WO1997009065A1 - Remede contre maladies auto-immunes - Google Patents

Remede contre maladies auto-immunes Download PDF

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Publication number
WO1997009065A1
WO1997009065A1 PCT/JP1995/001778 JP9501778W WO9709065A1 WO 1997009065 A1 WO1997009065 A1 WO 1997009065A1 JP 9501778 W JP9501778 W JP 9501778W WO 9709065 A1 WO9709065 A1 WO 9709065A1
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Prior art keywords
antibody
mouse
cells
antigen
human
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PCT/JP1995/001778
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English (en)
Japanese (ja)
Inventor
Yoshiko Nishimura
Shin Yonehara
Masato Nose
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Japan Tobacco Inc.
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Priority to PCT/JP1995/001778 priority Critical patent/WO1997009065A1/fr
Publication of WO1997009065A1 publication Critical patent/WO1997009065A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2878—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the present invention relates to a pharmaceutical composition for treating an autoimmune disease, comprising, as an active ingredient, an antibody reactive with a Fas antigen or a part of the antibody reactive with a Fas antigen.
  • the turnover of the cells constituting the living organism must be controlled normally.
  • cells that constitute a living body properly differentiate and proliferate (regeneration of cells), while cells that have become old and have completed a given role must be eliminated.
  • cells that are unnecessary or have a bad effect on the living body resulting from differentiation and proliferation must be immediately killed and removed from the living body.
  • Cell death that occurs in living organisms can be roughly divided into two types.
  • One is cell circulation caused by abnormalities in the external environment surrounding cells, such as heat, infection with viruses, nutritional deficiencies, or tissue damage.
  • Necrosis is the death of cells, and the other is spontaneous death that occurs independently of such abnormalities in the external environment and abnormalities in the external environment. This is the cell death that is inherently called programmed cell death or apoptosis.
  • Fas antigen which is considered to be one of the molecules that induce apoptosis (transmits apoptotic signals into cells) — 1 antigen, called CD 9 5)
  • CD 9 1 antigen
  • Cell surface antigens are attracting attention [Human Fas antigen: Cell, Vol. 66, pp. 233-243, 1991; Journal of Perimental Remedy (J. Exp. Med) 169, 1747-1756, 1989; J. Biol. Chem., 267, 10709-10715, 1992; Mouse Fas. Antigen: Journal of Ibonyology, Vol. 148, No. 4, No. 1274-: 127, 1992, etc.].
  • FasL Fas ligand
  • the Fas antigen is usually a membrane-bound protein that penetrates the cell membrane, but a secreted (soluble) Fas antigen (Fas ⁇ ) lacking the transmembrane region has also been reported. It has also been suggested that the type Fas antigen may inhibit the contact between the membrane-bound type Fas antigen and the Fas ligand, and may inhibit the transmission of apoptotic signals from the Fas antigen [Ayumi Ayumi 170, No. 2, 131-134, 1994; Science, 263, 1759-1762, 1994, etc.].
  • Fas antigen is expressed not only on lymphoid cells such as thymocytes, T cells, cytotoxic T cells (Cytotoxic T cells), B cells or NK cells (Natural Killer cells), but also on cells of non-lymphoid tissues.
  • lymphoid cells such as thymocytes, T cells, cytotoxic T cells (Cytotoxic T cells), B cells or NK cells (Natural Killer cells)
  • various diseases such as viral diseases, inflammatory diseases, cancers or immune system diseases are mediated through the Fas antigen.
  • the association with apoptosis was suggested.
  • the presence of many Fas antigens in lymphocyte cells has led to a possible link between immune system diseases and apoptosis mediated by the Fas antigen. It is hoped that this will be revealed.
  • the immune system elicits an immune response against foreign non-self antigens such as bacteria and viruses to eliminate non-self antigens and protect the body. However, it does not show an immune response to self-antigens and self-tolerance is established.
  • Such an immune response is established by antigen recognition by lymphocytic cells, mainly T cells, which are immunocompetent cells, and the nature of the immune tolerance of recognizing non-self antigens and not recognizing self antigens. Obtained mainly during the education of lymphoid stem cells in the thymus.
