CN1142186A - Use of IL-10 to stimulate peripheral blood mononuclear cell cytolytic activity - Google Patents
Use of IL-10 to stimulate peripheral blood mononuclear cell cytolytic activity Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/19—Cytokines; Lymphokines; Interferons
- A61K38/20—Interleukins [IL]
- A61K38/2013—IL-2
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- A—HUMAN NECESSITIES
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- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
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- A61K38/19—Cytokines; Lymphokines; Interferons
- A61K38/20—Interleukins [IL]
- A61K38/2066—IL-10
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- A—HUMAN NECESSITIES
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/19—Cytokines; Lymphokines; Interferons
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- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
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- Chemical Kinetics & Catalysis (AREA)
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Abstract
The invention relates to a usage of IL-10. The alone usage of IL-10 or combination of IL-10 and IL-2 or Alpha-IFN can excite cytolysis activity of peripheral blood monocyte phenotype to cure cancer.
Description
The present invention relates to the use of IL-10 INTERLEUKIN-10, IL-10 INTERLEUKIN-10 (IL-10) is CSIF (CSIF) is used for tumor disease (cancer) by the cell lysis activity that excites peripheral blood lymphocytes (PBMC) a adoptive immunotherapy in form.
Background of invention
The immunotherapy of oncotherapy is based on such notion: cancerous cell since certain also not clear former thus escaped body can be for the defense reaction of the cell of distortion or external source and molecule and this defense reaction by the method deexcitation for the treatment of to kill or the growth of anticancer, for example Klein once discussed this problem (Immunology, Wiley-Interscience, 1982 pp.623-648).Immune effector can suppress the nearest observation of growth of tumor directly or indirectly as a result, makes people produce interest again to the method for this treatment tumor.[Heberman，Concepts?Immunopath.1：96(1985)(natural?killercells?resist?tumor?growth)；Rosenberg?et?al.，Ann.Rev.Immunol.4：681(1988)(use?of?IL-2-activated?killer?cellls?to?treat?cancerRalf?et?al.，J.Exp.Med.167：712(1988)(tumoricidal?activity?ofmacrophages?stimulated?by?lymphokines)；Tepper?et?al.，Cell?57：503(1989)(IL-4?has?tumoricidal?activity)；M.Cohen，“Lymphokines?and?Tumor?Immunity”，pp.237-253，in?S.Cohened.，Lymphokines?and?the?Immune?Response(CRC?Press，Boca?Raton，1990)]。
The immunologic responsiveness of tumor is subjected to the regulation and control of polytype cell but also comprises the effect of cytokines of T cell and cells of monocytic origin.A kind of clinical immunotherapy method that is expected to is exactly a what is called " adoptive immunity therapy " of using the activated killer cell of IL-2.[Rosenberg，Supra?Rosenberg，Sci.Am，pp.62-69(May?1990)]。Although IL-2 share very effective when controlling some malignant tumor (for example renal cell carcinoma) separately or with traditional chemotherapeutics, but the disadvantageous malicious pair that use brought that is accompanied by the IL-2 of effective dose is appointed for example vascular lamina seepage and edema, and the danger that makes some people propose this Therapeutic Method has surpassed the benefit of itself.[Cotran?et?al.，J?Immunol.139：1882(1987)；Edwards?et?al.，Cancer?Res.52：3425(1992)]。
Human vascular endothelial is responsive especially to the toxicity of IL-2, and showing as vascular permeability increases (being the vascular leakage syndrome) and edema.Cause that a kind of IL-2 of being activated T cells and the adhesive attraction of neutrophilic granulocyte on the monolayer of blood vessel endothelium in the possible factor of this pathological reaction strengthen, this point as in the past the external work of doing certified [Edwars et al., Supra; Damle et al., 138:1779 (1987)].
Another kind is the adoptive transfer of immunocyte to the method for the immunization therapy of tumor disease.The definition of adoptive immunotherapy is: shift or transplant the cell with anti-tumor activity that the active immne medicament for example can directly or indirectly mediate antitumor action in the host of carrying tumor.The adoptive immunity therapy is the method for a very attractive treatment tumor disease.Should be noted that owing to changing the host over to intravital be activated immuning agent, therefore do not need host's immunocompetence completely.Therefore, the immunosuppressant that causes owing to the trouble tumor usually is not a major obstacle in this immunotherapy.Because host's immunocompetence do not require, and in fact this situation may be useful to the adoptive transfer of immunocyte, so the use that can combine with other Therapeutic Method such as chemotherapy and radiation at an easy rate of adoptive immunity therapy.And different with other Therapeutic Method be that this therapy may not can produce immunosuppressive action.
Tumour patient certainly will cause immunologic injury in chemotherapy or radiotherapy, and the supply that will often make activate immunity react effective effector pericellular blood monocyte (PBMCs) reduces.At low effector cell: under the target cell ratio condition with regard to the Therapeutic Method of expression effect thereby will be particularly advantageous to some patients like this.
The immunoreactivity of tumor is subjected to the regulation and control of polytype cell and comprises the effect of cytokines of T cell and cells of monocytic origin.Use use [the Rosenkey et al. of the adoptive immunity therapy of the human cell factor of recombinating along with IL-2, J.Immol 138:1779 (1985)] related to peripheral blood lymphocytes (PBMCs) external activation [Philips et al., J.Clin.Oncd.5:1933:(1987); Perussia, Carr, Opion Immunol.3:49:(1991)] extracorporeal treatment of peripheral blood lymphocytes produces activated killer cell of activatory lymphokine (LAK) and activated natural killer cell (NK), and the two all has cell lysis activity to various tumor cell.
Human interleukin-5 (IL-5) [the Nagasawa et al. of existing report reorganization, Cell.Immunol.133:317 (1991)], interleukin-7 (IL-7) [Stotter and Lotze, Arch.Surgery126:1525 (1991)] and il-1 2 (IL-12) [Gately et al., J.Immunol.147:874 (1991)] can activate human peripheral blood mononuclear cell's cell lysis activity.The initial report of IL-10 INTERLEUKIN-10 (IL-10) is to be secreted as a kind of cytokine inhibitory factor by special t helper cell subclass in mice, the differentiation [Chen and Zlotnik, J.Immunol 147:528 (1991)] of scalable mouse cell poison T cell.Have the supernatant of the COS cell recovery of people IL-10 cDNA to hatch the human peripheral blood mononuclear cell but also find to use from transfection, the cell through hatching is external solubilized tumor target cell.To the IL-10 dissolving potentiality enhanced T cell that reacts, through being accredited as a CD56+ phenotype, explanation is NKT (NK) cell.
In some cases, IL-4 is to having produced opposite effect by the inductive LAK activity of IL-2 institute.[Nagler?et?al.，J.Immunol.141：2349(1988)]。For example, if the human peripheral blood mononuclear cell is hatched with IL-2 and IL-4 simultaneously, the target cell lysis of LAK sensitivity is significantly reduced [Spits et al., J.Immunol.141:29 (1988)].If but PBMCs had the culture medium of IL-2 to cultivate in advance 3 days with mending before adding IL-4, the lytic activity of cell all is [Spits et al., the Supra] that increases.If in initial mixtures incubated, comprise alpha-interferon (α-IFN) or tumor necrosis factor-alpha (TNF-α), then IL-4 can weaken [Swisher et al., Cell.Immunol.128:450 (1990)] to I-2 to the blocking effect of the cell lysis activity that drives.
