WO1989005657A1 - Activation de cellules par lymphokine pour l'immunotherapie adoptive, par exemple de l'infection par l'hiv - Google Patents

Activation de cellules par lymphokine pour l'immunotherapie adoptive, par exemple de l'infection par l'hiv Download PDF

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Publication number
WO1989005657A1
WO1989005657A1 PCT/GB1988/001134 GB8801134W WO8905657A1 WO 1989005657 A1 WO1989005657 A1 WO 1989005657A1 GB 8801134 W GB8801134 W GB 8801134W WO 8905657 A1 WO8905657 A1 WO 8905657A1
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Prior art keywords
lymphocytes
cells
patient
activated
infectious agent
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PCT/GB1988/001134
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English (en)
Inventor
James C. Sharp
Abdul Jabear Sultan
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Browning's Clinical Pathology Services Limited
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Priority claimed from GB878729410A external-priority patent/GB8729410D0/en
Priority claimed from GB888817082A external-priority patent/GB8817082D0/en
Priority claimed from GB888823897A external-priority patent/GB8823897D0/en
Priority claimed from GB888823895A external-priority patent/GB8823895D0/en
Priority claimed from GB888823896A external-priority patent/GB8823896D0/en
Application filed by Browning's Clinical Pathology Services Limited filed Critical Browning's Clinical Pathology Services Limited
Publication of WO1989005657A1 publication Critical patent/WO1989005657A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0634—Cells from the blood or the immune system
    • C12N5/0636—T lymphocytes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/46—Cellular immunotherapy
    • A61K39/461—Cellular immunotherapy characterised by the cell type used
    • A61K39/4611—T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/46—Cellular immunotherapy
    • A61K39/464—Cellular immunotherapy characterised by the antigen targeted or presented
    • A61K39/464838—Viral antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2239/00—Indexing codes associated with cellular immunotherapy of group A61K39/46
    • A61K2239/26—Universal/off- the- shelf cellular immunotherapy; Allogenic cells or means to avoid rejection
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/20—Cytokines; Chemokines
    • C12N2501/23—Interleukins [IL]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/20—Cytokines; Chemokines
    • C12N2501/24—Interferons [IFN]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/50—Cell markers; Cell surface determinants
    • C12N2501/59—Lectins

Definitions

  • Lymphokine activation of cells for adoptive immunotherapy eg. of HIV infection.
  • the present invention relates to lymphokine activated cells, therapeutic compositions including such cells, methods of producing lymphokine activated cells and to methods of treatment involving such cells.
  • Adoptive immunotherapy is a process whereby the body's immune system is reinforced by therapeutic means.
  • lymphocytes are withdrawn from a patient, fractionated, purified and then treated with a lymphokine.
  • Lymphocytes activated by treatment with a lymphokine are known as lymphokine activated killer (LAK) cells and are known to have greatly enhanced activity.
  • Suitable lymphokines for use in any aspect of the present invention include alpha interferon, gamma interferon, interleukin 1, interleukin 2 and interleukin 2 produced by phytohaemagglutinin (PHA). After activation the LAK cells are used into the patient.
  • a method for the activation of lymphocyte s from a patient or donor for use in adoptive immunotherapy character ised in that autologous or donor lymphocyte conta ining body liquid is incubated with a lymphok ine ; whereby the resultant culture containing LAK cells is returnable d irec tly to the patient.
  • a second aspect of the invention provides a reinfusable therapeutic composition for use in adoptive immunotherapy which comprises an autologous or donor (allogenic) lymphocyte containing body liquid activated by a lymphokine to provide patient-specific LAK cells.
  • a therapeutic composition for use in adoptive immunotherapy which composition comprises autologous lymphocytes activated with a lymphokine to form lymphokine activated killer cells (LAK cells) for reinfusion into a patient; characterised in that said composition additionally comprises lymphokine activated donor lymphocytes.
  • LAK cells lymphokine activated killer cells
  • a fourth aspect of the invention provides a method of activating donor lymphocytes for use in adoptive immunotherapy, which method comprises, incubating said donor lymphocytes with a lymphokine to give LAK cells, characterised by preparing said LAK cells for direct infusion into a patient.
  • a fifth aspect of the present invention provides a method for activating autologous lymphocytes for use in adoptive immunotherapy, which method comprises incubating said autologous lymphocytes with a lymphokine to give LAK cells; characterised in that donor lymphocytes are incubated with a lymphokine and the admixture of autologous and donor LAK cells is prepared for reinfusion into a patient.
  • lymphocytes are fractionated after incubation with the lymphokine.
  • lymphocytes are collected from a patient to be treated. Such a collection may be by lymphopheresis or, where lymphopheresis is impractical, by straight venesection.
  • lymphokine for example interferon alpha, or gamma, or interleukin 1, or 2 is added to the total collected material, whether this is pheresed cells or whole blood.
  • a lymphokine for example interferon alpha, or gamma, or interleukin 1, or 2 is added to the total collected material, whether this is pheresed cells or whole blood.
  • azidotymidine is added to the lymphocytes to prevent viral replication.
  • the suspension so formed is incubated with the selected lymphokine.
  • the LAK cell containing culture is then immediately reinfused into the patient. Where, however, the patient's lymphocytes are damaged beyond a certain degree, further steps are necessary.
  • the lymphocytes are separated from the mother liquor to give a percentage of lymphocytes of greater than 90% and preferably greater than 95%, having a similar percentage viability.
  • the lymphocytes are then washed with a buffer solution, preferably on a serial basis and are then suspended in a suitable culture medium, for example, RPMI.
  • a suitable culture medium for example, RPMI.
  • the so-formed culture is then incubated for a suitable period, for example, 48 to 72 hours. After incubation, the culture is subjected to centrifugation to separate off the supernatant portion containing the LAK cells for reinfusion. These are purified prior to reinfusion into the patient.
  • donor's cells from a party such as a patient's relative, are formed in precisely analagous way with the exception, of course, that the addition of AZT will not be required. This provides a reservoir of healthy LAK cells which may be utilised to potentiate the activity of autologous LAK cells.
