EP0749487A1 - Reactif de liaison pour proteine de surface cellulaire et cellule effectrice - Google Patents

Reactif de liaison pour proteine de surface cellulaire et cellule effectrice

Info

Publication number
EP0749487A1
EP0749487A1 EP95911308A EP95911308A EP0749487A1 EP 0749487 A1 EP0749487 A1 EP 0749487A1 EP 95911308 A EP95911308 A EP 95911308A EP 95911308 A EP95911308 A EP 95911308A EP 0749487 A1 EP0749487 A1 EP 0749487A1
Authority
EP
European Patent Office
Prior art keywords
binding
binding reagent
reagent according
expression
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Withdrawn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
EP95911308A
Other languages
German (de)
English (en)
Inventor
Volker Schirrmacher
Khashayarsha Khazaie
Claudia Haas
Gerd Moldenhauer
Melvyn Little
Stefan Dübel
Frank Breitling
Sergey Kipriyanov
Stefanie Gotter
Hans-Jürgen RODE
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.)
Little Melvyn Prof Dr
RODE, HANS-JUERGEN, DR.
SCHIRRMACHER, VOLKER, PROF. DR.
Original Assignee
Deutsches Krebsforschungszentrum DKFZ
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 Deutsches Krebsforschungszentrum DKFZ filed Critical Deutsches Krebsforschungszentrum DKFZ
Publication of EP0749487A1 publication Critical patent/EP0749487A1/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • 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
    • 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
    • A61K39/0005—Vertebrate antigens
    • A61K39/0011—Cancer antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/08—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses
    • C07K16/10—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses from RNA viruses
    • C07K16/1027—Paramyxoviridae, e.g. respiratory syncytial virus
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
    • C07K16/2818—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against CD28 or CD152
    • 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
    • A61K2039/60—Medicinal preparations containing antigens or antibodies characteristics by the carrier linked to the antigen
    • A61K2039/6031—Proteins
    • A61K2039/6056—Antibodies