Kedar etc. [Cancer Immunol.Immunother.35:63 (1992)] point out that recently using in order of IL-2 and α-IFN is an effective immunization therapy mode in treatment mouse tumor model MCA-105 sarcoma and M10g cancer.The topmost discovery of this research be cytokine use in order than cytokine the time use higher effect arranged.
The adoptive immunity therapy is as the human various diseases of a kind of treatment especially treatment pattern of tumor (cancer), and its feasibility and effect are at the United States Patent (USP) of Rosenberg; The patent No. is No.4, description is arranged in 690,915.Yet as above indicate like that, need and implement such adoptive immunity therapy but toxic method when not resembling independent use IL-2.Also need a kind of at low effector cell: under the condition of target cell ratio with regard to effectively therapeutic modality.
Summary of the invention
The present invention has satisfied these requirements, provides IL-10 to use separately or share with IL-2 again and/or share to increase the particularly method of LAK and NK cell lytic activity of PBMCs with α-IFN.
More particularly, the invention provides one and use the effective dose of the IL-10 that activates PBMCs and the method that cancer is disappeared to the cancer patient.The preferred embodiment of activated PBMCs is while or follow-up use IL-10.
Be that IL-10 and IL-2 share in one embodiment, the amount of IL-2 is enough to increase the LAK cell activity, but can not cause toxic and side effects again when using separately.
Another embodiment is that α-IFN that IL-10 and being enough to increases LAK cytoactive amount share.
To also have a kind of embodiment be IL-10 and (a) be enough to increase the LAK cell activity but the IL-2 that do not cause toxic and side effects dosage when using separately again uses together and α-IFN of (b) being enough to increase LAK cytoactive amount uses together.
The present invention also further provides the method for blocking the cytotoxicity that is caused by IL-2 with endogenous interleukin-4 (IL-4) antagonism, comprises the IL-10 that uses effective dose to this class patient.
The present invention further provides and comprised acceptable carrier on medicament composition that IL-10 and IL-2 and/or α-IFN share and the pharmaceutics.
Preferred end user's IL-10, IL-2 and α-IFN in aforesaid method and composition, preferred people IL-10, the IL-2 that is to use reorganization, and α-IFN.
Detailed Description Of The Invention
All lists of references cited herein all are incorporated into as reference through overall consideration.
The present invention is one and improve uses the method for prior art that IL-2 induces the cell lysis activity of NK and LAK cell.The present invention is by removing IL-2 fully or reducing the dosage of the IL-2 that institute must use greatly and alleviated the typical toxic and side effects that is produced greatly in the method for use IL-2.
Unless special definition, used here various terms instruct the known in the art term of the present invention to have identical implication with being used to.
Resembling used term " adoptive immunity therapy " implication here promptly is to transplant the activated therapy that the immunocyte of function is arranged to patient.Preferred embodiment is to comprise the LAK and the NK cell that derive from the patient who is receiving treatment in these cells.
The implication that used here term " disappears " is the same with the common mensuration of this area, refers to measurable the reducing on one or more gross tumor volumes.
Used here " Interleukin-10 " or " IL-10 " definition are that this albumen of a kind of albumen (a) is the patent U.S.Patent Application Serial No.07/917 of the albumen (for example lacking excretory targeting sequencing) of the sophisticated IL-10 of a kind of aminoacid sequence in submission on July 20th, 1992, disclosed like that on 806, and this part patent is identical with International Application No.WO91/00349; (b) have the biologic activity identical with natural IL-10.For purpose of the present invention, (for example chemosynthesis or produce in escherichia coli) all is equivalent no matter that glycosylated IL-10 (for example for example producing in the Chinese hamster ovary celI at eukaryotic cell) does not still have is glycosylated, and can mutual alternative use.But also comprise mutein and other analog, and comprising BCRF1 albumen (the viral IL-10 of African lymphocyte virus), this albumen also has the biologic activity of IL-10.
The IL-10 that is applicable to purposes of the present invention has different sources.For example, can from this protein culture medium of activated secretion, separate acquisition.In addition, IL-10 or its active fragment can carry out chemosynthesis by the standard technique of knowing in this area.Ask for an interview Mettifield, Science 233:341 (1986) and Atherton et al., (Solid Phase) A Practical Approach, 1989, I.R.L.Press, Oxford.
The preferred version that obtains this albumen or polypeptide is to obtain by recombinant technique with the nucleic acid that separates the coding IL-10 polypeptide that obtains.The book of introducing general molecular biology method has for example Sambrook et al., (Molecular Cloning) (A Laboratory Maunal), Cold Spring Harbor, New York, 2d ed., 1989, with By Ausbel et al., (eds) (Current Protocolsin Molecular Biology), Green/Woley, New York (1987 and periodicsupplements).The technology of correct sequence available standards obtains from genomic library or cDNA library.Also can use polymerase chain reaction (PCR) to obtain.Referring to, for example (PCR Protocols): (AGuide to Methods and Application), 1990, Innis et al., (Ed.) Academic Press.New York.
The library is to use the nucleic acid construct that obtains from suitable cell, asks for an interview the manufacture method that for example promptly discloses recombinant IL-10 in InternationalAppliction Publiction No.WO91/00349.Useful gene order can be found, GenBank.andBMPL or ncleic acid and PIR and Swiss-Prot for protein for example in the data base of various sequences for example, c/oIntelligenetics, Mountain View, California, or the Genetics Computer Group, University of Wisconsin Biotechnology Center, Madison, the Wisconsin foregoing is incorporated herein as reference.
The clone who includes the sequence of coding people IL-10 has been collected in American Type CultureCollection (ATCC), Rockville, and Martyland, registration number is No. 68192,68191 and.Have coding IL-10 sequence other clones the available nucleic acid hybridization of evaluation or have for the coded albumen of expression vector and can detect with the method for immunity.According to the resulting oligonucleotide probe of being preserved among the disclosed Intemational Application Publication No.WO91/00349 of sequence is useful especially, and sequence oligonucleotide probe also can prepare from the conserved region of other species related gene.Another is selected, based on the IL-10 aminoacid sequence and the degeneration probe also can use.