  • a T cell receptor having a reactivity above a certain threshold value against its own major histocompatibility antigen presented by thymic stromal cells. Only the lymphoid stem cells that have possessed undergo positive selection and proceed to the next differentiation stage (acquisition of MHC restriction).
  • lymphoid stem cells having reactivity (self-reactivity) above a certain threshold are excluded by negative selection called clone removal.
  • Lymphoid stem cells that have received negative selection and lymphocyte stem cells that have not received either selection result in cell death by apoptosis.
  • Mature T cells that are specifically expressed in peripheral tissues and migrated to the periphery without migrating to the thymus take a form of self-tolerance called clonal paralysis (Internal Medicine, Vol. 73, No. 4, No. 6 Pp. 637, 1994; Internal Medicine, Vol. 74, No. 3, pp. 508, 1994).
  • autoreactive T cells accumulate in the periphery.
  • B cells which are immunocompetent cells, and self-reactive B cells die by apoptosis.
  • autoreactive B cells accumulate in the periphery.
  • Such autoreactive T cells ⁇ self-reactive B cells, or autoantibodies against the T cells or B cells, may cause damage to their organisms and cause various diseases. Become. Diseases due to such self-reactive lymphocytes (abnormal self-tolerance) are at the center of so-called autoimmune diseases (Internal Medicine, Vol. 73, No. 4, pp. 635-639; History of Medicine) 170, No. 2, pp. 135-138, etc.).
  • RA rheumatoid arthritis
  • Fas ⁇ Fas antigen
  • autoimmune diseases As described above, the relationship between autoimmune diseases and apoptosis mediated by molecules such as the Fas antigen or the Fas ligand has not yet been elucidated, and also regulates apoptosis mediated by the Fas antigen. However, the possibility of treating autoimmune diseases has not been revealed at all.
  • Abnormal self-tolerance that is, associated with so-called autoimmune diseases and underlying diseases caused by abnormal turnover of self-reactive lymphocytes such as self-reactive T cells or self-reactive B cells and Z or biological constituent cells
  • Self-reactive lymphocytes such as self-reactive T cells or self-reactive B cells and Z or biological constituent cells
  • Secondary abnormal immune status and apoptosis And the relationship between molecules such as Fas antigens or Fas ligands and apoptosis may be useful in identifying such autoimmune diseases and secondary abnormal immune conditions for which there is still no effective treatment. It can be a great clue to the potential of treatment. Disclosure of the invention
  • the present inventors are thought to be deeply involved in the induction of apoptosis in order to elucidate the relationship between apoptosis and secondary abnormal immune status associated with such autoimmune diseases and underlying diseases.
  • the Fas antigen a monoclonal antibody that is highly reactive with the mouse Fas antigen and does not show lethal toxicity when administered to mice MRL / MpT—g 1 d / 1 d mouse
  • a model mouse that strongly presents antibodies not only with various autoimmune diseases including lymph node swelling but also with various collagen disease lesions
  • the present inventors have found that amelioration or treatment of an autoimmune disease can be achieved by administering an antibody reactive with the Fas antigen to a living body in need of treatment, and causing abnormalities in autoreactive lymphocytes and / or turnover.
  • the present invention discloses that it can be achieved by inducing apoptosis in unnecessary biological constituent cells that could not be eliminated from the living body by the above method.
  • the present invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising, as an active ingredient, an antibody reactive with a Fas antigen or a part of the antibody reactive with a Fas antigen, wherein the pharmaceutical composition comprises It is extremely useful for improving and treating immune diseases.
  • the pharmaceutical composition of the present invention is an antibody originally produced by the living body itself or a part thereof.
  • the toxicity to living organisms is extremely low as compared with a pharmaceutical composition comprising a compound obtained by chemical synthesis.
  • the constant region of the antibody may be removed to remove F (ab ') 2 or A part of an antibody such as a Fab, a chimeric antibody or a human-type antibody produced by gene recombination technology, or a human antibody produced by using a human antibody-producing transgenic nick mouse, etc. as an active ingredient By doing so, the immune rejection can be easily reduced.