Definitions

  • the invention is therefore based on the object of providing a means by which the immunogenicity of cells can be increased.
  • the above compound is a B7 protein or a part thereof (cf. Nature 366 (1 993), 76; Science 262 (1 993), 909 "). It has proven to be particularly advantageous if such Part does not include the membrane-bound domain, and it is also advantageous if the compound is a lymphokine, interferon or interleukin.
  • effector cell encompasses any cell involved in an immune response. It is preferably a T cell.
  • costimulatory molecule includes any molecule of an effector cell that can be caused by binding or otherwise to stimulate the effector cell. Such a molecule can e.g. be a receptor.
  • a receptor e.g. be a receptor.
  • the receptors CD2-, CD3-, CD19-, CD20-, CD22-, CD26-, CD28- and CTLA-4 as well as HSA (Heat Stable Antigen) are particularly advantageous for the invention .
  • the receptor CD28 should be particularly emphasized.
  • binding reagents are preferred in which one or both binding components contain groups which form the formation of intermolecular disulfide bridges, i.e. of disulfide bridges between binding components of different binding reagents.
  • binding reagents that bind different, costimulatory molecules, coupled to a single cell surface protein. In many cases, this proves to be particularly beneficial for increasing the immunogenicity of cells.
  • Expression vector expression of the DNA, isolation of the expression product and its purification
  • a vaccine with inactivated cells is also provided, which is characterized in that one or more binding reagents are bound to a cell surface protein.
  • a vaccine preferably has Tumor cells. These can originate from tumors removed by surgery or from an established cell line.
  • binding reagents according to the invention which are to be attached to (tumor) cells, it is possible to transmit signals to molecules having a costimulatory effect, e.g. Receptors to transmit from effector cells. Effector cells thus not only receive the signal mediated by an antigen of (tumor) cells, but are also costimulated.
  • the binding reagents according to the invention are therefore particularly suitable for increasing the immunogenicity of cells, in particular tumor cells. They are a great improvement for active immunization.
  • FIG. 2 shows a schematic representation of the expression plasmid pOPE51- ⁇ CD28.
  • FIG. 3 shows a schematic representation of a binding reagent according to the invention.
  • anti HN anti-HN antibody
  • anti CD28 anti-CD28 antibody
  • S S: disulfide bridge.
  • the vector pOPE40 was used as the starting material (cf. Dübel et al., Gene 128 (1 993), 97). This contains a DNA which codes for the F v fragment (V H + V L ) of an anti-lysozyme antibody. The DNA for V H is linked to that for V L via a linker. The DNA above fragment is followed in the 3 'direction by a DNA which codes for an epitope of the monoclonal antibody 9E10 on the myc gene product.
  • Hybridoma cell line obtained using conventional PCR technology (cf. van Lier, R. et al. In Leucocyte Typing IV, Oxford University Press (1 989), 353).
  • the DNA of this fragment is referred to below as F v - ⁇ CD28-DNA.
  • the expression plasmid pOPE51 - ⁇ CD28 obtained is shown in FIG. 2.
  • E.coli JM 109 cells were transformed in the usual way with the expression plasmid pOPE51 - ⁇ HN and in 2 I LB medium at 30 ° C. up to an OD 600 of
  • Isopropyl- ⁇ -thiogalactoside IPTG was added up to a final concentration of 20 ⁇ m.
  • the cells were incubated at room temperature for 3 h and then collected by centrifugation (4 500 g, 4 ° C., 1 5 min).
  • the cells were suspended in 1/50 of the original volume in 0.1 M sodium acetate, pH 5.5, 10 mM EDTA, 1 mM phenylmethylsulfonyl fluoride (PMSF) at 0 °.
  • IPTG Isopropyl- ⁇ -thiogalactoside
  • Lysozyme was added to a final concentration of 1 mg / ml. After incubation on ice for 30 minutes with gentle shaking, soluble cell plasma proteins were removed by centrifugation (30,000 g, 4 ° C., 30 min). The cell pellets were resuspended in 1/40 of the original volume in 30 mM sodium phosphate, 0.3 M NaCl, 1 mM PMSF, pH 7.0 and lysed by sonication.
  • Imidazole was added to a final concentration of 30 mM.
  • the protein solution was placed on a chelating Sepharose Fast Flow column loaded with NiCl 2 and equilibrated with 6 M urea, 25 M Tris-HCl and 50 mM imidazole, pH 7.0.
  • the column was washed with five times the column volume of 6 M urea, 25 mM Tris-HCl, 50 mM imidazole, pH 7.0.
  • Bound F v - ⁇ HN fragments were eluted with 1.5 times the column volume of 6 M urea, 25 mM Tris-HCl, 250 mM imidazole, pH 7.0.
  • An IMAC was then carried out at room temperature.
  • the eluted protein was dialyzed at 4 ° C against 0.4 M L-arginine-HCl, 0.1 M Tris-HCl, 5 mM EDTA, pH 7.0, then using permeable Collodione-
  • Fat and connective tissue were removed from freshly operated tumor tissue in the usual way.
  • the tumor tissue was cut into small pieces and incubated with 40 ml of an enzyme cocktail (collagenase 0.32 mg / ml, DN-ase 0.535 mg / ml and hyaluronidase 0.535 mg / ml in HBSS) for 1 h at 37 ° C. with stirring.
  • the cell suspension obtained was poured off over a customary nylon mesh.
  • tissue residues were incubated a second time with the above enzyme cocktail (40 ml) and filtered. All suspensions were combined, made up to 50 ml with HBSS and centrifuged for 15 minutes at 1200 rpm. The supernatant was discarded and the pellet was resuspended in 10 ml of the above DN-ase solution and incubated at 37 ° C. for 10 min. The suspension was made up to 50 ml
  • the cell pellet obtained was resuspended in HBSS, 1 ⁇ 10 7 cells being taken up in 0.5 ml HBSS. Approx. 0.5 ml of the cell suspension obtained was placed in prepared freezing tubes. About 0.5 ml of double freezing medium on ice was added to these before the freezing tubes were stored at -70 ° C. overnight and then kept in liquid nitrogen.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Organic Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Virology (AREA)
  • Biochemistry (AREA)
  • Genetics & Genomics (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Animal Behavior & Ethology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Molecular Biology (AREA)
  • Biophysics (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Oncology (AREA)
  • Pulmonology (AREA)
  • Epidemiology (AREA)
  • Engineering & Computer Science (AREA)
  • Cell Biology (AREA)
  • Developmental Biology & Embryology (AREA)
  • Biotechnology (AREA)
  • Zoology (AREA)
  • Biomedical Technology (AREA)
  • Communicable Diseases (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