The method of available standards transforms the cell line of protokaryon, mammal, yeast or insecticide, makes them express this a large amount of peptide species.The escherichia coli representative strain of expressing and the clone is suitable for is comprised: W3110 (ATCCBi, 27325), X1776 (ATCC No.31244), X2282 RR1 (ATCCMp/31343), the mammalian cell strain of representative comprises: COS-7 cell strain, mouse Lcell and CHP cell.See?Sambrook(1989)and?Ausubel?et?al.，1987?Supplement)。
Available various expression vector is expressed the DNA of coding IL-10.The common carrier of express recombinant protein all can be used in protokaryon or eukaryotic cell.Preferably carrier comprises, by Okayama et al., and Mol Cell.Biol.3:280 (1983); And Takebe et al., each carrier of the described PCD of Mol.Cell.Biol.8:466 (1988).Other carrier based on SV40 comprises that those exist: Kaufmen et al., Mol.Cell.Biol.2:1304 (1982) and U.S.Patent No.4, in 675,285 disclosed carrier these for example mouse Lcell is particularly useful to monkey COS-7 cell (ATCCNo.CRL 1651) and other mammalian cell based on the carrier of SV40.Also please see Douwelset al., (1989 and Supplements) (Cloning Vectors): (A laboratoryManual) Elsevier, New York.
IL-10 can transform or the yeast of transfection or mammalian cell in soluble form for example with the form production of secretions product.Polypeptide can be by the standard method purification of knowing in this area.For example purification step comprises: ammonium sulfate precipitation, ion-exchange chromatography, gel filtration, electrophoresis, affinity chromatograph etc.Please see Methods inEnzymology Purificaition Principles and Practiles (Springet-Vertag, NewYork, 1982).
The another kind of selection be, IL-10 also can insoluble for example polymer or the form production of inclusion body.The IL-10 of this form can carry out purification with the standard step of knowing in this area.In the example of purification step there being purification step: by centrifugal the inclusion body and the host cell of disintegrate is separated, dissolved inclusion body with chaotropic agent and Reducing agent then and make polypeptide be formed with bioactive configuration.The details of these steps is asked for an interview: Winklet et al. for example, Biochemistry, 25:4041 (1986); Winklet et al., Bio/Technology 3:9923 (1985); Kiths et al., and U.S .Patent No.4,569,790.
The nucleotide sequence that is used for transfection host cell can be modified so that IL-10 or its fragment have various desired characteristics by standard technique.This class modification makes the sequence of IL-10 and natural generation little of for example using amino acid whose insertion on the primary structure level, replaces, lacks and merge.These modification modes can be used in combination and obtain final modified protein chain.
The mutant of aminoacid sequence can comprise the increase serum half-life by different targets, makes things convenient for purification and preparation, promotes the effect of treatment, alleviates the generation of the toxic and side effects in treatment and the order of severity and is prepared.The mutant of amino acid sequence all is the non-existent but predetermined mutant of occurring in nature, though some other mutant can be the translation back for example glycosylated mutant of mutant or with bonded albumen of Polyethylene Glycol (PEQ) or the like.These mutants just can use in the present invention as long as it also remains with the biologic activity of IL-10.
To the available various technology of modification of the sequence of coded polypeptide for example the sudden change [Gillman etc., Gene 8:81 (1987)] of site guiding easily finish.Most bodies of modifying all will be estimated by the screening of routine desired feature in a suitable analysis system.For example in International ApplicationPublication No.WO91/00349, promptly introduced the active external detection method of several evaluation IL-10.
Preferably, the IL-10 that the treatment human diseases should be chosen is though the IL-10 of virus or the IL-10 that comes from other mammalss also might be used.Preferred scheme is to use the people IL-10 of recombinant.The preparation of people and mice IL-10 is existing the introduction in International Application WO91/00349.The clone of the IL-10 of epstein-barr virus (EB) (BCRF1 albumen) and expression are gone up open by Moore etal. at Science 248:1230 (1990).Recombinant people IL-10 has become a kind of commodity and can buy, for example from Prepro.Tech, and Inc., Rocky Hill, NJ buys.
Each individuality that is suitable for Therapeutic Method of the present invention comprises that any tumour patient can both benefit during particularly LAK and NK cytoactive activate from activating on the PBMC cell lysis activity.Representational cancer patient is existing to be introduced, Rosenberg for example, and the article of Supra on patent and (Scientific American) is described.Therapeutic scheme of the present invention also is applicable to anticipates so that the individuality that endogenous IL-4 level raises, like this so that IL-4 has blocked the activation of IL-2 pair cell lytic activity.Preferred therapeutic scheme is to use earlier the IL-10 pretreatment before using IL-2 in this class individuality.
Standard method and the technology in order to preparation IL-10 introduced among the present invention also can be used for preparing IL-2 and α-IFN, the IL-2 that uses among the present invention and α-IFN can obtain also that (for example IL-2 can be from Cetus from commodity approach, Corporation, Emeryville, CA, α-IFN can be from Schenng, Corp., Keniluorth, NJ obtains).
Except the IL-2 of IL-10 and attenuating level share and/or α-IFN Clicks here the method introduced uses together again, the external activation of PBMCs (preferred version is to obtain with standard method from the patient that will receive treatment) and the use of these cells are undertaken by the document of Rosenberg above-mentioned basically.The quantitative range of employed activated PBMCs is 10
6～10
12Individual.Although optional, embodiment preferred is to use or follow-up use with activated like this cell as IL-10 described here.
Be in one embodiment earlier with IL-10 preconditioning stimulated PBMCs[for example 4ng/ml (100 units/ml) or 40ng/ml people IL-10 in the presence of cultivated about 3 days at 37 ℃], then, washed cell is removed the IL-2 (being typically about 2units/m) that free IL-10 adds low concentration again.
Here used by 1L-10 separately or to use the preferred operational version with the active cell dissolved cell with other cytokines be to use intravenous injection.This also can be carried out with the following method, for example enters big peripheral vein or enters Hepatic artery by the conduit of percutaneous by central venous catheter.
IL-10 uses with the form of medicament composition usually, and said composition comprises that the IL-10 or the IL-10 of the effective dose of medicament carrier and single usefulness share with IL-2 and/or α-IFN.Medicament carrier can be any compatible non-toxicant that is suitable for medicament is transferred to the patient, the existing very dark understanding of useful composition to this class medicine without digestive tract use approach, example is referring to Remingtons PhermaceuticalScience 15th Ed. (Mack Publishing Company Easton, PA, 1980).In addition, compositions of the present invention can enter in patient's body by drug delivery system that implant or injection.For example, Urquhert et al., Ann.Rev.Pharmacol.Toxicol.24:99 (1984); Leuis (Ed.) (Controlled Release of Pesticides and Pharmeceuticals) (PlenumPress, NJ.1981) U.S.Patent No.3,270,960; Or the like.
The use of cytokine can comprise vein, peritoneum and subcutaneous administration by any use approach of knowing.Intravenously administrable is most preferred use approach.
When by the administration of non-digestive tract mode, compositions is all made the injectable forms (solution, suspension, emulsion) of unified unit dose in conjunction with medicament carrier.The example of this class carrier has normal saline, Ringer ' s liquid, Glucose Liquid and Hank ' s liquid.The carrier of nonaqueous phase can be used fixed oil and ethyl oleate.Preferred vector is 5% dextrose.Carrier can comprise some a spot of additives to increase isotonicity and chemical stability for example buffer agent and antiseptic etc.The preferred Unified Form of IL-10 is to be substantially free of polymer and other proteic pure form, and concentration is about 5-20 μ g/ml.