  • the donor LAK cells are added, for example, in an amount of 1-2 to 1:4 to the patient's LAK cells prior to a final purification and preparation step.
  • the present invention relates to a method of treating a patient suffering from an infectious, or oncological disease, comprising infusing LAK cells activated by a method in accordance with any of the first, fourth or fifth aspects of the invention; or with a therapeuctic composition in accordance with the second or third aspect of the invention.
  • the method comprises the step of firstly isolating the cells for activation from the patient to be treated, or another individual.
  • the cells for activation may be a mixture of cells from both said sources.
  • Human immune deficiency virus HIV is the etiological pathogen in the development of the acquired immune deficiency syndrom (AIDS). The virus infects and destroys activated cells of the immune system leading to a profound defect in the host's natural immunity.
  • a vaccine to prevent worldwide spread of the disease is not at present obtainable.
  • the spread of the disease among heterosexual populations is still increasing.
  • treatments that halt the disease, if not to cure it, are urgently required.
  • the virus must bind to a receptor site on the surface of a "host” cell and thereafter enter the cytoplasm of the host cell. The virus then causes the insertion of its "foreign" genetic material into the host cell's genetic material; with the result that, the host cell is caused to produce fresh virus and eventually to perish. Additionally, the normal function of the host cell is often impaired and the host cell may age prematurely.
  • the virus replicates by binding to a CD4 receptor site on the surface of a cell having such a site, for example a T-helper cell (a species of lymphocyte), and, thereafter entering the cell and inserting foreign genetic material into that of the T-helper cell.
  • a T-helper cell a species of lymphocyte
  • HIV sheds its envelope and exposes its core RNA.
  • a portion of DNA, corresponding to the viral RNA, is then inserted into the host cell's DNA by enzymatic action.
  • the host cell's mechanism then begins to produce fresh HIV on the 'instructions' of the DNA introduced by the infecting virus.
  • the fresh HIV is then expressed out through the infected cell's wall and into the circulation, where the fresh virus may infect further cells.
  • the virus is able to remain in the host cell's cytoplasm, prior to shedding its envelope, for a considerable period of time and, hence, effectively lie dormant, safe from antibodies thereto in the circulation. T-helper cells infected in this manner have impaired performance and age prematurely.
  • HIV is able to enter a cell having a CD4 receptor within less than 1/2 hour, which is not sufficient time for the immune system to respond to the presence of the virus and produce antibodies thereto.
  • the infected cells act as reservoirs of virus, although they may be destroyed by the action of the immune system once it has learnt to respond to the virus, when they are actually expressing HIV.
  • the response of the immune system to HIV and other opportunistic infections, which give the characteristic symptoms of AIDS and AIDS related complex (ARC), is impaired because the Helper cell population is reduced, both by the cytopathic effect of HIV and the action of the immune system's killer cells destroying infected helper cells.
  • a composition comprising lymphokine activated cells having binding sites for an infectious agent characterised in that said lymphokine activated cells have been modified after activation so as to be unable to facilitate virus reproduction.
  • infectious agent is a virus, more preferably it is a retro-virus and most preferably HIV.
  • the cells may be fractionated or purified before or after treatment with a lymphokine.
  • the activated cells are lymphocytes. It is believed that when a lymphocyte is activated with a lymphokine, the number of suitable binding sites for HIV on the surface of the resulting lymphokine activated cell, is greatly increased over that on a normal resting lymphocyte. Thus, HIV is more likely to bind with an activated lymphocyte than a normal resting lymphocyte. Accordingly, when introduced into the circulation of an individual infected with HIV, activated lymphocytes bind to free virus in preference to normal non-activated lymphocytes. Since an activated cell in accordance with the present invention cannot reproduce virus, any viral genetic material introduced into such a cell by a virus which has entered that cell, is ineffective. Accordingly, once a virus has entered an activated cell in accordance with the seventh aspect of the present invention, that virus is effectively eliminated from the circulation and prevented from replicating.
  • modification of the activated cells involves the disruption, or destruction of the cells' genetic material.
  • this modification is achieved through the use of a chemical agent such as a DNA inhibitor, which prevents reproduction, but does not interfere with normal cell function.
  • a suitable DNA inhibitor is mitomycin-C.
  • modification may be achieved through the irradiation of the activated cells.
  • Lymphocytes for producing activated cells in accordance with the seventh aspect of the present invention may be derived from a healthy donor, both directly or indirectly, or they may be produced by an established cell line, such as CEM or H9 (these originate from T-Helper cells and are commercially available). Most preferably, the lymphocytes include T-helper cells. Alternatively, the lymphocytes may be derived from cell lines which have been 'improved' by recombinant techniques to have additional receptor sites.
  • the present invention provides a process for activating and modifying cells having binding sites for an infectious agent comprising treating said cells with a lymphokine and, subsequently, modifying the resultant activated cells in order to render virus replication impossible in said cells, while maintaining the cells' ability to bind said infectious agent.
  • the infectious agent is a virus, more preferably it is a retro-virus and most preferably, it is HIV.
  • the cells for modification are lymphocytes.
  • modification of the activated cells involves the disruption or destruction of the cells genetic material.
  • this modification is achieved through the use of a chemical agent, for example a DNA inhibitor such as mitomycin-C, or through the irradiation of the activated cells.
  • lymphocytes for producing the activated cells are derived from a healthy donor either directly or indirectly, or they may be derived from an established cell line, such CEM or H9.
  • the lymphocytes include T-helper cells.
  • the lymphocytes may be derived from cell lines which have been 'improved' by recombinant techniques to have additional receptor sites.
  • a therapeutic composition for use in the treatment of infection with an infectious agent including HIV infection or AIDS, comprising activated cells in accordance with the seventh aspect of the invention, or cells processed in accordance with the eighth aspect of the present invention.
  • a method of treating a human patient infected with an infectious agent comprising infusing a composition in accordance with the ninth aspect of the present invention into said patient.
  • the infectious agent may be a virus, is preferably a retro-virus and more preferably HIV.
  • the ninth aspect of the present invention has the effect of filtering virus from a patient.