  • the present invention is a pharmaceutical composition for treating an autoimmune disease, which comprises, as an active ingredient, an antibody reactive with a Fs antigen or a part of the antibody reactive with a Fs antigen.
  • the first of the specific aspects is a pharmaceutical composition for treating an autoimmune disease, comprising a monoclonal antibody reactive to a Fas antigen or a part thereof reactive to a Fas antigen as an active ingredient. .
  • the second aspect of the specific aspect is to treat an autoimmune disease comprising an antibody reactive with a membrane-bound Fas antigen or a part of the antibody reactive with a membrane-bound Fas antigen as an active ingredient.
  • a pharmaceutical composition comprising
  • the third aspect of the specific aspect is to treat an autoimmune disease comprising an antibody reactive with a secretory Fas antigen or a part of the antibody reactive with a secretory Fas antigen as an active ingredient.
  • an autoimmune disease comprising an antibody reactive with a secretory Fas antigen or a part of the antibody reactive with a secretory Fas antigen as an active ingredient.
  • the fourth aspect of the specific aspect is to treat an autoimmune disease, comprising, as an active ingredient, an antibody reactive with a human-derived Fas antigen or a part of the antibody reactive with a human-derived Fas antigen. And a pharmaceutical composition for
  • a fifth aspect of the specific aspect is an antibody reactive with a Fas antigen, which comprises an immunoglobulin variable region derived from a mammal other than human and a constant region of human-derived immunoglobulin, or a Fas antigen.
  • a pharmaceutical composition for treating an autoimmune disease which comprises a part of the antibody reactive with the antibody as an active ingredient.
  • the sixth aspect of the specific aspect is an antibody reactive with the Fas antigen, which comprises part or all of the hypervariable region of a non-human mammal-derived immunoglobulin, and a framework region of the human-derived immunoglobulin variable region. And a part of the antibody having reactivity with the Fas antigen as an active ingredient, which is a pharmaceutical composition for treating an autoimmune disease. .
  • a seventh aspect of the present invention is a pharmaceutical composition for treating an autoimmune disease, comprising as an active ingredient a human antibody reactive with a Fas antigen or a part of the antibody reactive with a Fas antigen. It is.
  • Eighth of the specific aspect is a pharmaceutical composition for treating an autoimmune disease, comprising F (ab ') 2 or Fab, which is a part of an antibody reactive with a Fas antigen, as an active ingredient. is there.
  • the “Fas antigen” in the present invention refers to a mammal, preferably a human, a mouse, a rat, a hamster, a guinea pig, a heron, a cat, a dog, a pig, a goat, a pig or a pig, and more Preferably, it is a Fas antigen (also referred to as CD95 or APO-1 antigen) derived from humans, cats, dogs, pigs, goats, pomas or pests, particularly preferably humans.
  • Fas antigen also referred to as CD95 or APO-1 antigen
  • Fas antigen CD95, APO-1 antigen
  • Cell Vol. 66, pp. 233-243, 1991
  • Journal of Experimental Medicine J. Exp. Med
  • Journal of Biological Chemistry J. Biol. Chem
  • Journal of Immunology Journal of I, unology
  • Fas antigen refers to a substance that penetrates or physically Derived from a membrane-bound Fas antigen that is scientifically bound and expressed on the cell surface, or a cDNA encoding the membrane-bound Fas antigen but different splicing proteins of the mRNA precursor Includes secreted (soluble) Fas antigens produced by degradation by proteolytic enzymes after expression.
  • the “antibody” in the present invention means a polyclonal antibody (antiserum) or a monoclonal antibody, and is preferably a monoclonal antibody.
  • the “antibody” of the present invention is a natural antibody obtained by immunizing an antigen (eg, a natural antigen, a recombinant antigen, a cell expressing an antigen, etc.) to a mammal as described above.
  • an antigen eg, a natural antigen, a recombinant antigen, a cell expressing an antigen, etc.
  • chimeric antibodies and humanized antibodies that can be produced using gene recombination techniques, and human antibodies that can also be produced using human antibody-producing transgenic animals and the like.
