EP0409901A1 - Cellules de cytotoxine tueuses activees par la lymphokine - Google Patents

Cellules de cytotoxine tueuses activees par la lymphokine

Info

Publication number
EP0409901A1
EP0409901A1 EP89905571A EP89905571A EP0409901A1 EP 0409901 A1 EP0409901 A1 EP 0409901A1 EP 89905571 A EP89905571 A EP 89905571A EP 89905571 A EP89905571 A EP 89905571A EP 0409901 A1 EP0409901 A1 EP 0409901A1
Authority
EP
European Patent Office
Prior art keywords
lcc
cells
cytokine
pbmc
lak
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Withdrawn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
EP89905571A
Other languages
German (de)
English (en)
Other versions
EP0409901A4 (en
Inventor
Evan M. Hersh
Philip Scuderi, Jr.
William Grimes
Sydney E. Salmon
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.)
Research Corp Technologies Inc
Original Assignee
Research Corp Technologies Inc
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 Research Corp Technologies Inc filed Critical Research Corp Technologies Inc
Publication of EP0409901A1 publication Critical patent/EP0409901A1/fr
Publication of EP0409901A4 publication Critical patent/EP0409901A4/en
Withdrawn legal-status Critical Current

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Classifications

    • 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
    • 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
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/24—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against cytokines, lymphokines or interferons
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the present invention is directed to a unique group of cytokines produced by the stimulation of mammalian peripheral blood mononuclear cells with interleukin-2. More particularly, the present invention is directed to lymphokine activated killer cell cytotoxin (LCC), and to methods of production thereof, having unique cytotoxic and cytostatic activity against tumor cells.
  • LCC lymphokine activated killer cell cytotoxin
  • the present invention also contemplates a pharmaceutical composition comprising said cytokine and a pharmaceutically acceptable carrier useful in the treatment and prophylaxis of cancer and infectious diseases.
  • the present invention identifies a previously unknown cytokine associated with the LAK cell phenomenon, thereby providing a molecule with unique cytostatic and cytotoxic activity useful in the therapy of malignant and certain infection diseases.
  • the present invention is directed to a group of cytokines having unique cytotoxic and cytostatic activity against tumor cells.
  • Still another aspect of the present invention relates to antibodies specific to LCC useful in diagnostic assays for LCC.
  • LCC activity is measured in vitro using a microwell assay system.
  • tumor cells are placed in wells of 96 well plates in about 200 ul of complete medium and a volume of supernatant containing LCC is added.
  • Wells are evaluated for residual viable tumor cells after various periods of time.
  • LCC is found to be cytostatic in that it inhibits the proliferation of MCF-7 breast cancer cell line cells over 144 hours in culture.
  • LCC mediated activity against MCF-7 is not exhibited until after at least 24 hours of culture (Table 1).
  • the cytotoxic nature of LCC is evident against MCF-7 cells when added to a confluent monolayer thereof (Table 2). Accordingly, LCC can exhibit one or both of cytostatic and cytotoxic characteristics against tumor cells.
  • this assay detects the growth of clonogenic tumor stem cells. Inhibition in this assay in vitro correlates with inhibition of tumor growth in vivo in the patient donor of the tumor (Salmon and Trent, supra). LCC appears not to inhibit or kill normal cells. Thus, it neither inhibits adult or fetal fibroblasts (Table 7) nor does it inhibit normal lymphocytes responding to phytohemagglutinin (PHA) or IL-2 stimulation (Table 8). Accordingly, the LCC of the present invention has a wide spectrum of activity against cancer and tumor cells and a normal and neoplastic cells. Hereinafter, the word tumor will be used to define such cell types including infectious diseases causing transformation of said cell types. Hence, as defined herein, tumor or tumor cells is meant to encompass all LCC-sensitive cells and LCC-target cells.
  • LCC is distinct from (in addition to the above) IFN- ⁇ , IL-1, leukoregulin, NKCF, perforin, cytolysin, serine esterase, lysozomal arylsulfatase, TGF ⁇ 1, and oncostatin M.
  • LCC has been partially purified by FPLC, the supernatant of PBMC not stimulated with IL-2 ( Figure 1A) lacks a peak (number 5) present in the supernatant FPLC fractionation of IL-2 stimulated PBMC ( Figure 1B). This fraction also contains the LCC activity. Crude LCC was further purified and characterized. LCC is tightly associated with serum albumin. Passage of crude LCC over a Q-Sepharose (Pharmacia) column yields two fractions LCC-1 and LCC-2. LCC-1 is the flow through fraction and has potent cytotoxic activity while being devoid of albumin. LCC-2 is eluted from the column by treatment with a solution having a high salt concentration.
  • the salt solution consists of an amount and type of salt having an anion effective to displace LCC from the Q-Sepharose resin to yield the fraction designated LCC-2.