  • Activated cells in accordance with the seventh aspect of the present invention only survive for a period of
  • a process for reducing the concentration of an infectious agent such as HIV, or a like virus, in infected plasma comprises, incubating said plasma with a composition in accordance with the seventh aspect of the present invention.
  • activated cells in accordance with the seventh aspect of the present invention are removed from said plasma after binding with HIV, or a like virus. Preferably, this removal is effected by centrifuging the plasma and inventive composition mixture after an incubation period of 1-24 hours. Plasma so treated may be returned to a patient in a form having a much reduced level of HIV and substantially no activated cells, or remains thereof.
  • healthy lymphocytes are stimulated to respond to the presence of an antigen, exemplified by an infectious agent such as virus, when exposed to said antigen.
  • an infectious agent such as virus
  • a person becomes infected with a strain of the common cold virus, his immune system is firstly stimulated by the presence of the virus and, once so stimulated, it responds to the presence of the virus and eliminates it.
  • the virus population expands rapidly, so that the infected individual exhibits the symptoms of a cold; but when the immune system responds, these symptoms recede as the viral population is decimated by the responsive action of the immune system.
  • An individual's immune system may also be taught to respond to the presence of a harmful virus by prior stimulation with incomplete, or dead virus of the same type, or with a virus of a similar, but less harmful type such as a genetically engineered virus.
  • This process is the well known vaccination process and its advantage is that an individual's immune system, after vaccination, will respond rapidly to an invasion by a harmful virus, to eliminate that virus before it has a chance of multiplying sufficiently to cause harm. Otherwise, without vaccination, a harmful virus can multiply sufficiently to harm, or even kill, before an infected individual's immune system has time to be stimulated and to respond. Vaccination also prevents secondary infection with the same virus and thus gives long lasting protection.
  • a process for activating and stimulating lymphocytes to respond to an infectious agent comprising incubating lymphocytes with a lymphokine to produce LAK cells and, exposing the lymphocytes to a suitable antigen of said infectious agent.
  • the lymphocytes are exposed to the antigen after activation and, more preferably the antigen is an infectious agent such as a virus or a retro-virus; the virus or retro-virus may be alive, dead, or in some way disabled; the antigen may also be a virus particle or a protein.
  • the lymphocytes for treatment may be derived from a healthy donor, not infected by said infectious agent, or from a suitable cell line.
  • the lymphocytes for activation are exposed to the antigen prior to activation, to thereby become capable of controlling said agent.
  • activated and stimulated lymphocytes produced by a process in accordance with the present invention may be infused into a patient infected with the infectious agent, wherein the stimulated LAK cells act to reduce the level of infection within the patient.
  • Such an infusion is particularly effective where the infectious agent has a harmful or debilitating effect on a patient who alone, is unable to react sufficiently to the presence of the infectious agent.
  • the infectious agent is HIV and the antigen is live HIV, inactive HIV, or a portion thereof.
  • the use of lymphocytes, stimulated and activated in vitro by a method of the present invention to react to HIV, is particularly useful in the treatment of AIDS.
  • An AIDS sufferer's own immune system is damaged through Helper cell depletion and is unable to reduce the level of HIV infection to thereby recover some of its function.
  • lymphocytes processed in accordance with the present invention to respond to the HIV strain infecting a particular AIDS sufferer are infused into the sufferer, the level of HIV infection may be reduced by the activity of the stimulated and activated lymphocytes.
  • the lymphocytes for activation are derived directly or indirectly from a human or animal infected with said infectious agent, wherein said human or animal exhibits little or none of the deleterious symptoms of infection.
  • the infectious agent may be a virus, is preferably a retro-virus and most preferably HIV.
  • the present invention provides antibodies capable of controlling an infectious agent by neutralisation, which antibodies are harvested from lymphocytes previously exposed to an infectious agent and capable of controlling said agent.
  • Antibodies in accordance with the thirteenth aspect of the present invention may be employed in the same manner as LAK cells in accordance with the twelfth aspect of the present invention, in order to alleviate the symptoms of a patient suffering from the effects of infection with an infectious agent.
  • the antibodies are derived either directly or indirectly from a human or animal infected with said agent, but exhibiting little or none of the deleterious symptoms of infection.
  • the infectious agent may be a virus, is preferably a retro-virus and most preferably HIV.
  • the present invention provides a therapeutic composition, comprising cells in accordance with the twelfth aspect of the invention, antibodies in accordance with the thirteenth aspect of the invention, or a mixture of said cells and said antibodies.
  • a therapeutic composition in accordance with the present invention is to be useful in treating a particular patient, for the lymphocytes used to have been exposed to the same infectious agent as that afflicting the patient, or for the antibodies to be specific for this agent.
  • the lymphocytes in treating AIDS or HIV infection, the lymphocytes must have been exposed to the same strain of the virus as that which afflicts the patient, or the antibodies must be specific for said strain. This is easily achieved by obtaining lymphocytes or antibodies, for activation, from an HIV positive individual who is without AIDS symptoms and, who is infected with the same strain of HIV as the AIDS sufferer it is intended to treat.
  • lymphocytes or antibodies, for activation may be obtained from serum derived from the sexual partner of the patient, or from a parent to treat a child.
  • the serum may be drawn from an individual during an early phase of HIV infection, or whenever an individual is not exhibiting AIDS symptoms. and used to provide LAK cells, or activated antibodies for treating the same individual, when he is exhibiting AIDS symptoms.
  • the serum may be stored in a frozen state until it is required.
  • the stimulated and activated lymphocytes act both directly on the virus by destroying lymphocytes (helper cells) which are expressing HIV and also, by activating the cytotoxic cells within the patient's circulation to attack both opportunistic infections and HIV expressing lymphocytes.
  • helper cells lymphocytes
  • the cytotoxic cells (CD8) of an infected individual are not damaged or infected by HIV, because they lack CD4 receptor sites, but they require activation by Helper cells before being able to respond to the presence of infection. It is for this reason that the cytotoxic cells of a person with AIDS do not respond to HIV or other opportunistic infections.
  • the sufferer's immune sytem recovers some of its function, thereby alleviating some AIDS symptoms.
