Number Publication Date
EP0942961A1 true EP0942961A1 (fr) 1999-09-22

Family

ID=8223342

Family Applications (1)

Application Number Title Priority Date Filing Date
EP97948827A Withdrawn EP0942961A1 (fr) 1996-10-30 1997-10-29 Procede de production de lymphocytes thymo-dependants tumoricides

Country Status (5)

Country Link
EP (1) EP0942961A1 (fr)
JP (1) JP2000504230A (fr)
AU (1) AU6908898A (fr)
CA (1) CA2270117A1 (fr)
WO (1) WO1998018907A1 (fr)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20000058274A (ko) * 2000-02-24 2000-10-05 김규호 인터넷을 이용한 선구매 및 이익 분배 방법
CA2497628A1 (fr) * 2002-09-05 2004-03-18 Medimmune, Inc. Methodes de prevention ou de traitement de malignites cellulaires par administration d'antagonistes de cd2

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ZA91463B (en) * 1990-01-25 1992-09-30 Bristol Myers Squibb Co Method of activating cytolytic activity of lymphocytes using anti-cd28 antibody
DE4410136A1 (de) * 1993-03-31 1994-10-06 Boehringer Mannheim Gmbh Tumorizide T-Lymphozyten
WO1996015227A1 (fr) * 1994-11-14 1996-05-23 Novartis Ag Methodes destinees a induire la mort cellulaire de cellules hematopoietiques primitives et compositions a cet effet

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO9818907A1 *

Also Published As

Publication number Publication date
JP2000504230A (ja) 2000-04-11
AU6908898A (en) 1998-05-22
WO1998018907A1 (fr) 1998-05-07
CA2270117A1 (fr) 1998-05-07

Similar Documents

Publication Publication Date Title
DE60026561T2 (de) Suppressorzellen zur prävention und behandlung von immunantworten bei transplantationen
DE102017127984B4 (de) Verfahren für die Vermehrung und Aktivierung von γδ-T-Zellen
DE69834257T2 (de) Natürliche killerzelllinien und verfahren zu ihrer verwendung
DE60218302T2 (de) Verfahren zur gewinnung antigenspezifischer tr1-regualtorischer lymphozyten
DE69910362T2 (de) Anwendung der mesenchymalen stammzellen als immunsuppressiva
EP0885614B1 (fr) Procédé d'immunisation ex vivo utilisant des anticorps hétérologues intactes bispécifiques ou trispécifiques
DE3850745T2 (de) Verfahren zur Reinigung von rückständigen Tumorzellen in vitro mit lymphokinaktivierten zytotoxischen Zellen.
DE69729455T2 (de) Methode und zusammensetzungen zum erlangen reifer dendritischer zellen
US8603815B2 (en) CD4+ CD25− T cells and Tr1-like regulatory T cells
DE69213943T2 (de) Kurzzeitige stimulierung von lymphozyten mit anti-cd3 antikörpern um ihre in vivo aktivität zu steigern
EP0693120B1 (fr) Lymphocites t tumoricides
EP1331944B1 (fr) Utilisation d'anticorps monoclonaux specifiques a la molecule cd28 en vue de stimuler des cellules sanguines qui ne portent aucune molecule cd28
Werner et al. DX5+ NKT cells display phenotypical and functional differences between spleen and liver as well as NK1. 1-Balb/c and NK1. 1+ C57Bl/6 mice
DE60117167T2 (de) Verfahren zur herstellung von submikropartikel-suspensionen pharmazeutischer substanzen
EP0911388A2 (fr) Procédé pour la stimulation des lymphocytes T specifiques d'un antigène désiré
DE60130634T2 (de) Identifizierung von antigenen peptiden durch zytotoxische t-lymphozyten aktiviert von dendritischen zellhybriden
EP0942961A1 (fr) Procede de production de lymphocytes thymo-dependants tumoricides
DE69012105T2 (de) Proliferation von cd4-lymfocyten.
DE60016567T2 (de) Makrophagenhaltige zusammensetzung mit antiinfektiösen und hematopoietischen eigenschaften sowie deren herstellungsmethode
EP1381387A2 (fr) Utilisation de cellules mononucleaires du sang peripherique stimulees pour traiter des affections cancereuses
DE60038119T2 (de) Dendritische zellen, aktiviert in anwesenheit von glucocorticoiden hormonen, sind in der lage, die antigenspezifische t-zell-antwort zu unterdrücken
DE60033955T2 (de) Melanoma-impfstoff und seine herstellung und verwendung
DE60130206T2 (de) Zusammensetzungen und verfahren zur herstellung von antigen-präsentierenden zellen
EP2025747A1 (fr) Procédé d'amplification in vitro de lymphocytes T-régulateurs
EP0910624B1 (fr) Procede de production et de multiplication de lymphocytes

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 19990531

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): DE ES FR GB IT

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN

18D Application deemed to be withdrawn

Effective date: 20010503