  • a monoclonal antibody having any isotype such as IgG, IgM, IgA, IgD or IgE is also included.
  • IgG IgG
  • IgM IgA
  • IgD IgD
  • IgE IgG
  • polyclonal antibody (antiserum) or monoclonal antibody referred to in the present invention can be produced by an existing general production method. That is, for example, antigen
  • a natural antigen, a recombinant antigen, a cell expressing the antigen, etc. can be used in mammals, preferably mice, rats, hamsters, guinea pigs, rabbits, cats, dogs, pigs, goats, dogs Alternatively, immunize a mouse, a rat, a hamster, a guinea pig, or a rabbit.
  • the polyclonal antibody can be obtained from serum obtained from the immunized animal.
  • a monoclonal antibody is prepared by preparing a hybridoma from an antibody-producing cell obtained from the immunized animal and a myeloma cell line (myeloma cell) having no autoantibody-producing ability, cloning the hybridoma, and immunizing a mammal.
  • a myeloma cell line myeloma cell having no autoantibody-producing ability
  • the monoclonal antibody can be specifically produced as follows. That is, a gene that expresses the Fas-expressing natural cell or the Fas antigen in various forms (membrane-bound, chimeric protein with other proteins or secreted (soluble), etc.) Using the engineered transformant itself or the various forms of the Fas antigen isolated and purified from those cells as an immunogen, the immunogen is used as a mouse, rat, hamster, guinea pig or rabbit, preferably mouse Subcutaneously, intramuscularly, intravenously, in rats or hamsters (including transgenic animals produced to produce antibodies from other animals, such as transgenic mice that produce human antibodies) Immunization is performed by one or several injections or transplantation into the footpad or intraperitoneal cavity. Usually, immunization is performed 1 to 4 times about every 1 to 14 days after the initial immunization, and antibody producing cells are obtained from the immunized mammal about 1 to 5 days after the final immunization.
  • the preparation of a hybridoma secreting a monoclonal antibody was carried out according to the method of Keraichi and Milshutine [Nature, Vol. 256, pp. 495-497, 1975] and It can be performed according to a modification method according to it.
  • myeloma cells used for cell fusion include mouse-derived myeloma P3 / X63-AG8.63 (653), P3 / NSI / 1-Ag4-1 (NS-1), P3 / X63-Ag8. P3U1), SP2 / 0-Agl4 (Sp20, Sp2), PAI, FO or BW5147, rat-derived myeloma 210RCY3-Ag. 2. 3., human-derived myeloma U-266AR1, GM1500- 6TG-Al-2, UC729-6, CEM-AGR, D1R11 or CEM-T15 can be used.
  • hybridoma clones producing monoclonal antibodies was performed by culturing hybridomas in, for example, a microtiter plate, and reacting the culture supernatant of the growing well with the immunizing antigen used in the mouse immunization described above. Can be measured by, for example, an enzyme immunoassay such as RIA or ELISA.
  • Production of the monoclonal antibody from the hybridoma is carried out in vitro or in vivo, for example, in a mouse, rat, guinea pig, hamster, or egret, preferably in a mouse or rat, more preferably in ascites of a mouse. It can be performed by isolation from the obtained culture supernatant or ascites of a mammal.
  • hybridomas When culturing in vitro, hybridomas are grown, maintained, and stored according to various conditions such as the characteristics of the cell type to be cultured, the purpose of the test and research, and the culture method, and the monoclonal antibody is produced in the culture supernatant. It can be carried out using any known nutrient medium or any nutrient medium derived and prepared from a known basal medium.
  • a Ham'F12 medium for example, a Ham'F12 medium, a low calcium medium such as an MCDB153 medium or a low calcium MEM medium, and a MCDB104 medium, a MEM medium, a D-MEM medium, High calcium media such as RPMI 164 medium, ASF 104 medium or RD medium can be used.
  • the basic medium may be, for example, serum, hormone, cytokine, and / or various inorganic or organic substances, depending on the purpose. Can be included.