  • the antigen is derived from serum extracted from a patient infected with the infectious agent it is intended the activated lymphocytes should respond to.
  • the serum is extracted from the patient it is intended to treat and said serum includes the virus HIV.
  • the serum may be extracted from the patient and HIV isolated from a culture of lymphocytes derived from the serum and thereafter used to stimulate donor lymphocytes.
  • the HIV is exposed to ultraviolet light for between 24 and 48 hours to inactivate the virus.
  • the virus may be heated at 56°C for 1/2 hour.
  • the present invention provides a process for activating lymphocytes which have been exposed to an infectious agent comprising incubating the lymphocytes with a lymphokine to produce LAK cells.
  • the infectious agent is a retro-virus, such as HIV.
  • the lymphocytes for activation are derived from an individual infected with HIV and, during activation, the lymphocytes are treated with a drug which inhibits viral replication, such as AZT.
  • the present invention provides activated and stimulated lymphocytes produced by a process in accordance with the twelfth or fifteenth aspects of the present invention.
  • the present invention provides therapeutic compositions comprising activated and stimulated lymphocytes produced by a process in accordance with the twelfth or fifteenth aspects of the present invention.
  • the genetic material within the lymphocytes is modified after activation so as to render virus production by the lymphocytes impossible. This may be achieved by treating the LAK cells of the invention with a suitable chemical composition such as a DNA inhibitor or with radiation.
  • a suitable DNA inhibitor is the drug mitomycin-C.
  • the present invention provides a method of producing LAK cells comprising obtaining lymphocytes, either directly or indirectly, from an individual infected with an infectious agent who exhibits substantially no deleterious symptoms of such infection, and activating said lymphocytes by incubation with a suitable lymphokine.
  • infectious agent is a virus, preferably a retro-virus and more preferably HIV.
  • the lymphocytes or antibodies are treated with a substance to remove substantially all free infectious agent, or to protect activated cells from infection with the infectious agent.
  • the substance employed may be AZT (azidothymidine) or dextran sulphate.
  • the present invention relates to a method of treating an individual infected with an infectious agent comprising, administering lymphocytes activated and stimulated to respond to said infectious agent in accordance with the present invention.
  • donor LAK cells promote increased suppressor cell activity, which in turn casues a patient's ability to respond to foreign cells to be reduced. Nevertheless the donor LAK cells maintain an ability to kill the patient's malignant, or virus infected cells.
  • the twentieth aspect of the present invention provides a composition for use in a method of increasing suppressor cell activity, which composition comprises activated lymphocytes from a patient, or donor prepared by incubating a body liquid containing autologous or donor lymphocytes with a lymphokine.
  • composition of the invention may be used to suppress patients' ability to respond to foreign cells and to thereby induce a tolerance to the presence of foreign tissue.
  • composition of the twentieth aspect of the present invention is usable in transplant procedures in order to reduce the incidence of rejection of transplanted tissue. This usefulness applies to the transplantation of both heamopoietic tissue and other organs such as kidneys, livers, and hearts.
  • Autologous lymphocytes were collected from a patient by lymphopheresis using a cell separator to give between 2.5 and 10 ⁇ 10 9 /l.
  • the volume of fluid collected during pheresis varies between 200 and 400 ml, the proportion of lymphocytes varying between 55 and 70% since, of course, other cells are collected by this process.
  • 500 ml of blood may be taken from the patient by straight venesection.
  • the suspension is now incubated for one hour at about 37oC, without antibiotic or medium added.
  • the process is conducted in 1 litre transfer packs produced by "Fenwall”. In fact only 3-500 ml of the suspended cells are placed in each 1 litre bag. After the addition of Interferon Alpha and incubation for one hour, the entire material is reinfused into the patient.
  • lymphocyte population of lymphocytes
  • lymphocytes are then separated by Ficoll-Paque to give an end percentage of lymphocytes of greater than 95 % having a viability of greater than 95%.
  • the separated lymphocytes are then washed with Hanks Balanced salt solution and subsequently suspended in a culture medium RPMI 1640 supplemented with 5-10% of autologous plasma containing INF and a suitable antibiotic, for example streptomycin and/or penicillin; 100 units of each.
  • lymphocytes are suspended in the RPMI culture
  • the cells are centrifuged at 350 g force for 10 minutes and the consequent pellets are then collected, washed three times with Hanks Balanced Salt Solution which is supplemented with 5% autologous patients' plasma and 10 units of preservative-free heparin.
  • the cells are finally suspended in 0.9 sodium chloride, 10 units of preservative-free haparin and 10% autologous plasma making the final volume of pre-infusion composition 200 ml.
  • the volume of cells reinfused into the patient vaies between 0.3-7 ⁇ 10 9 /1.
  • donor cells are extracted from a healthy individual, for example, a family member of the patient.
  • the donor cells are treated in precisely analagous form as the patient's cells with the exception, of course, that since the cells are healthy, there is no need to add AZT.
  • the donor cells are centrifuged in precisely the same way as the autologous cells, to provide healthy LAK cells containg the identifyable lymphokines, Interleukin 2 , Interleukin 1 and Gamma Interferon. Other lymphokines are also present but these have not yet been identified.
  • the donor lymphocytes used for lymphokine production may be obtained by centrifugation of 50% of the culture medium at 24, 48 and 72 hours respectively.
  • Such donor LAK cells may be stored at -20 C or may be infused directly into the patient or added directly to the patient's cultured LAK cells in a volume of 1:4 or 1:2. This suspension may be directly added to the autologous LAK cells at any convenient period.
  • lymphokines from the donor LAK cells may be extracted and used to stimulate autologous lymphocytes.
  • the invention provides a method for activating LAK cells without fractionation, and a method for potentiating the action of LAK cells by addition thereto of donor LAK cells as defined.
  • Allogeneic lymphocytes were administered to patients suffering from malignant disease and HIV infection.
  • the donors were variously spouses and blood relatives.
  • In only one case was the donor HLA, MLR an blood group identical to the patient.