The compositions of the present invention also gene therapy technology of available standards is transmitted and is imported in patient's body, comprise as direct DNA tissue injection, use the implantation of recombinant viral vector and transfectional cell, for example see, Rosenberg, J.Clin.Oncol.10:180 (1992).
The share of one or more healing potions that this place is introduced can be used (using with IL-10) simultaneously or can be used in order.Preferred version is that IL-10 uses prior to IL-2.α-IFN can be with IL-10 and/or IL-2 or follow-up use.All medicaments have enough concentration so that effectively make tumor regression in treatment in patient's body.
The implication of used here " effective dose " term refers to that the amount of IL-10 is enough to reduce or stops side effect in the adoptive immunity therapy, can improve the cell lysis activity of LAK and NK cell simultaneously again.The effective dose of required cytokine can be according to resembling the tumor type and the state of being treated concerning a special patient, a class factor such as the amount of the order of severity of patient's holistic health, used Therapeutic Method, side effect and the other medicines that using simultaneously and kind and different.
Amount the definition here of the cytokine that " is enough to increase the LAK cell activity " refers to that with the Daudi cell be the cytolysis appraisement system, the cell lysis activity that cell concentration produced that needs can increase by 25% during at least than the independent use of IL-10, preferred version is to be increased at least 50%, and most preferably scheme is to be increased at least about 100%.
Preferred version is the threshold dose of its maximum of IL-10 administrable, from 10～10
8Unit per kilogram of body weight every day.IL-2 and α-IFN administration similarly also can use the maximum limit consumption (for example to IL-2 intravenously administrable institute consumption to be: per 8 hours 10
5The unit per kilogram of body weight uses 50ml 0.4% normal saline and 5% albumin as carrier; The dosage of α-IFN is: per 8 hours 10 of intravenously administrable
6The U per kilogram of body weight adds 5% albumin as carrier with 0.9% normal saline).The front said that used dosage was to be done to descend by the maximum limit that the clinicist can bear according to each individual patients to adjust.
Method of the present invention also can be used with traditional treatment method for cancer, and for example radiation and chemotherapy uses the chemotherapy of traditional chemotherapeutic image vinblastine, platinum-like compounds and 5-fluorouracil.
Use IL-10 separately or share the cell lysis activity enhancing of handler's peripheral blood lymphocytes initiation people's tumor target cell with other cytokine.Because the efficient of immunization therapy depends on the burden degree of tumor to a great extent, so after the main tumor mass of bulk is cut, use the Therapeutic Method described here will be useful especially.Usually the inflammatory reaction that takes place at surgical site perhaps can be beneficial to therapeutic effect.
Embodiment
Following nonlimiting examples will be used for explanation of the present invention.
IL-10 is to the influence of human peripheral blood mononuclear cell's cell lysis activity
Studied the influence of IL-10 to human peripheral blood mononuclear cell's cell in vitro lytic activity.The result is summarized as follows:
The activity of activated killer cell of lymphokine (LAK) and natural killer cell (NK) in the I.IL-10 stimulation human peripheral blood mononuclear cell.Rat anti IL-10 monoclonal antibody can in and the cell lysis activity that drives of IL-10.
2. the IL-10 that derives from CHO and escherichia expression system shows identical concentration-response pattern in LA stimulation K and NK cytoactive, be equivalent on biological significance therefore.
3.PBMCs the activity when handling the LAK cytoactive that is shown and make separately than any cytokine with IL-10 and low dosage IL-2 is all strong.
4.PBMCs add the lytic activity of IL-2 cell after 2 days again increases with the IL-10 pretreatment.
5.IL-4 suppress the inductive LAK cytoactive of IL-2, and this inhibitory action of IL-10 antagonism.
6.IL-10 add that IL-2 is at low effector cell: the lytic activity that produces the LAK cell pair cell that increases under the situation of target cell ratio.
Except having under the condition of IL-10 endothelial cells cultured unaffected to replying of exogenous factor (being γ-IFN and TFN-α) to also finding the viewed influence of PBMCs, and in containing the culture medium of IL-2 endothelial cells cultured because the toxicity of IL-2 and Fails To Respond.
Materials and methods
The human cell factor of reorganization and the antibody of anti-IL-10
The people IL-10 of reorganization with standard method produce (from escherichia coli and CHO the two).The specific activity that detects with product behind the standard method purification with the propagation of MC-9 cell is respectively 4.1 * 10
7E.coli and 2.1 * 10
7(CHO) Units/mg[Thompson-Sripes et al., J.Exp.Med.173:507 (1991)], in this way by being defined as the biologic activity that the single basically IL-10 of about 4ng contains 100 units approximately.
Obtain a monoclonal antibody of naming to the rat anti people IL-10 of 19F1 from Palo Alto.CA DNAX molecular biology research John doctor Abrams of institute.
Human peripheral blood mononuclear cell's separation (PBMCs)
Peripheral blood is to obtain with adult's donor of venipuncture from health as anticoagulant with heparin or EDTA.PBMCs is isolating with two step method, comprise glucosan precipitation and according at FICOLL PAQUE " go up with 1250rpm (rev/min) centrifugal 30 minutes.Collection mainly comprises lymphocyte and monocytic interface zone, and cleans secondary (JRHBiosciences) at least with the RPMI culture medium (complete medium) that contains 10% hyclone.
Cytotoxicity detects
Daudi (to the LAK sensitivity) and K562 (to the NK sensitivity) target cell obtain with registration number (underaccession Nos.) CCL 213 and CCL 243 respectively from American type culture collection (American Type Tissue Collection) respectively.With Spits etc. [J.Immunol141:29 (the 1989)] method of introducing Daudi and K562 cell marking with
51Cr.PBMCs collects through after cultivating, and washes secondary, can be at one
51Action effect daughter cell in the Cr release detection architecture (Spits etal., Supra).With 5 * 10
3 51The target cell of Cr labelling and not commensurability effector cell (E/T=20: 1,5: 1,2: 1) in 96 orifice plates at the bottom of the V-type, mix with 100 μ l volumes.96 orifice plates are with 1000rpm CO in saturated humidity 5% after centrifugal 5 minutes
2Cultivated 4 hours in the gas, cultivate and use centrifugal 5 minutes of 500xg after 4 hours.Supernatant is collected with SKATRON (Skatron Instruments) catcher, and counts with gamma counter (LKB-Pharmacia).With
51The target cell of Cr labelling and 1%SDS measure total lysis by hatching.The meansigma methods of data represented three mensuration.
The percentage ratio of lysis is calculated as follows:
Human PBMC s and cytokine are hatched
A.IL-10 is hatched separately
Unless otherwise specified, the above-mentioned isolating PBMCs of institute is with every milliliter 1 * 10
6The concentration of cell contains in the 10% hyclone RPMI-1640 culture medium 37 ℃ at the IL-10 that adds IL-10 or people (CHO) cultivated 3 days.Determine cell lysis activity by the above.