  • an antibody containing a variable region of an immunoglobulin derived from a mammal other than a human and a constant region of an immunoglobulin derived from a human is genetically engineered.
  • variable region is a murine immunoglobulin-derived variable region and the constant region is a human immunoglobulin-derived constant region. It means a chimeric monoclonal antibody such as a mouse chimeric monoclonal antibody.
  • the constant region derived from human immunoglobulin has a unique amino acid sequence depending on its isotype, such as IgG, IgM, IgA, IgD and IgE.
  • the chimeric monoclonal antibody of the present invention May be the constant region of human immunoglobulin belonging to any of the isotypes, preferably the constant region of human IgG.
  • the chimeric monoclonal antibody in the present invention can be produced, for example, as follows. However, it is needless to say that the present invention is not limited to such a manufacturing method.
  • a mouse Z-human chimeric monoclonal antibody is produced with reference to Experimental Medicine (Extra Issue), Vol. 6, No. 10, 1988, and Japanese Patent Publication No. 3-732280. be able to. That is, under the active VH gene (rearranged VDJ gene encoding the H chain variable region) obtained from DNA encoding the mouse monoclonal antibody isolated from the hybridoma producing the mouse monoclonal antibody.
  • a CH gene C gene encoding the H chain constant region obtained from DNA encoding human immunoglobulin and an active gene obtained from DNA encoding a mouse monoclonal antibody isolated from the hybridoma were used.
  • CL gene C gene encoding L chain constant region
  • VL gene rearranged VJ gene encoding L chain variable region
  • DNA is extracted from a mouse monoclonal antibody-producing hybridoma by a conventional method, and the DNA is digested with an appropriate restriction enzyme (for example, EcoRI, HindIII, etc.), Perform the Southern blot method (for example, using 0.7% agarose gel).
  • the electrophoresed gel is stained with, for example, ethidium mouth, and after taking a photograph, the marker is attached, the gel is washed twice, and immersed in 0.25M HCl solution for 15 minutes. Then, immerse in 0.4N NaOH solution for 10 minutes while gently shaking. Transfer to the filter 1 by the usual method. After 4 hours, collect the filter and wash twice with 2XSSC. After the filter is completely dried, carry out paking (75 hours, 3 hours).
  • the filter After completion of the pacing, the filter is placed in a 0.1 lxSS CZO. 1% SDS solution and treated at 65 ° C for 30 minutes. Then soak in 3xSS C / 0.1% SDS solution. The obtained filter is put in a bininole bag together with the pre-hybridization solution, and treated with 65 pieces for 3 to 4 hours.
  • the probe PNA labeled with 32 P and the hybridization solution are added thereto, and the mixture is reacted at 65 ° C. for about 12 hours.
  • wash the filter under appropriate salt concentration, reaction temperature and time eg, 2XSSC-0.1 SDS solution, room temperature, 10 minutes. Put the filter in a bag of Vininole, add a small amount of 2XSSC, seal, and perform autoradiography.
  • the rearranged VDJ gene and VJ gene encoding the H chain and L chain of the mouse monoclonal antibody are identified by the Southern blot method described above.
  • the region containing the identified DNA fragment is fractionated by sucrose density gradient centrifugation, incorporated into a phage vector (for example, Charon 4A, Charon 28, ⁇ EMBL3, iEMBL4, etc.), and E. coli ( For example, LE392, M539, etc.) are transformed to produce a genomic library.
  • a phage vector for example, Charon 4A, Charon 28, ⁇ EMBL3, iEMBL4, etc.
  • E. coli For example, LE392, M539, etc.
  • plaque hybridization is performed to obtain positive clones each containing the rearranged VDJ gene or VJ gene.
  • a restriction enzyme map of the obtained clone is prepared, its nucleotide sequence is determined, and it is confirmed that the desired rearranged gene containing the rearranged VH (VDJ) gene or VL (VJ) gene has been obtained.
  • the human CH gene and human CL gene used for chimerization are isolated separately.
  • the Crl gene which is a CH gene and the C / c gene which is a CL gene are isolated.