  • the lymphocytes were obtained by a process of lymphocytopheresis and further purified using ficol-paque. Activation of the lymphocytes was achieved by adding Interferon Alpha at 1000 units per millilitre. The activated lymphocytes were then incubated for between one and 72 hours and, on occasio even longer. The activated lymphocytes were then administered to the patients. The incidence of fever was less than 10% of reinfusions. This is more or les the same incidence of fever as seen in reinfusion of autologous cells. No other side effect has been identified. In some cases, anti donor lymphocyte antibodies were identified after treatment, while in other no such antibodies were seen. It is considered that the patients had a prior degree of immune suppression because of their disease. In some cases the absolute lymphocyte numbers were normal.
  • Autologous lymphocytes were collected from a patient lymphopheresis using a cell separate to give between 2.5. and 10 ⁇ 10 9 /1.
  • the volume of fluid collected during pheresis varies between 200 and 400 ml, the proportion of lymphocytes varying between 55 and 70% since, of course, other cells are collected by this process.
  • 500 ml of blood may be taken from the patient by straight venesection.
  • the suspension is now incubated for one hour at about 37°C.
  • the resulting activated cells were then treated with mitomycin-C at a concentration of 25-40 /ig/ml .
  • the cells were incubated with the mitomycin-C for 45 minutes at 37oC.
  • the modified activated cells were then washed 2-3 time and were prepared for infusion into an individual infected with HIV in 200 ml saline, supplemented with 10% autologous plasma.
  • the lymphocytes may be modified through irradiation with between 300 and 3000 rads, in place of exposure to mitomycin-C.
  • Lymphocytes were collected from the sexual partner of a patient suffering from AIDS, by lymphopheresis using a
  • lymphocyte separator to give between 2.5 and 10 ⁇ 10 9 lymphocytes/Litre.
  • lymphocyte collection Prior to lymphocyte collection, it is established that the sexual partner, although not suffering from the symptoms of AIDS, is infected with HIV.
  • the volume of fluid collected during pheresis varies between 200 and 400 ml, the proportion of lymphocytes varying between 55 and 70* since, of course, other cells are collected by this process.
  • 500 ml of blood may be taken from the partner by straight venesection.
  • 1000 IU alpha interferon is added togethe with 1-5 micromoles of AZT. This suspension was then incubated for one hour at about 37oC. The process is conducted in a one litre transfer pack produced by
  • the entire material is infused into the patient suffering from AIDS.
  • buffy coat comprised approximately 8-18 ⁇ 10 9 lymphocytes per Litre.
  • the buffy coat was activated with 1000 IU of interfer alpha (IFN) plus 10/umol of AZT. After 1 hour's incubation, the plasma was removed and the lymphocyte separated on ficoll and then cultured with 10 ⁇ mol of AZT and 1000 IU of IFN, in a mixture of the previousl isolated plasma and fresh donor plasma. This mixture was cultured for 3-5 days and then washed once before suspension in the husband's plasma and 200 ml of saline.
  • IFN interfer alpha
  • the patient was infused with this suspension and the treatment repeated five times.
  • the subjects were a mother and child, the child suffering from AIDS and the mother being HIV positive. After treatment, the patient ' s AIDS symptoms, caused b y opportunist infections were considerably reduced and, in some cases, completely eliminated.
  • the subjects were a mother and ch ild, the child suffering from AIDS and the mother being HIV posit ive.
  • Lymphocytes were collected from the mother in a buffy coat derived by lymphopheresis , using a cell separator.
  • Serum is drawn from a patient suffering from AIDS, lymphocytes from the serum are cultured and HIV is isolated from that culture. The HIV so isolated is then exposed to ultraviolet light for 24-48 hours to inactivate the virus. Alternatively, the virus may be inactivated by heating, or used in its live state.
  • Donor Lymphocytes are collected from a healthy individual by lymphopheresis using a cell separator to
  • lymphocyte containing body liquids 1000 IU of alpha interferon is added.
  • the suspension is then incubated for three days in order to stimulate the lymphocytes.
  • Live or inactivated virus derived in the manner set out above, is then added to the stimulted lymphocytes and the resulting mixture is incubated for a further five days.
  • the mixture of activated lymphocytes and live or inactivated virus is treated with AZT, or another antiviral drug, in order to protect activated lymphocytes from infection with virus. The mixture is then tested for cytotoxic and viral inhibitory effects before use.
  • the stimulated and activated lypmohcytes may be infused into the patient from whom the HIV was originally derived, in order to reduce the amount of HIV in his circulation and to combat the effects, both directly and indirectly, of opportunistic infections.
  • lymphopheresis The total cell count was 8-13 ⁇ 10 9 cells per litre.
  • the buffy coat, so derived, was treated with 1000 IU's of interferon alpha (IFN) and 10 ⁇ mol of AZT. The resulting mixture was incubated for 1 hour and, thereafter, the supernatant was removed.
  • IFN interferon alpha
  • lymphocytes were then separated out from the remainder of the mixture on ficoll and thereafter cultured with 10 fl mol of AZT, 1000 IU of IFN in a mixture comprising the supernatent derived as set out above plus 25% of a similar supernatant, derived from healthy donor culture.
  • the resulting mixture was suspended in 200 ml of untreated autologous plasma and saline, and treated with 10 ⁇ mol of AZT for infusion into the patient suffering from ARC.
  • the patient was given a total of 10 such infusions and, his symptoms of ARC began to improve after the eighth infusion.
  • lymphocytes Culture the separated lymphocytes in a fenwall culture bag in the presence of 10 ⁇ mol of AZT, preparation A and 25% of the supernatant from a lymphokine activated culture obtained from a healthy donor. This mixture should be cultured for 3-7 days. 6. After culturing, the lymphocytes should be was with Hanks solution and resuspended in untreated fres autologous plasma and saline at a total volume of 200 ml.
  • the suspension Prior to infusion, the suspension is tested for viability, infection and functional activity against various target infections.
  • the suspension is then reinfused into the patient, preferably 8-12 times. On infusion, the suspension comprises 1-10 ⁇ 10 9 cells per litre.
  • PBL peripheral blood lymphocyte
  • Acid citrate dextrose was used as the anti-coagulant.
  • Autologous Lymphocytes were harvested in 5-6 surges over a period of approximately 2 hours at any one collection. HIV+ and
  • HIV- donor lymphocytes were similarly harvested.