B. hatch simultaneously with IL-10 and IL-2
PBMCs with the IL-10 of 4ng/ml with or do not hatch 3 days at 37 ℃ with IL-2 (Genzyme) (2 or 20U/ml).
C. cultivate with the order of IL-10 and IL-2
PBMCs was hatched in complete medium 2 days with the IL-10 of 4ng/ml.It is 2U/ml or 20U/ml that adding IL-2 makes ultimate density.After the overnight incubation, measure the cell lysis activity of LAK and NK cell.
The monoclonal antibody of anti-IL-10 activates the influence of LAK and NK cell to IL-10
Anti-IL-10 monoclonal antibody (19F1) or homotype at 2 μ g/ml contrast under the condition of (rat IgG2a) existence, and PBMCs and 40ng/ml IL-10 were hatched 3 days.
IL-10 is to the influence of activated killer cell of lymphokine among the human peripheral blood mononuclear cell and natural killer cell cell lysis activity
According to used tumor target cell from operating the cell lysis activity that can distinguish LAK and NK cell.Deriving from the lymphadenomatous Daudi cell of Burkitt ' s is the target cell of the activated LAK cell of the detection lytic activity of classics.The cell that derives from human erythroleukemia K562 cell strain is used as the specific target cell that detects NK cell lytic activity.The standard of in these experiments, people's PBMCs being introduced above the reuse with the IL-10 of variable concentrations processing in 3 days
51The release algoscopy of Cr is determined cell lysis activity.
The active result of LAK is as shown in table 1, and wherein standard deviation provides below meansigma methods.
(% with lysis represents table 1:IL-10 to the excitation of the activated PBMCs of lymphokine
a)
IL-10 concentration (ng/ml)
b
Donor 0 0.04 0.4 4 40 100
1 0 4.25 18.3 44 26.2 67.5
2 0 5.5 7.2 10 31.5 27.6
3 18 17.7 29.2 39.1 29.7 55.1
4 0 8.8 10.2 22.2 35.8 35.9
5 4.6 8.1 15.6 20.6 20.1 12.1
6 14.6 19.7 40.7 54.4 42.9 43.4
Meansigma methods 6.2 10.6 20.2
c31.7
c31.0
c40.2
c
±3.3?±2.6 ±5.1 ±6.8 ±3.2 ±8.0
A.
51The lysis percentage ratio of the Daudi target cell of Cr labelling be 20: 1 effector cellular targets cell than and obtain with the chromium release assay method of standard.The meansigma methods of data represented three mensuration.
B. before measuring cell lysis activity, the human PBMC s that obtains from normal donor was handled 3 days with IL-10 earlier.
C. use Student ' s-t check to determine between IL-10 processed group and medium controls significant difference is arranged, P≤0.05.
The data of table 1 shows that IL-10 is to the active influence of LAK among the PBMCs that obtains from 6 people's donors.Although the concentration that significant difference between 6 donors, the increase of the cytolysis ability that IL-10 brings out in all 6 donors all show IL-10 relies on.The concentration of the active significant IL-10 of statistics is 0.4ng/ml or bigger (P≤0.05).But also observe the basic level (promptly in acellular factor condition) that donor PBMCs has shown a cell lysis activity, effector cell that all were measured and target cell than in 5% or still less cell lysis activity to IL-10 reaction the sensitiveest (data is not enclosed)
To other tumor target cell, comprise the strain of human kidney cells tumor, two different Humanmachine tumour strains and human colon cancer cell strain also obtain similar result.Rosenberg also once used remarkable renal cell carcinoma and melanoma cell, but also the patient who has such tumor was done intravital adoptive immunotherapy with IL-2.In all experiments, the molten cytosis percentage ratio of target cell all relies on IL-10 concentration.
In other some experiments, find IL-10 single with or share with IL-2 and also can cause increase U937 (human tissue cell's tumor); The cell lysis activity of SW620 (human colon carcinoma) and SKBR3 cell (human breast carcinoma).IL-10 does not show initiation reaction in the dosage of being tested in the experiment of HS294T cell (Humanmachine tumour) when doing target cell.
The cell lysis activity of the NK of foundation level (promptly has the high trend of molten cytoactive than LAK cell base level to the lysis of K562 target cell under the condition of factor-containing not.Active available cytokine such as the IL-2 of NK further strengthen [Perussia, Supra; Philips and Lanier J.Exp.Med164:814 (1986)].
In the parallel laboratory test of measuring the LAK cytoactive, measured the influence of IL-10 to NK cytoactive among the PBMCs with above-mentioned 6 identical donors.The result is as shown in table 2, and wherein standard deviation provides below meansigma methods.
Table 2:IL-10 excites to NK among the PBMCs is active that (% with lysis represents
a)
The concentration of IL-10 (ng/ml)
b
Donor 0 0.04 0.4 4 40 100
1 22.2 30.6 25.2 57.2 51.5 49.7
2 20.4 24.7 31.3 56.7 65.6 71.5
3 61.4 55.6 37.9 81.1 80.0 81.5
4 13.8 25.2 23.2 37.5 52.2 54.2
5 13.0 19.9 20.2 25.1 23.5 20.3
6 15.9 25.3 45.4 66.6 43.7 56.1
Meansigma methods 24.4 30.2 30.5 54.0
c52.75
c55.5
c
±7.5 ±5.2 ±3.9 ±8.2 ±7.8 ±8.5
A.
51The lysis percentage ratio of the Daudi target cell of Cr labelling is to be that the chromium release assay method with standard obtains under 20: 1 the condition at effector cell and target cell ratio.Data are meansigma methodss of three experiments.
B. the human PBMC s from normal donor source handled 3 days with IL-10 earlier before measuring cell lysis activity.
C. use Student ' s-t check to determine significant difference is arranged, P≤0.05 between IL-10 processed group and medium controls.
Can see obviously that from table 2 IL-10 induces the enhancing of the cell-mediated cytotoxicity of from the PBMCs of donor source NK that significant dose dependent is arranged.The concentration of IL-10 is that 4ng/ml or the lysis when higher obviously increase statistically.As in the LAK determination of activity, IL-10 is different and variant according to donor to the influence of NK cytoactive.
IL-2 and IL-10 are to the comparison of the influence of endotheliocyte
Designed the experiment of in the presence of IL-10, measuring the survival ability of endotheliocyte monolayer.When endotheliocyte is cultivated under the IL-10 existence condition replying of the foreign cell factor (γ-IFN and TFN-α) do not weakened, and parallel laboratory test with the IL-2 of same unit dosage with cultivating the identical time because the toxicity of IL-2 makes cell lose the responsibility of the foreign cell factor.
The monoclonal antibody of anti-IL-10 activates the influence of LAK and NK cytosis to IL-10
The meansigma methods of expression has standard deviation shown in table 3 and table 4, in the presence of the monoclonal antibody 19F1 of 2 μ g/ml IL-10, uses activation minimizing 3 times (P≤0.05) of the IL-10 of 40ng/ml to the cell lysis activity of LAK and NK cell respectively.Replace with the rat IgG2a homotype control antibodies of 2 μ g/ml anti-IL-10 monoclonal antibody the result of the horizontal gained of celliferous lytic activity and those can not draw difference statistically with 40ng/ml IL-10 gained result separately.