  • These genes utilize the high homology of the nucleotide sequences of the mouse immunoglobulin gene and the human immunoglobulin gene to convert the mouse C1 gene and mouse C / c gene corresponding to the human C71 gene and human C / c gene. It can be obtained by using it as a probe and isolating it from a human genomic library.
  • clone 3 from Clone I gl 46 (Proc. Natl. Acad. Sci. USA), Vol. 75, pp. 4709-4713, 1978. 6.
  • Hindlll-kb BamHI fragment and clone MEP 10 (Pro Nat. Acad. Sci. USA, 78, 474-478, 1981).
  • a human lambda, Charon 4A from the Hael II-Alul Genomic Library-1 (Cell, Vol. 15, pp. 1157-1174, 1978) Isolate a DNA fragment containing the C / c gene and carrying the enhancer region.
  • human C 7 1 gene for example a human Bok fetal hepatocyte DNA is digested with Hindlll, after fractionated by Agarosugeru electrophoresis, a band of 5. 9 kb; inserted into I 788, using the above probe Isolate.
  • the human CH gene is placed downstream of the mouse VH gene while taking into account the promoter region and enhancer region.
  • mouse VL The human CL gene is inserted downstream of the gene into an expression vector such as pSV2gpt or pSV2neo using a suitable restriction enzyme and DNA ligase in a conventional manner.
  • the chimeric gene of the mouse VH gene Z human CH gene and the mouse VL gene noise CL gene may be simultaneously arranged in one expression vector, or may be arranged in separate expression vectors.
  • Chimeric gene-inserted expression vector was prepared, for example in T> such 3 ⁇ 63 ⁇ ⁇ ⁇ 8 ⁇ 653 cells or SP210 cells, protoplast fusion to myeloma cells that do not produce antibodies itself, DEAE-dextran method Introduced by the calcium phosphate method or the electroporation method.
  • the transformed cells are selected by culturing in a drug-containing medium corresponding to the drug resistance gene introduced into the expression vector, to obtain the desired chimeric antibody.
  • the desired chimeric monoclonal antibody is obtained from the culture supernatant of the antibody-producing cells thus selected.
  • an antibody comprising part or all of the hypervariable region of a non-human mammal-derived immunoglobulin, the framework region of the variable region of a human-derived immunoglobulin, and the constant region of a human-derived immunoglobulin is an antibody produced by genetic engineering, specifically, for example, a part or all of the complementarity determining region of the hypervariable region.
  • the framework region of the variable region is the framework region of the variable region derived from human immunoglobulin
  • the constant region is the human immunoglobulin derived region Means a human monoclonal antibody characterized by being a constant region.
  • the complementarity-determining regions of the hypervariable regions are three regions (Contplementarity-determining residue; CDRUCDR2, CDR3), and the framework region of the variable region is defined as four relatively conserved regions intervening before and after the three complementarity determining regions. Refers to the area (Framework; FR1, FR2, FR3, FR4).
  • it means a monoclonal antibody in which all regions other than part or all of the complementarity determining region of the hypervariable region of a mouse monoclonal antibody have replaced the corresponding region of human immunoglobulin.
  • the constant region derived from human immunoglobulin has an amino acid sequence unique to each of isotypes such as IgG, IgM, IgA, IgD and IgE, but the constant region of the humanized monoclonal antibody in the present invention It may be a constant region of human immunoglodaline belonging to the following isotype. Preferably, it is a human IgG constant region. Further, the framework region of the variable region derived from human immunoglobulin is not limited.
  • the humanized monoclonal antibody in the present invention can be produced, for example, as follows. However, it is needless to say that the present invention is not limited to such a manufacturing method.
  • a recombinant humanized monoclonal antibody derived from a mouse monoclonal antibody can be produced by genetic engineering with reference to Japanese Patent Application Laid-Open No. 4-506458 and Japanese Patent Application Laid-Open No. 62-296890. it can. That is, at least one mouse H chain CDR gene and at least one mouse L chain CDR gene corresponding to the mouse H chain CDR gene are isolated from a hybridoma producing a mouse monoclonal antibody.