  • the total cell collection was on average 9 ⁇ 10 9 /L.
  • the volume collected varied between 250 and 400 ml.
  • Lymphocytes were harvested variously on 5-13 occasions at usually 3-day intervals.
  • Lymphocytes were separated on Ficoll Hypaque and washed with Hanks Balanced Salt Solution (HBSS) free of calcium and magnesium. The gradient was centrifuged at HBSS.
  • HBSS Hanks Balanced Salt Solution
  • Lymphocytes at the plasma Ficoll Hypaque interface were collected and washed in HBSS three times. Ficoll separation and cell harvesting were performed under sterile conditions. Lymphocytes were cultured in RPMI 1640 supplemented with 10* autologous plasma. Interferon was added at a final concentration of 100-1000 IU/ml. The cells were cultured in the presence of AZT in a concentration of 10 ⁇ . mols for 3-10 days. The cells were washed in HBSS and resuspended in 200 ml of normal saline with 10% plasma, prior to infusion or reinfusion into the patients.
  • PHA phytohaemagglutinin
  • IL2 interleukin 2
  • the final cell concentration of 1 ⁇ 10 6 /ml was cultured in plastic Petri dishes over 1 hour at 37oC. Adherent cells were left behind in the Petri dish. Further incubation of the cells took place in 5 ml syringes packed with nylon fiber. After 1 hour's incubation at 37oC. the non-B cells were eluted with RPMI 1640 . The eluted cells were then treated with monoclonal antibody to CD8 and complement. The purification of CD4 cells was of the order of 85%.
  • the purified population of CD4 cells was mixed with
  • HIV+ plasma to give a concentration of 1 ⁇ 10 6 /ml.
  • the culture was further diluted with an equal volume of RPMI.
  • the cells were cultured in tissue culture flasks for 7 days at 37 oC. in the presence of 100 i.u./ml of IL2.
  • the infected cell collection was used as a target for cytotoxic T cells in a short 51 Cr release assay
  • HIV antibody and antigen were assessed by end point titration methods with Eliza (Abbott).
  • End point titration methods with Eliza (Abbott).
  • the supernatant from infected cultures was introduced into non-infected lymphocytes in culture.
  • Target cell line (K562) was labelled with 100 microcuries of sodium chromate. The cells were incubated at 37 oC. for 45 minutes. After incubation the cells were then washed 3 times and re-suspended in
  • Autologous blast target cells were generated by incubation of the patient's blood lymphocytes in the presence of PHA 10/fg/ml and interleukin 2, 1000 IU/ml for 72 hours.
  • the cells were infected with HIV on Day 3 and Day 5 and labelled with 51 Cr sodium chromate as above. The infected cells were tested for functional activity against target cells.
  • CD4 and CD8 numbers were determined using monoclonal antibody conjugated to fluorescein by a technique of indirect immunafluores ⁇ ence.
  • LAK lymphokine activated killer
  • lymphocytes were treated in vitro variously with interferon (IFN ), interleukin 2 (IL2), azidothymidine (AZT) and mitomycin-C.
  • IFN interferon
  • IL2 interleukin 2
  • AKT azidothymidine
  • mitomycin-C mitomycin-C
  • Protocol selection was dependent upon the patient's condition and disease stage at presentation.
  • Protocol 1 (autologous cells) was used for the treatment of patients with ARC. These patients had white cell counts >4 ⁇ 10 9 /L and CD4 > 0.5 ⁇ 10 9 /L.
  • Protocol 2 (HIV+ allogeneic cells) was used in the treatment of a patient with AIDS.
  • the white cell count was ⁇ 4 ⁇ 109/L, CD 4 > 0.04 ⁇ 109 /L and CD 8 ⁇ 0.8 ⁇ 10 9 /L.
  • the patient selected for this protocol was infected with the same strain of HIV as her husband who was used as the donor.
  • This protocol could be used in vertically transmitted infection in addition to horizontally transmitted disease.
  • the donor was asymptomatic.
  • the purpose of this protocol was to overcome the problem of very low numbers of CD4 cells in stage 5 and 6 disease. Furthermore, patients with late stage disease, beacuse of leukopenia and debility, make them poor candidates for leukapheresis.
  • Protocol 3 allogeneic activated cells treated with mitomycin-C was used in AIDS sufferers, the donors being healthy individuals. Cells for infusion were treated with mitomycin-C to prevent virus infection and replication in donor cells. Although DNA damage was to be anticipated, it was considered that the cells would retain the ability to bind the free HIV antigen and that the release of CD4 signals would remain intact.
  • Table 1 sets out the clinical features of the six subject patients
  • Table 2 shows immunoconversion of all six subject patients before and after treatment
  • Table 3 shows the cytotoxicity induced by LAK cells from PBL of patients 1 and 2, before treatment and after 2-4 treatments;
  • Table 4 shows the lymphocyte count for patient 6 during treatment
  • Table 5 sets out the cell viability for patients 1-4 and 6 before treatment and after 5 infusions and;
  • Table 6 sets out the antigen titer before, during and after treatment for patients 3-5.
  • Figure 1 shows the rate of syn ⁇ ytium formation in cultures taken from patients 1-3 before treatment
  • Figure 2 shows the increase in the number of syncytia present in a 4 day old culture taken from patients 1-3 prior to reinfusions 1-4;
  • FIG. 3 shows the T-cell subsets in patient 4 before treatment and after 1-10 treatments
  • FIG. 4 shows the T-cell subsets in patient 3 after treatment with HIV+ and normal allogenic activated cells
  • FIG. 5 shows the lymphocyte subsets after treatment with eight infusions (Patient 2);
  • HIV+ cells express the envelope glycoprotein GP120 which produces cell fusion and sincytium formation of both infected and non-infected cells. Ultimately, cell death occurs among all cells bearing CD4 molecules.
  • mononuclear cells obtained by pheresis and cultured in the presence of interferon 100-1,000 i.u. /ml. and AZT 5/ ⁇ mols were clumped together to form nucleated giant cells which reached a peak of 65-70% of cells in culture on day 4 in patients 1 and 2.