Table 3: the monoclonal antibody of anti-IL-10 is to the inhibition (% represents with lysis) of the cell lysis activity of the activated killer cell of the inductive lymphokine of IL-10
a
Condition of culture
b
Donor culture medium 40ng/ml IL-10 IL-10
IL-10 +19F1 +IgG2a
1 5.6 23.5 15.7 28.2
2 4.7 21.3 11.0 17.5
3 5.4 42.5 0.0 35.8
4 2.1 42.0 12.8 26.5
5 4.7 19.1 0.0 11.65
Meansigma methods 4.5 ± 0.6 29.6 ± 5.3 7.9c ± 3.3 23.9 ± 4.3
A.
51The lysis percentage ratio of the Daudi target cell of Cr labelling is at 20: 1 effector cells: target cell than and with the chromium method for releasing of standard mensuration.Data are three meansigma methodss of measuring.
B. the human peripheral blood mononuclear cell who obtains from normal donor handled 3 days with 40ng/ml IL-10 under the condition of 2mg/ml19F1 (anti-people IL-10) or rat IgG2a (homotype contrast) existence earlier before surveying cell lysis activity.
C. use Student ' s-t check IL-10 individual processing and anti-IL-10 monoclonal antibody exist handle between the result significant difference is arranged, P≤0.05.
Table 4: (% represents anti-IL-10 monoclonal antibody with lysis to the neutralization of the inductive natural killer cell activity of IL-10
a)
Condition of culture
b
Donor culture medium 40ng/ml IL-10 IL-10
IL-10 +19F1 +IgG2a
1 21.1 38.3 20.6 27.2
2 28.0 71.0 22.2 53.2
3 22.2 51.5 0.0 70.5
4 18.0 25.4 12.3 20.8
5 23.2 53.2 54.5 84.9
Meansigma methods 22.5 ± 1.6 47.9 ± 7.6 19.1c ± 6.6 51.3 ± 12.7
A.[
51Cr] the Daudi target cell lysis percentage ratio of labelling is at 20: 1 effector cells: target cell than and with the chromium method for releasing mensuration of standard.Data are three meansigma methodss of measuring.
B. the human peripheral blood mononuclear cell who obtains from normal donor handled 3 days with 40ng/ml IL-10 under the condition of 2mg/ml19F1 (anti-people IL-10) or rat IgG2a (homotype contrast) existence earlier before surveying cell lysis activity.
C. use Student ' s-t check IL-10 individual processing and anti-IL-10 monoclonal antibody exist handle between the result significant difference is arranged, P≤0.05.
IL-10 and other cytokine are share inductive cell lysis activity
A.IL-10 and IL-2 are hatched simultaneously
Human PBMC s was at 5: 1 effector cell: target cell than and IL-10 down the IL-2 of usefulness time limit activation concentration (2 or 20U/ml) hatch, the result is as shown in table 5, is standard deviation under the meansigma methods.
Table 5: (% represents with lysis to hatch the activated cell lysis activity of the inductive lymphokine of institute simultaneously with IL-10 and IL-2 in the human peripheral blood mononuclear cell
a)
Condition of culture
b
Donor culture medium 2U IL-2 4ng IL-10 IL-10 20U IL-2
+IL-2
1 0.0 3.3 3.7 15.9 24.6
2 2.6 7.6 3.8 23.5 41.0
3 6.1 5.0 13.3 17.7 11.4
4 3.3 50.1 51.6 68.7 80.0
5 2.4 9.4 12.8 29.3 45.7
6 9.9 28.9 16.0 38.8 67.1
Meansigma methods 4.1 17.4 16.8 32.3 44.9c
±1.42 ±7.6 ±7.2 ±7.2 ±10.4
A.[
51Cr] the lysis percentage ratio of Daudi target cell of labelling is the effector cell at 5: 1: target cell than and with the chromium method for releasing mensuration of standard.Data are three meansigma methodss of measuring.
B. the human peripheral blood mononuclear cell from normal donor source was handled 3 days with 4ng/mlIL-10 and 2U/ml IL-2 earlier before surveying cell lysis activity.
C. using Student ' s-t check to measure donor PBMCs handles with IL-10 and IL-2 and handles relatively there was no significant difference between the cell lysis activity, P≤0.05 with 20U/mlIL-2 is single.
As shown in table 5,2U/ml and 4ng/ml IL-10 share and make LAK activity (effector cell: target cell=5: 1) extra increase is arranged.This activity level all significantly increases than the activity of single time spent of any cytokine in two kinds.Reached the activity level that 10 times high IL-2 concentration is produced.Using Studeat ' s-t check to record the result is significant statistically, P≤0.05.
B.IL-10 and α-IFN are hatched simultaneously
The cell lysis activity of hatching jointly the Daudi cell with PBMCs IL-10 and α-IFN shows as similar extra increase, but to the NK target cell is not.The result is as shown in table 6, and standard deviation provides below meansigma methods.
Table 6: in the human peripheral blood mononuclear cell, share the activated killer cell activity of lymphokine induced that (% represents with lysis with IL-10 and α-IFN
a)
Condition of culture
b
Donor culture medium α-IFN IL-10 IL-10+ α-IFN
1 4.4 7.9 17.8 35.7
2 1.0 11.7 15.4 32.0
3 1.6 12.3 5.5 26.4
Meansigma methods 2.3 10.6 12.6 31.4
±1.0 ±1.4 ±3.8 ±2.7
A.[
51Cr] the lysis percentage ratio of Daudi target cell of labelling is the effector cell at 10: 1: target cell than and with the chromium method for releasing mensuration of standard.Data are three meansigma methodss of measuring.
B. the human peripheral blood mononuclear cell from normal donor preparation handled 3 days with 4ng/mlIL-10 and 100U/ml α-IFN earlier before measuring cell lysis activity.
C. use Student ' s-t check to predict, IL-10 and α-IFN share with IL-10 or the independent use of α-IFN relatively induce on the cell lysis activity of donor PBMCs viewed different be significant statistically, P＜0.05.
On the contrary, IL-10 uses with IL-4, IL-5, GMCSF or γ-IFN and uses more effective (result is unlisted) separately unlike IL-10.
The order of IL-10 and IL-2 is hatched
By the method for introducing above, PBMCs is remained on culture medium or be added with in the culture medium of IL-10.Add IL-2 two days later, making its final concentration is 2 or 20U/ml.After incubated overnight again, measure cytotoxic activity to the Daudi cell.As shown in table 7 from the result that 5 donors are comprehensive, wherein standard deviation provides below meansigma methods.