  • a human H chain gene encoding the entire region other than the human H chain CDR corresponding to the mouse H chain CDR, and a human encoding the entire region other than the human L chain CDR corresponding to the pre-mouse L chain CDR Isolate the L chain gene.
  • the isolated mouse H chain CDR gene and the human H chain gene are introduced into an appropriate expression vector so that they can be expressed, and the mouse L chain CDR gene and the human L chain gene can be expressed similarly.
  • the mouse H chain CDR gene / human H chain gene and mouse L chain CDR gene The gene can also be introduced so that it can be expressed in the same expression vector.
  • human antibody means that all regions including the variable region of the H chain and the constant region of the H chain, and the variable region of the L chain and the constant region of the L chain which constitute immunoglobulin are human immunoglobulin. Is an immunoglobulin derived from the gene encoding
  • Human antibodies can be obtained by immunizing a transgenic animal produced by integrating at least the human immunoglobulin gene into a locus of a non-human mammal such as a mouse according to a conventional method, using an antigen. It can be produced in the same manner as in the method for producing a polyclonal antibody or a monoclonal antibody described above.
  • transgenic mice that produce human antibodies are described in Nature Genetics, Vol. 7, pp. 13-21, 1991; No. 5; International Application Publication WO94 / 255558; Nikkei Science, June, pp. 40-50, 1995, Nature, No. 36 8, pp. 856-858, pp. 1994; and Japanese Patent Application Laid-Open No. 6-500233, for example.
  • the “part of the antibody” in the present invention means the above-mentioned antibody in the present invention, preferably a partial region of the monoclonal antibody, and specifically, for example, “F (ab ′) 2”, “F ab” , [Fab] or "Fv”.
  • F (ab ′) 2 and “Fab ′” are produced by treating immoglobulin (monoclonal antibody) with pepsin or happine, which is a protease, in the hinge region. Means the antibody fragment produced by digestion before and after the disulfide bond existing between the two heavy chains.
  • the disulfide bond existing between the two heavy chains in the hinge region L chain consisting of VL (light chain variable region) binding (light chain constant region) and H consisting of VH (heavy chain variable region) and CH7I (71 region in heavy chain constant region)
  • Two homologous antibody fragments in which the chain fragments are linked by disulfide bonds in the C-terminal region can be produced.
  • Each of these two homologous antibody fragments is called Fab '.
  • IgG is treated with pepsin, it is cleaved downstream of the disulfide bond existing between the two heavy chains in the hinge region, and is slightly larger than that connected by the two Fab 'hinge regions. Can be manufactured. This antibody fragment is called F (ab ') 2 .
  • the antibody used in the present invention is an antibody having reactivity to the Fas antigen.
  • the Fas antigen is apoptosis of various cells. Is thought to be one of the molecules that mediate this.
  • monoclonal antibodies reactive to the Fas antigen by binding to the Fas antigen, reduce the fragmentation of DNA in the cell nucleus (DNA Fragmentation), one of the major features of the cell apoptosis phenomenon. It has been shown to cause cell death.
  • the antibody L used in the present invention is an antibody that induces apoptosis in cells by binding to a membrane-bound Fas antigen as described above. And a portion of the antibody.
  • the antibody or a part of the antibody used in the present invention includes an antibody reactive with a secretory Fas antigen as described above and a part of the antibody, and such an antibody or a part of the antibody is included.
  • a secretory Fas antigen as described above and a part of the antibody, and such an antibody or a part of the antibody is included.
  • autoimmune disease is used with the meaning as described below.
  • a living body has a function called an immune response (that is, a function that tries to remove non-self cells or antigens not derived from its own body) and a function called self tolerance (that is, a function derived from its own body).
  • an immune response that is, a function that tries to remove non-self cells or antigens not derived from its own body
  • self tolerance that is, a function derived from its own body.
  • Cells or antigens that do not have an immune response such as abnormalities in the clonal elimination mechanism in the thymus to establish immune tolerance and the clonal paralysis mechanism in peripheral tissues. If there is such an abnormality in the living body, a lymphocyte is generated in the living body that recognizes its own component as an antigen and elicits an immune response, despite being a lymphocyte derived from the living body.