  • the salt solution is about 0.5 M to about 1.5 M NaCl, and most preferably is 1.0 M NaCl.
  • LCC-1 retains its cytotoxic activity after removal of TNF ⁇ or IFN ⁇ and is useful for the final purification of LCC. TNF ⁇ - and IFN ⁇ -free LCC-1 and any purified fraction demonstrating LCC activity is useful in the generation antibodies, especially monoclonal antibodies.
  • the cytokine LCC is contemplated herein to be useful in the regression and pallation of some cancer, tumor and infectious diseases in mammals and preferably in humans. Such diseases where LCC is considered most useful are those where cell cytostasis or cytotoxicity (cell killing) of target cells is desired.
  • a target cell is used herein to describe all cells (e.g. tumor and cancer) associated with said disease states.
  • the subject invention contemplates a method for inducing regression or inhibition of growth of said target cells in mammals (i.e. cancers and tumors) by administering a pharmaceutical composition containing an effective amount of said LCC.
  • a method for inducing regression -or inhibition of growth of cancer in a mammal is contemplated in which a nucleic acid molecule encoding the LCC contemplated herein is introduced into an affected (i.e., cancerous or transformed) cell in such a manner that said nucleic acid molecule is expressed intracellularly but extrachromosomally of said cell or following integration into the genome of said cell.
  • the nucleic acid molecule is carried to said affected cell and transferred into said cell by a second nucleic acid molecule (e.g., various viruses).
  • the first nucleic acid molecule is manipulated such that it contains the appropriate signals for expression. That is, in accordance with the present invention, a method of inducing regression or inhibition of growth of cancer in a mammal is contemplated comprising administering a first nucleic acid molecule encoding LCC, said nucleic acid being contained in a pharmacologically acceptable second nucleic acid carrier molecule such that said first nucleic acid enters a target cell and is either maintained extrachromosomally or integrates into the genome of said target all in such a manner that said first nucleic acid is expressed so as to produce an effective amount of LCC.
  • the active ingredients of the pharmaceutical compositions comprising LCC are contemplated to exhibit excellent and effective therapeutic activity, for example, in the treatment of some cancers and tumors or infectious diseases.
  • the active ingredients of the therapeutic compositions including LCC exhibit antitumor activity when administered in amounts from about 0.5 ug to about 2000 ⁇ q per kilogram of body weight per day.
  • This dosage regimen may be adjusted to provide the optimum therapeutic response. For example, several divided doses may be administered daily or the dose may be proportionally reduced as indicated by the exigencies of the therapeutic situation.
  • a decided practical advantage is that the active compound may be administered in a convenient manner such as by the oral, intraveneous (where water soluble), intramuscular or subcutaneous routes.
  • the active compounds may also be administered parenterally or intraperitoneally.
  • Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.
  • the pharmaceutical forms suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion.
  • the form must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi.
  • the carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils.
  • the proper fluidity can be maintained, for example, by the use of a coating such as licithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants.
  • Thepreventions of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin.
  • pharmaceutically acceptable carrier includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like.
  • the use of such media and agents for pharmaceutical active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredient, its use in the therapeutic compositions is contemplated. Supplementary active ingredients can also be incorporated into the compositions.
  • Dosage unit form refers to physically discrete units suited as unitary dosages for the mammalian subjects to be treated; each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier.
  • the specification for the novel dosage unit forms of the invention are dictated by and directly dependent on (a) the unique characteristics of the active material and the particular therapeutic effect to be achieved, and (b) the limitations inherent in the art of compounding such an active material for the treatment of disease in living subjects having a diseased condition in which bodily health is impaired as herein disclosed in detail.
  • the principal active ingredient is compounded for convenient and effective administration in effective amounts with a suitable pharmaceutically acceptable carrier in dosage unit form as hereinbefore disclosed.
  • a unit dosage form can, for example, contain the principal active compound in amounts ranging from 0.5 ⁇ g to about 2000 ⁇ g . Expressed in proportions, the active compound is generally present in from about 0.5 to about 2000 ⁇ g/ml of carrier.
  • the dosages are determined by reference to the usual dose and manner of administration of the said ingredients.