  • patient 3 had little increase in sincytail formation in culture. See figures 1 and 2.
  • the differences between patients 1 and 2 and patient 3 may be attributable to the concentrations of the AZT used.
  • patients 1 and 2 may have reached the more advanced stage of the disease. Furthermore, variation in virulence of the strains of virus among these patients could not be excluded.
  • HIV antigenaemia was identified in the three AIDS patients before treatment with activated cells, while the three ARC patients were HIV antigen negative. In two of the ARC patients tested one year after the initial treatment, antigen negative status was maintained. See table 2.
  • Antigen was detected at a level of 1 in 16 in patient 4 before treatment with activated cells. After one reinfusion, antigen was not detected and this remained so until the end of treatment.
  • Patient 5 had an antigen titre of 1 in 8 at presentation. This reduced to 1 in 2 after one infusion, and this patient remained antigen positive throughout treatment.
  • Patient 6 was antigen positive and remained antigen positive when treatment was aborted after three reinfusions. See table 6. Culture of isolates from plasma and lymphocytes of patients 4 and 5 proved to be highly infectious with the death of about 70% of the cultured cells after five days. The reduction of cell death in culture shown in these patients after treatment suggests a reduction in the level of virion. See table 5.
  • Cytotoxic NK normal killer
  • T-cell activity was improved following treatment (Table 3). This was identified using K562 targets where, in patient 1, post treatment cytoxicity was 35% higher than pre treatment and 23% higher in patient 2. A similar effect was seen with HIV infected blasts where the increase was 29% for patient 1 and 17% for patient 2.
  • the HIV virus particularly infect CD4 cells.
  • the infected cells express the HIV envelope glycoprotein GP120, which induces cell fusion, sincytium formation and ultimately cell death.
  • the initially high level of cell death and sincytium formation was shown to be reduced in vitro in the presence of AZT and interferon.
  • interleukin 2 The addition of interleukin 2 to interferon produced a striking augmentation in the numbers of NK cells and
  • CD8 cells were capable of inducing lysis in Nk sensitive targets (K562) and HIV infected targets. Cytotoxicity was increased to twice that (see treatment table). The combination of interferon , interleukin 2 and AZT in vitro suggests that this combination will be relevant to in vivo management. CD8 numbers were increased in the circulation following treatment (figure 5 and table 4). The enhancement in cytotoxicity which we witnessed could be due to this increase, and this may play a significant role in preventing HIV infection of other cells. It is probable that the CD8 cells kill virally infected cells and neutralise free antigens.
  • P24 antigen has been used in other clinical studies to monitor the effect of HIV inhibitory agents.
  • the level of antigen in patients 4 and 5 dropped from 240 to 90 picograms/L in less than 10 days and following 1 reinfusion.
  • Clinical studies with AZT have shown the same reduction in antigen level, but only after 16 weeks.

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Abstract

La présente invention se rapporte à des cellules activées par lymphokine, à des préparations thérapeutiques contenant de telles cellules, à des procédés de production de cellules activées par lymphokine et à des procédés de traitement utilisant de telles cellules. Les cellules activées sont utiles dans des procédés de traitement de maladies infectieuses, telles que le SIDA et l'ARC.
PCT/GB1988/001134 1987-12-17 1988-12-19 Activation de cellules par lymphokine pour l'immunotherapie adoptive, par exemple de l'infection par l'hiv WO1989005657A1 (fr)

Applications Claiming Priority (10)

Application Number Priority Date Filing Date Title
GB878729410A GB8729410D0 (en) 1987-12-17 1987-12-17 Method for production of lymphokine activated killer(lak)cells & lak cells so produced
GB8729410 1987-12-17
GB888817082A GB8817082D0 (en) 1988-07-18 1988-07-18 Method for production of lymphokine activated killer(lak)cells so produced & their use
GB8817082.4 1988-07-18
GB888823897A GB8823897D0 (en) 1988-10-12 1988-10-12 Process for stimulating & activating lymphocytes
GB888823895A GB8823895D0 (en) 1988-10-12 1988-10-12 Lymphokine activation of lymphocytes
GB8823896.9 1988-10-12
GB8823895.1 1988-10-12
GB8823897.7 1988-10-12
GB888823896A GB8823896D0 (en) 1988-10-12 1988-10-12 Improvements relating to lymphokine activated cells

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Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0584715A2 (fr) * 1992-08-26 1994-03-02 Dr. Kübler GmbH Isolation et cultivation de cellules transformées et procédure de préparation des anticorps contre ces cellules
US5316763A (en) * 1991-07-10 1994-05-31 The United States Of America As Represented By The Department Of Health And Human Services Short-term anti-CD3 stimulation of lymphocytes to increase their in vivo acitivity
US5607917A (en) * 1991-12-31 1997-03-04 Zymogenetics, Inc. Methods for reducing blood loss
US5641677A (en) * 1991-04-05 1997-06-24 Regents Of The University Of Minnesota Method of enhancing the immunotherapeutic activity of immune cells by depletion of CD8+ T cells
US5650152A (en) * 1988-10-27 1997-07-22 Regents Of The University Of Minnesota Liposome immunoadjuvants containing 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
EP1401495A2 (fr) * 2001-06-01 2004-03-31 Xcyte Therapies, Inc. Reparation et regeneration de tissus induites par des cellules t
WO2005051927A1 (fr) * 2003-11-26 2005-06-09 Kureha Corporation Procede de culture de lymphocytes t cd4 positifs par l'intermediaire d'une culture de vih-1, et inhibiteur de croissance du vih-1