Table 7: in the human peripheral blood mononuclear cell, then handle the activated cell lysis activity of lymphokine induced that (% represents with lysis with IL-2 with the IL-10 pretreatment
a)
Pre-condition of culture
b
Donor culture medium IL-10
cIL-2
dIL-10+IL-2
e
1 29.7 21.1 44.8 116
2 5.5 18.3 12.2 20.7
3 14.7 58.1 74.5 100
4 26.2 59.5 54.3 81.9
5 4.8 28.2 22.5 40.6
Meansigma methods 16.2 37.0 41.7 71.8
±5.1 ±9.0 ±11.4 ±17.9
A.[
51Cr] percentage ratio of lysis of Daudi target cell of labelling is the effector cell at 5: 1: target cell than and with the chromium method for releasing mensuration of standard.Data are three meansigma methodss of measuring.
B. the human peripheral blood mononuclear cell from normal donor preparation kept 2 days the culture medium that is being added with earlier 4ng/ml IL-10 before the adding 2U/ml IL-2.Measuring cell lysis activity after the incubated overnight again.
C. donor PBMCs was hatched 3 days with IL-10.
D. donor PBMCs is that single culture is in culture medium before adding IL-2.
E. donor PBMCs was hatched 2 days with IL-10 earlier before adding 20U IL-2.
As shown in table 7, the viewed cell lysis activity of pretreated experimental group (71.8 ± 17.9%) has increased about 2 times than the cell lysis activity (41.7 ± 11.4) of only donor PBMCs being cultivated the experimental group in complete medium before adding IL-2 to donor PBMCs with IL-10 earlier before adding IL-2.Observed between pretreated sample with IL-10 pretreatment and those without IL-10 be significant P≤0.14 statistically to difference.
In then hatching in proper order, see the pattern (data is not shown) that similar cell lysis activity increases with α-IFN with IL-10.
IL-10 and IL-4 are to the blocking-up of the inductive cytotoxicity of IL-2
The PBMCs that obtains from 6 human donors is cultivated culture medium, and culture medium contains respectively: the IL-2 of 20U/ml is singly arranged, and the IL-2 of 20U/ml adds the IL-4 of 1000U/ml, and 20U/ml IL-2 adds 1000U/ml IL-4 and adds 4ng/ml IL-10.The result is as shown in table 8, and standard deviation provides below meansigma methods.8:IL-10 is activated to the inductive lymphokine of IL-2 to IL-4 among the human peripheral blood mononuclear cell for table
(% represents the antagonism of the blocking-up of cell lysis activity with lysis
a)
Donor culture medium IL-2
bIL-2
c+ IL-2
d+
IL-4 IL-4+
IL-10
1 12.7 27.6 35.0 45.7
2 5.4 26.3 17.5 33.7
3 1.7 25.1 14.1 36.0
4 3.8 29.6 14.1 22.1
5 3.3 47.1 17.1 63.8
6 6.6 49.5 20.6 40.8
Meansigma methods 5.5 34.2 19.7 40.3
±1.6 ±4.5 ±3.2 ±5.7
A. to [
51Cr] the Daudi target cell lysis percentage ratio of labelling is the effector cell at 20: 1: target cell than and measure with the chromium method for releasing of standard.Data are three meansigma methodss of measuring.
B. the human peripheral blood mononuclear cell who is separated to from normal donor handled 3 days with 20U/ml IL-2.
C. donor PBMCs handled 3 days with 20U/ml IL-2 and 100U/ml people IL-4.
D. donor PBMCs 20U/ml IL-2,1000U/ml people IL-4 and 4ng/ml IL-10 processing.
Show independent with 20U/ml IL-2 processing, the lysis ratio about 34.2% of the target cell of LAK sensitivity as the data in the table 8.When adding the IL-4 of 1000U/ml when hatching beginning, the lysis ability has been suppressed about 2 times.If but added the IL-10 of 4ng/ml in initial 24 hours of cultivating, the inhibitory action of IL-4 would not just have.
The there was no significant difference as a result that the IL-2 list is used and all three kinds of cytokines are used together (Student ' the s-t check, P≤0.05), illustrate that IL-10 is basically completely to the antagonism of IL-4 blocking effect.
As long as without prejudice to spirit of the present invention and scope, the present invention can do various modifications and changes, and this point is apparent to one skilled in the art.The specific embodiments of being introduced just provides with the form of giving an example here, and the present invention is limited by the appended claim clause in back also just.
Claims (12)
- One kind the treatment method for cancer, comprise to the cancer patient use effective dose the activated PBMCs of IL-10 so that this cancer disappear.
- 2. the method for claim 1 further comprises the IL-10 that simultaneously or sequentially said individuality is used effective dose.
- 3. the production method of the medicament composition of treatment cancer comprises that activated PBMCs of IL-10 and pharmacology are gone up acceptable carrier to be mixed.
- 4. arbitrary method of claim 1 to 3, wherein IL-10 use be with (a) q.s can strengthen the LAK cytoactive but do not cause the IL-2 of toxic and side effects and/or (b) the active α-IFN of LAK cell-stimulating that can increase of q.s share.
- 5. the medicament composition of treatment cancer comprises acceptable carrier on activated PBMCs of IL-10 and the pharmacology.
- 6. the medicament composition of claim 5, further comprise IL-10 single with or share with IL-2 and/or α-IFN.
- 7.IL-10 the application of activated PBMCs aspect the treatment cancer.
- 8.IL-10 the application of activated PBMCs aspect the medicament of making the treatment cancer.
- 9. claim 7 or 8 application, wherein activated PBMCs of IL-10 and independent IL-10 use together or use with IL-10, IL-2 and/or α-IFN.
- 10. an antagonism endogenous IL-4 comprises the IL-10 that the patient that this class treatment demand is arranged is used effective dose to the method for the blocking effect of the inductive cytotoxicity of IL-2.
- 11.IL-10 making antagonism IL-4 to the application aspect the medicament of the blocking effect of the inductive cytotoxicity of IL-2.
- 12. the method for each of claim 1 to 11, pharmaceutical composition or application, IL-10 wherein is people IL-10.