  • the present invention also relates to antibodies to LCC.
  • Such antibodies are contemplated to be useful indeveloping detection assays (immunoassays) for said LCC, especially during the monitoring of a therapeutic regimen and in the purification of LCC.
  • the antibodies may be monoclonal or polyclonal.
  • the present invention further contemplates use of these second antibodies in detection assays and, for example, in monitoring the effect of an administered pharmaceutical preparation.
  • an antibody to LCC encompasses antibodies to LCC, or part thereof, and to any associated molecules (e.g., glycosylated regions, lipid regions, carrier molecules, and the like).
  • the LCC, or parts thereof, considered herein are purified, as for example using FPLC, then utilized in antibody production.
  • Both polyclonal and monoclonal antibodies are obtainable by immunization with the polypeptides, and either type is utilizable for immunoassays.
  • the methods of obtaining both types of sera are well known in the art.
  • Polyclonal sera are less preferred but are relatively easily prepared by injection of a suitable laboratory animal with an effective amount of the purified polypeptide, or part thereof, collecting serum from the animal, and isolating specific sera by any of the known immunoadsorbent techniques.
  • antibodies produced by this method are utilizable in virtually any type of immunoassay, they are generally less favored because of the potential heterogeneity of the product.
  • the use of monoclonal antibodies in the present immunoassay is particularly preferred because of the ability to produce them in large quantities and the homogeneity of the product.
  • the preparation of hybridoma cell lines for monoclonal antibody production derived by fusing an immortal cell line and lymphocytes sensitized against the immunogenic preparation can be done by techniques which are well known to those who are skilled in the art. (See, for example, Douillard, J. Y. and Hoffman, T., "Basic Facts About Hybridomas", in Compendium of Immunology, Vol. II, L. Schwartz (Ed.) (1981); Kohler, G. and Milstein, C, Nature 256: 495-497 (1975); European Journal of Immunology, Vol. 6, pp.
  • the choice of animal is dependent on the availability of appropriate immortal lines capable of fusing with lymphocytes thereof.
  • Mouse and rat have been the animals of choice in hybridoma technology and are preferably used. Humans can also be utilized as sources for sensitized lymphocytes if appropriate immortalized human (or nonhuman) cell lines are available.
  • the animal of choice may be injected with from about 1 mg to about 20 mg of the purified LCC, or part thereof. Usually the injecting material is emulsified in Freund's complete adjuvant. Boosting injections may also be required. The detection of antibody production can be carried out by testing the antisera with appropriately labeled antigen.
  • a number of cell lines suitable for fusion have been developed, and the choice of any particular line for hybridization protocols is directed by any one of a number of criteria such as speed, uniformity of growth characteristics, deficiency of its metabolism for a component of the growth medium, and potential for good fusion frequency.
  • Cell fusion can be induced either by virus, such as Epstein-Barr or Sendai virus, or polyethylene glycol.
  • virus such as Epstein-Barr or Sendai virus
  • polyethylene glycol Polyethylene glycol (PEG) is the most efficacious agent for the fusion of mammalian somatic cells. PEG itself may be toxic for cells, and various concentrations should be tested for effects on viability before attempting fusion.
  • the molecular weight range of PEG may be varied from 1,000 to 6,000. It gives best results when diluted to from about 20% to about 70% (w/w) in saline or serum-free medium. Exposure to PEG at 37°C for about 30 seconds is preferred in the present case, utilizing murine cells.
  • an unlabeled antibody is immobilized in a .solid substrate and the sample to be tested brought into contact with the bound molecule.
  • a second antibody labeled with a reporter molecule capable of producing a detectable signal is then added and incubated, allowing time sufficient for the formation of a ternary complex of antibody-labeled antibody. Any unreacted material is washed away, and the presence of the antigen is determined by observation of a signal produced by the reporter molecule.
  • the results may either be qualitative, by simple observation of the visible signal, or may be quantitated by comparing with a control sample containing known amounts of hapten.
  • Variations on the forward assay include a simultaneous assay, in which both sample and labeled antibody are added simultaneously to the bound antibody, or a reverse assay in which the labeled antibody and sample to be tested are first combined, incubated and then added to the unlabeled surface bound antibody.
  • a first antibody having specificity for LCC, or part thereof. contemplated in this invention is either covalently or passively bound to a solid surface.
  • the solid surface is typically glass or a polymer, the most commonly used polymers being cellulose, polyacrylamide, nylon, polystyrene, polyvinyl chloride or polypropylene.