WO2006105219A2 (fr) * 2005-03-28 2006-10-05 Xcyte Therapies, Inc. Therapie faisant appel aux lymphocytes t pour le traitement de la cachexie et de maladies chroniques

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2106935A (en) * 1981-10-01 1983-04-20 Otsuka Pharma Co Ltd Cancer cell-combatting lymphocytes process, for the production thereof, and anticancer agents containing said lymphocytes
WO1987000054A1 (fr) * 1985-06-28 1987-01-15 The Regents Of The University Of Minnesota Procede et composition pour le traitement du cancer et de tumeur s non malignes
EP0211769A2 (fr) * 1985-08-08 1987-02-25 THE UNITED STATES OF AMERICA as represented by the Secretary United States Department of Commerce Immunothérapie adoptive comme mode de traitement chez les humains
WO1987006610A1 (fr) * 1986-04-28 1987-11-05 Endotronics, Inc. Procede de culture de leucocytes
WO1988000970A2 (fr) * 1986-08-08 1988-02-11 Regents Of The University Of Minnesota Procede de culture de leucocytes
EP0257962A2 (fr) * 1986-08-29 1988-03-02 Becton, Dickinson and Company Méthode de traitement de sang périphérique pour améliorer l'immunothérapie par cellule tueuse activée par une lymphokine
EP0260714A2 (fr) * 1986-09-19 1988-03-23 Oncogen Limited Partnership Utilisation de lymphocytes T actives dans la préparation d'un médicament pour le traitement du SIDA
WO1988007077A1 (fr) * 1987-03-11 1988-09-22 The Children's Hospital, Incorporated Procede de production de lignees de lymphocytes t a specificite antigenique et utilisation therapeutique

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2106935A (en) * 1981-10-01 1983-04-20 Otsuka Pharma Co Ltd Cancer cell-combatting lymphocytes process, for the production thereof, and anticancer agents containing said lymphocytes
WO1987000054A1 (fr) * 1985-06-28 1987-01-15 The Regents Of The University Of Minnesota Procede et composition pour le traitement du cancer et de tumeur s non malignes
EP0211769A2 (fr) * 1985-08-08 1987-02-25 THE UNITED STATES OF AMERICA as represented by the Secretary United States Department of Commerce Immunothérapie adoptive comme mode de traitement chez les humains
WO1987006610A1 (fr) * 1986-04-28 1987-11-05 Endotronics, Inc. Procede de culture de leucocytes
WO1988000970A2 (fr) * 1986-08-08 1988-02-11 Regents Of The University Of Minnesota Procede de culture de leucocytes
EP0257962A2 (fr) * 1986-08-29 1988-03-02 Becton, Dickinson and Company Méthode de traitement de sang périphérique pour améliorer l'immunothérapie par cellule tueuse activée par une lymphokine
EP0260714A2 (fr) * 1986-09-19 1988-03-23 Oncogen Limited Partnership Utilisation de lymphocytes T actives dans la préparation d'un médicament pour le traitement du SIDA
WO1988007077A1 (fr) * 1987-03-11 1988-09-22 The Children's Hospital, Incorporated Procede de production de lignees de lymphocytes t a specificite antigenique et utilisation therapeutique

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
BIKEN JOURNAL, Vol. 30, 1987, Y. KIMOTO et al., "Adoptive Immunotherapy of Malignant Diseases with IL-2-Activated Lymphocytes", pages 29-38. *
CANCER RESEARCH, Vol. 46, June 1986, K. ITOH et al., "Interleukin 2 Activation of Cytotoxic T-Lymphocytes Infiltrating Into Human Metastatic Melanomas", pages 3011-3017. *
DIALOG INFORMATION SERVICES, File 154, Medline 81-89/MAR, Accession No. 05896715, RUSCETTI F.H. et al., "Analysis of Effector Mechanisms Against HTLV-I- and HTLV-III/LAV-Infected Lymphoid Cells"; & J. IMMUNOL., 15 May 1986, 136(10), p3619-24. *
DIALOG INFORMATION SERVICES, File 154, Medline 81-89/MAR, Accession No. 06415462, FROELICH C.J. et al., "Lysis of Human T Cell Leukemia Virus Infected T and B Lymphoid Cells by Interleukin 2-Activated Killer Cells"; & J. IMMUNOL., 1 Dec. 1987, 139(11), p3637-43. *

Cited By (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5650152A (en) * 1988-10-27 1997-07-22 Regents Of The University Of Minnesota Liposome immunoadjuvants containing IL-2
US5773006A (en) * 1988-10-27 1998-06-30 Regents Of The University Of Minnesota Lipsome containing 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
US5641677A (en) * 1991-04-05 1997-06-24 Regents Of The University Of Minnesota Method of enhancing the immunotherapeutic activity of immune cells by depletion of CD8+ T cells
US5316763A (en) * 1991-07-10 1994-05-31 The United States Of America As Represented By The Department Of Health And Human Services Short-term anti-CD3 stimulation of lymphocytes to increase their in vivo acitivity
US5607917A (en) * 1991-12-31 1997-03-04 Zymogenetics, Inc. Methods for reducing blood loss
US5529903A (en) * 1992-08-26 1996-06-25 Dr. Ulrich Kubler GmbH Extraction and cultivation of transformed cells and production of antibodies directed against them
EP0584715A2 (fr) * 1992-08-26 1994-03-02 Dr. Kübler GmbH Isolation et cultivation de cellules transformées et procédure de préparation des anticorps contre ces cellules
EP0584715A3 (fr) * 1992-08-26 1995-02-22 Kuebler Gmbh Dr Isolation et cultivation de cellules transformées et procédure de préparation des anticorps contre ces cellules.
EP1401495A2 (fr) * 2001-06-01 2004-03-31 Xcyte Therapies, Inc. Reparation et regeneration de tissus induites par des cellules t
EP1401495A4 (fr) * 2001-06-01 2005-11-23 Xcyte Therapies Inc Reparation et regeneration de tissus induites par des cellules t
WO2005051927A1 (fr) * 2003-11-26 2005-06-09 Kureha Corporation Procede de culture de lymphocytes t cd4 positifs par l'intermediaire d'une culture de vih-1, et inhibiteur de croissance du vih-1
WO2006105219A2 (fr) * 2005-03-28 2006-10-05 Xcyte Therapies, Inc. Therapie faisant appel aux lymphocytes t pour le traitement de la cachexie et de maladies chroniques
WO2006105219A3 (fr) * 2005-03-28 2007-01-25 Xcyte Therapies Inc Therapie faisant appel aux lymphocytes t pour le traitement de la cachexie et de maladies chroniques

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