L'invention concerne un réactif de liaison qui se caractérise en ce qu'il comporte un premier constituant de liaison pour une protéine de surface cellulaire et un second constituant de liaison pour la molécule à effet costimulateur d'une cellule effectrice. L'invention concerne en outre un procédé de préparation dudit réactif de liaison, ainsi qu'un vaccin contenant ledit réactif de liaison.
EP95911308A 1994-03-07 1995-03-07 Reactif de liaison pour proteine de surface cellulaire et cellule effectrice Withdrawn EP0749487A1 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
DE4407538 1994-03-07
DE4407538A DE4407538C1 (de) 1994-03-07 1994-03-07 Bindungsreagens für Zell-Oberflächenprotein und Effektorzelle
PCT/EP1995/000843 WO1995024490A1 (fr) 1994-03-07 1995-03-07 Reactif de liaison pour proteine de surface cellulaire et cellule effectrice

Publications (1)

Publication Number Publication Date
EP0749487A1 true EP0749487A1 (fr) 1996-12-27

Family

ID=6512067

Family Applications (1)

Application Number Title Priority Date Filing Date
EP95911308A Withdrawn EP0749487A1 (fr) 1994-03-07 1995-03-07 Reactif de liaison pour proteine de surface cellulaire et cellule effectrice

Country Status (5)

Country Link
US (1) US5911987A (fr)
EP (1) EP0749487A1 (fr)
JP (1) JPH09510089A (fr)
DE (1) DE4407538C1 (fr)
WO (1) WO1995024490A1 (fr)

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB9607711D0 (en) * 1996-04-13 1996-06-19 Univ Sheffield T-cell dependent vaccine
DE19725586C2 (de) * 1997-06-17 1999-06-24 Gsf Forschungszentrum Umwelt Verfahren zur Herstellung von Zellpräparaten zur Immunisierung mittels heterologer intakter bispezifischer und/oder trispezifischer Antikörper
EP1092439A1 (fr) * 1999-10-13 2001-04-18 Thorsten Dr. Ahlert Activation de cellules T antigène spécifiques par des cellules dendritiques traitées par un virus/antigène
EP1275724A1 (fr) * 2001-07-10 2003-01-15 Volker Prof. Dr. Schirrmacher Réactifs de liaison entre des protéines de surface cellulaire et des cellules effectrices
CA2858359C (fr) * 2006-11-01 2018-04-03 Ventana Medical Systems, Inc. Haptenes, conjugues de haptene, compositions de haptene, procede de fabrication et utilisation

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5637481A (en) * 1993-02-01 1997-06-10 Bristol-Myers Squibb Company Expression vectors encoding bispecific fusion proteins and methods of producing biologically active bispecific fusion proteins in a mammalian cell
KR960701208A (ko) * 1993-03-16 1996-02-24 엔 엔 데이비스 바이러스 단백질에 의해 면역 반응을 자극하는 방법(stimulation of immune response by viral protein)

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO9524490A1 *

Also Published As

Publication number Publication date
DE4407538C1 (de) 1995-02-23
WO1995024490A1 (fr) 1995-09-14
US5911987A (en) 1999-06-15
JPH09510089A (ja) 1997-10-14

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