  • the solid supports may be in the form of tubes, beads, discs or microplates, or any other surface suitable for conducting an immunoassay.
  • the binding processes are well-known in the art and generallyconsist of cross-linking, covalently binding or physically adsorbing the molecule to the insoluble carrier. Following binding, the polymer-antibody complex is washed in preparation for the test sample.
  • reporter molecule a molecule which, by its chemical nature, provides an analytically identifiable signal which allows the detection of antigen-bound antibody. Detection may be either qualitative or quantitative.
  • reporter molecules in this type of assay are either enzymes, fluorophores or radionuclide containing molecules (i.e., radioisotopes).
  • an enzyme is conjugated to the second antibody, generally be means of glutaraldehyde or periodate.
  • Commonly used enzymes include horseradish peroxidase, glucose oxidase, B-galactosidase and alkaline phosphates, among other.
  • the substrates to be used with the specific enzymes are generally chosen for the production, upon hydrolysis by the corresponding enzyme, of a detectable color change.
  • p-nitrophenyl phosphate is suitable for use with alkaline phosphatase conjugates; for peroxidase conjugates, 1, 2-phenylenediamine, 5-aminosalicyclic acid, or tolidine are commonly used.
  • fluorogenic substrates which yield a fluorescent product rather than the chromogenic substrates noted above.
  • the enzyme-labeled antibody is added to the first antibody-hapten complex, allowed to bind, and then the excess reagent is washed away. A solution containing the appropriate substrate is then added to the ternary complex of antibody-antigen-antibody. The substrate will react with the enzyme linked to the second antibody, giving a qualitative visual signal, which may be further quantitated, usually spectrophotometrically, to give an indication of the amount of hapten which was present in the sample.
  • a recombinant nucleic acid molecule said molecule defined herein to be DNA or RNA, encoding LCC or part thereof.
  • the recombinant nucleic acid molecule is complementary DNA (cDNA). It is considered within the scope of the present invention to include the cDNA molecule encoding mammalian LCC, but preferably human LCC, or to regions or parts thereof including any base deletion, insertion or substitution or any other alteration with respect to nucleotide sequence or chemical composition (e.g. methylation and glycosylation). LCC encoded by cDNA is referred to herein as recombinant LCC.
  • the scope of the present invention therefore, encompasses recombinant LCC and the cDNA and mRNA encoding LCC including any host carrying and optionally expressing recombinant LCC cDNA and mRNA.
  • LCC The TNF resistant variant of L929 cells which is resistant to both TNF- ⁇ and TNF- ⁇ is sensitive to LCC (Table 16).
  • LCC is distinct from (in addition to the above) IFN- ⁇ , IL-1, leukoregulin, NKCF, perforin, cytolysin, serine esterase, lysozomal arylsulfatase, TGF ⁇ 1, and oncostatin M.
  • Some of these relate to the resistance of K562 cells to LCC (Table 17) which is known to be sensitive to NKCF, Leukoregulin, perforin, cytolysin, etc.
  • Cytokine Direct Cytokine Resistant Cytokine Sensitive Assay Line is LCC Line is LCC Sensitive Resistant
  • TGF ⁇ 1 Inhibits PHA stimulated blasts Oncostatin M Does not inhibit L292. MW 28,000 produced by purified T cells over 3-28d.
  • FIG. 1 is a graphical representation of LCC activity in FPLC fractions.
  • Figure 1A represents fractions derived from supernatant of PBMC incubated in the absence of IL-2. No peak is seen at number 5. In difference, supernatant from IL-2 incubated cells has a peak at number 5. This also contains LCC activity.
  • crude LCC (LAK cell supernatent made in AIM 5 medium) was concentrated 10-fold to 100 ml using an Amicon Spiral Centrifuge (MW cut-off 10,000) and exchanged with 1000 ml of 50 mM MES buffer (pH 6.0) using the same spiral centrifuge technique until the ultrafiltrate reached pH 6.0.
  • the concentrated LCC was loaded onto a Q-Sepharose column (50 ml bed volume, equilibrated with 50 mM MES, pH 6.0) with a peristaltic pump operating at a flow rate of 3 ml/min.
  • LCC-1 has potent cytotoxic activity, is devoid of albumin but contains transferrin.
  • the proteins bound to the Q-Sepharose column were eluted with 1 M NaCl and exchanged into RPMI medium by Centriprep 30 ultrafiltration. This fraction is designated LCC-2, contains almost all the serum albumin and has potent cytotoxic activity.
  • Crude LCC, LCC-1 and LCC-2 contain small but measurable amounts of known cytokines including TNF and IFN ⁇ . These activities are measured in crude LCC, LCC-1 and LCC-2 by ELISA. These various fractions were subjected to affinity purification to remove contaminants using columns coated with anti-TNF ⁇ or anti-IFN ⁇ antibodies. The results shown in Table 20 indicated that albumin-free LCC-1 retains much of its cytotoxic activity after removal of TNF ⁇ or IFN ⁇ while albumin-containing LCC-2 loses much of its activity with the same treatment. After passage through both columns, crude LCC contained no measurable TNF ⁇ and only 22.5 u/ml IFN ⁇ whereas LCC-1 contained no TNF ⁇ or IFN ⁇ . An amount of IFN ⁇ of 22.5 u/ml (or even 1000 u/ml) has no effect on MCF-7 cells.