Priority Applications (10)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| SK916-96A SK91696A3 (en) | 1994-01-20 | 1994-01-20 | Use of il-10 to stimulate peripheral blood mononuclear cell cytolytic activity |
| EP94904303A EP0740552A1 (en) | 1994-01-20 | 1994-01-20 | Use of il-10 to stimulate peripheral blood mononuclear cell cytolytic activity |
| CN94194863A CN1142186A (en) | 1994-01-20 | 1994-01-20 | Use of IL-10 to stimulate peripheral blood mononuclear cell cytolytic activity |
| AU58425/94A AU707019B2 (en) | 1994-01-20 | 1994-01-20 | Use of IL-10 to stimulate peripheral blood mononuclear cell cytolytic activity |
| NZ259584A NZ259584A (en) | 1994-01-20 | 1994-01-20 | Cancer treatment using interleukin 10 activated peripheral blood mononuclear cells optionally combined with il-10 alone or in combination with il-2 or alpha interferon (alpha-ifn), to stimulate peripheral blood mononuclear cells |
| PL94315513A PL175343B1 (en) | 1994-01-20 | 1994-01-20 | Application of il-10 for cytolytical activity stimulation of mononuclear peripheral blood cells |
| PCT/IB1994/000008 WO1995019780A1 (en) | 1994-01-20 | 1994-01-20 | Use of il-10 to stimulate peripheral blood mononuclear cell cytolytic activity |
| JP7519439A JPH09508116A (en) | 1994-01-20 | 1994-01-20 | Use of IL-10 to stimulate peripheral blood mononuclear cytolytic activity |
| FI962813A FI962813A (en) | 1994-01-20 | 1996-07-11 | The use of IL-10 to stimulate the cytolytic activity of peripheral blood mononuclear cells |
| NO963029A NO963029L (en) | 1994-01-20 | 1996-07-19 | Use of IL-10 to stimulate cytological activity of single-core peripheral blood cells |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN94194863A CN1142186A (en) | 1994-01-20 | 1994-01-20 | Use of IL-10 to stimulate peripheral blood mononuclear cell cytolytic activity |
| PCT/IB1994/000008 WO1995019780A1 (en) | 1994-01-20 | 1994-01-20 | Use of il-10 to stimulate peripheral blood mononuclear cell cytolytic activity |
Publications (1)
| Publication Number | Publication Date |
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| CN1142186A true CN1142186A (en) | 1997-02-05 |
Family
ID=37708148
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN94194863A Pending CN1142186A (en) | 1994-01-20 | 1994-01-20 | Use of IL-10 to stimulate peripheral blood mononuclear cell cytolytic activity |
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| Country | Link |
|---|---|
| EP (1) | EP0740552A1 (en) |
| JP (1) | JPH09508116A (en) |
| CN (1) | CN1142186A (en) |
| AU (1) | AU707019B2 (en) |
| FI (1) | FI962813A (en) |
| NO (1) | NO963029L (en) |
| NZ (1) | NZ259584A (en) |
| PL (1) | PL175343B1 (en) |
| SK (1) | SK91696A3 (en) |
| WO (1) | WO1995019780A1 (en) |
Cited By (1)
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|---|---|---|---|---|
| CN101631560B (en) * | 2006-09-28 | 2013-12-25 | 默沙东公司 | Use of pegylated IL-10 to treat cancer |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2356010A1 (en) * | 1998-12-22 | 2000-06-29 | Schering Corporation | Treatment of hepatitis c virus infections with interleukin-10 |
| ES2367891T3 (en) | 2000-09-29 | 2011-11-10 | Schering Corporation | INTERLEUCINA-10 PEGILADA. |
| RU2322452C2 (en) * | 2006-03-27 | 2008-04-20 | Михаил Николаевич Смирнов | Immunomodulating composition |
| US20110044939A1 (en) * | 2007-06-27 | 2011-02-24 | Joslin Diabetes Center, Inc. | Regulatory t cells in adipose tissue |
| EP3348281B1 (en) | 2008-12-17 | 2023-07-05 | Merck Sharp & Dohme Corp. | Mono- and di-peg il-10 production; and uses |
| JP2016519108A (en) | 2013-04-18 | 2016-06-30 | アルモ・バイオサイエンシーズ・インコーポレイテッド | Method for using interleukin-10 for the treatment of diseases and disorders |
| WO2014204816A2 (en) | 2013-06-17 | 2014-12-24 | Armo Biosciences, Inc. | Method for assessing protein identity and stability |
| US10010588B2 (en) | 2013-08-30 | 2018-07-03 | Armo Biosciences, Inc. | Methods of using pegylated interleukin-10 for treating hyperlipidemia |
| KR20160079114A (en) | 2013-11-11 | 2016-07-05 | 아르모 바이오사이언시스 인코포레이티드 | Methods of using interleukin-10 for treating diseases and disorders |
| WO2015187295A2 (en) | 2014-06-02 | 2015-12-10 | Armo Biosciences, Inc. | Methods of lowering serum cholesterol |
| JP2017536098A (en) | 2014-10-14 | 2017-12-07 | アルモ・バイオサイエンシーズ・インコーポレイテッド | Interleukin-15 composition and use thereof |
| WO2016064817A1 (en) | 2014-10-22 | 2016-04-28 | Armo Biosciences, Inc. | Methods of using interleukin-10 for treating diseases and disorders |
| US10618970B2 (en) | 2015-02-03 | 2020-04-14 | Armo Biosciences, Inc. | Method of treating cancer with IL-10 and antibodies that induce ADCC |
| AU2016268403A1 (en) | 2015-05-28 | 2017-12-07 | Armo Biosciences, Inc. | Pegylated interleukin-10 for use in treating cancer |
| JP7053453B2 (en) | 2015-08-25 | 2022-04-12 | アルモ・バイオサイエンシーズ・インコーポレイテッド | How to use interleukin 10 to treat diseases and disorders |
| WO2023072861A1 (en) * | 2021-10-25 | 2023-05-04 | Ellennbe Gmbh | Pharmaceutical composition and kit comprising an immunomodulatory substance for treating diseases |
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| MX9200161A (en) * | 1991-01-16 | 1992-07-01 | Schering Corp | USE OF INTERLEUKIN-10 IN ADOPTIVE CANCER IMMUNOTHERAPY |
| JP3260368B2 (en) * | 1991-01-16 | 2002-02-25 | シェリング・コーポレーション | Treatment of neoplastic disease with interleukin-10 |
| US5055045A (en) * | 1991-03-18 | 1991-10-08 | Dickie Robert G | Disposable dental matrix retainer clamp |
-
1994
- 1994-01-20 SK SK916-96A patent/SK91696A3/en unknown
- 1994-01-20 WO PCT/IB1994/000008 patent/WO1995019780A1/en not_active Application Discontinuation
- 1994-01-20 NZ NZ259584A patent/NZ259584A/en unknown
- 1994-01-20 JP JP7519439A patent/JPH09508116A/en active Pending
- 1994-01-20 PL PL94315513A patent/PL175343B1/en unknown
- 1994-01-20 EP EP94904303A patent/EP0740552A1/en not_active Withdrawn
- 1994-01-20 CN CN94194863A patent/CN1142186A/en active Pending
- 1994-01-20 AU AU58425/94A patent/AU707019B2/en not_active Ceased
-
1996
- 1996-07-11 FI FI962813A patent/FI962813A/en unknown
- 1996-07-19 NO NO963029A patent/NO963029L/en unknown
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101631560B (en) * | 2006-09-28 | 2013-12-25 | 默沙东公司 | Use of pegylated IL-10 to treat cancer |
Also Published As
| Publication number | Publication date |
|---|---|
| FI962813A0 (en) | 1996-07-11 |
| EP0740552A1 (en) | 1996-11-06 |
| NO963029L (en) | 1996-09-19 |
| NZ259584A (en) | 1998-01-26 |
| NO963029D0 (en) | 1996-07-19 |
| FI962813A (en) | 1996-07-11 |
| PL175343B1 (en) | 1998-12-31 |
| AU5842594A (en) | 1995-08-08 |
| WO1995019780A1 (en) | 1995-07-27 |
| AU707019B2 (en) | 1999-07-01 |
| JPH09508116A (en) | 1997-08-19 |
| PL315513A1 (en) | 1996-11-12 |
| SK91696A3 (en) | 1997-04-09 |
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