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  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Biophysics (AREA)
  • Biochemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Molecular Biology (AREA)
  • Genetics & Genomics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Veterinary Medicine (AREA)
  • Immunology (AREA)
  • Public Health (AREA)
  • Pharmacology & Pharmacy (AREA)
  • General Chemical & Material Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Toxicology (AREA)
  • Zoology (AREA)
  • Gastroenterology & Hepatology (AREA)
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Abstract

Cette invention concerne un groupe unique de cytokines produit par la stimulation de cellules mononucléaires de sang périphérique de mammifère avec l'interleukine-2. Plus particulièrement, cette invention concerne des cellules de cytotoxine tueuses activées par la lymphokine et leurs procédés de production, ces cellules ayant une activité cytotoxique et cytostatique unique contre des cellules cancéreuses. L'invention concerne également une composition pharmaceutique comportant ladite cytokine et un porteur acceptable du point de vue pharmaceutique qui est utile dans le traitement et la prophylaxie du cancer et des maladies infectieuses.
EP19890905571 1988-04-15 1989-04-05 Lak cell cytotoxin Withdrawn EP0409901A4 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US18187088A 1988-04-15 1988-04-15
US181870 1988-04-15

Publications (2)

Publication Number Publication Date
EP0409901A1 true EP0409901A1 (fr) 1991-01-30
EP0409901A4 EP0409901A4 (en) 1991-10-30

Family

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Family Applications (1)

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EP19890905571 Withdrawn EP0409901A4 (en) 1988-04-15 1989-04-05 Lak cell cytotoxin

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EP (1) EP0409901A4 (fr)
JP (1) JPH03503844A (fr)
WO (1) WO1989009831A1 (fr)

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1990004412A1 (fr) * 1988-10-27 1990-05-03 Regents Of The University Of Minnesota Immunoadjuvants liposomiques contenant de l'il-2
US5725855A (en) * 1991-04-05 1998-03-10 The United States Of America As Represented By The Department Of Health And Human Services Method of treating tumors with CD8+ -depleted or CD4+ T cell subpopulations
AU663711B2 (en) * 1991-04-05 1995-10-19 Regents Of The University Of Minnesota Method of enhancing the immunotherapeutic activity of immune cells by depletion/positive selection of cell subsets
IE922233A1 (en) * 1991-07-10 1993-01-13 Augusto C Ochoa Short-term anti-cd3 stimulation of lymphocytes to increase¹their in vivo activity
EP0624095B2 (fr) * 1991-12-31 2005-07-20 Zymogenetics, Inc. Procedes et compositions pour reduire la perte de sang

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0186833B1 (fr) * 1984-12-20 1992-08-19 Yeda Research And Development Company Limited Anticorps monoclonal contre une cytotoxine, hybridome le produisant et procédé pour la préparation de la cytotoxine

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4530901A (en) * 1980-01-08 1985-07-23 Biogen N.V. Recombinant DNA molecules and their use in producing human interferon-like polypeptides

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0186833B1 (fr) * 1984-12-20 1992-08-19 Yeda Research And Development Company Limited Anticorps monoclonal contre une cytotoxine, hybridome le produisant et procédé pour la préparation de la cytotoxine

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
CANCER RESEARCH, vol. 48, no. 16, 15th August 1988, pages 4687-4688, Philadelphia, USA; D.M. LOWREY et al.: "Isolation and characterization of cytotoxic granules from human lymphokine (interleukin 2) activated killer cells" *
PROC. AM. ASSOC. CANCER. RES. 79TH ANNU. MEET., vol. 29, March 1988, page 399, abstract no. 1588; E.M. HERSH et al.: "A soluble antitumor factor produced during interleukin-2 (IL-2) induced generation of lymphokine activated killer (LAK) cells. Evidence for a dual mechanism of lak-cell action" *
See also references of WO8909831A1 *

Also Published As

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EP0409901A4 (en) 1991-10-30
WO1989009831A1 (fr) 1989-10-19
JPH03503844A (ja) 1991-08-29

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