  • Rinha sphere Clonal deletion in mosquito ⁇ the thymus which is a child of "phosphorus 80 ⁇ to elicit recognition by the immune response to any of the self-components for the antigen" generally "self-reactive Rinha sphere” in the present invention
  • the lymphocytes generated by abnormalities of the mechanism and the mechanism of clonal paralysis in peripheral tissues are naturally included in “self-reactive lymphocytes”.
  • “Rinno. Sphere” refers to T lymphocyte cells (T cells), B lymphocyte cells (B cells) and NK cells having various properties and functions at all stages of differentiation and maturation. (Natural killer cell).
  • the autoreactive lymphocytes, or antibodies (autoantibodies) produced by the immune response elicited by the autoreactive lymphocytes cause various lesions in the living body.
  • the “self-reactive lymphocyte” is defined as a disorder or disease caused by a cause unrelated to the abnormalities of the clonal elimination mechanism in the thymus and the clonal paralysis mechanism in the peripheral tissues (in the present invention, “ The term “underlying disease” may be used.
  • the pharmaceutical composition of the present invention is also used for improving or treating a secondary abnormal immune state caused by such autoreactive lymphocytes and the like.
  • autoimmune disease as used in the present invention is commonly used by those skilled in the art. As described above, it has the same meaning as “a disease in which the presence of autoreactive lymphocytes or autoantibodies is associated with the establishment of a lesion,” but in the present specification, at least the “autoreactive Diseases caused by “lymphocytes” are included in “autoimmune diseases”.
  • RA rheumatoid arthritis
  • the etiology is determined not only by the autoreactive lymphocytes and autoantibodies described above, but also by the disease site. It has been clarified that abnormal turnover of the synovial cells, which are the constituent cells of the living body, is part of the etiology.
  • the normal differentiation and proliferation of the constituent cells of the living body (neogenesis of cells), which are necessary to maintain the function of the living body normally, and the function of cells that have become old and have completed their given role
  • the balance of elimination, that is, cell turnover, is maintained.
  • RA rheumatoid arthritis
  • the normal elimination of old synovial cells at the site of disease is not performed, resulting in cell neogenesis.
  • the number of synovial cells will increase abnormally.
  • a pathological condition caused by "abnormal turnover of a constituent cell" is also included in the term "autoimmune disease”.
  • the pharmaceutical composition of the present invention relates to a disorder or disease that occurs as a primary or secondary cause of such abnormalities in the turnover of biological constituent cells, or the abnormalities in the tumors.
  • the secondary improvement of autoimmune diseases can be used for treatment.
  • the “autoimmune disease” in the present invention includes, specifically: (1) the following diseases caused by immune abnormality: autoimmune hemolytic anemia, idiopathic thrombocytopenic purpura, pernicious anemia, autoimmune thyroid Inflammation, myxedema, Graves' disease, autoimmune adrenalitis, idiopathic hypoparathyroidism, systemic sclerosis, systemic lupus erythematosus, rheumatoid arthritis, insulin-dependent diabetes mellitus, pageet's disease, membranous nephritis , Good Pastier's disease syndrome group, ulcerative colitis, atrophic gastritis, paralytic disease, myasthenia gravis, autoimmune hepatitis, self-sensitizing dermatitis, Siggren's syndrome, autoimmune encephalomyelitis, viral hepatitis 2 Abnormal immune symptoms associated with chronic respiratory disease 3 Abnormal immunity associated with subacute bacterial endocarditis ⁇ Post
  • rheumatoid arthritis systemic sclerosis, systemic lupus erythematosus, Siegren's syndrome, autoimmune hepatitis, myasthenia gravis, insulin-dependent diabetes mellitus, or viral hepatitis caused by immune disorders And other diseases.
  • the “pharmaceutical composition” in the present invention comprises, as an active ingredient, the antibody of the present invention or a part of the antibody, as a pharmaceutically acceptable carrier, that is, an excipient, a diluent, a bulking agent, a disintegrant, Pharmaceutical compositions with one or more of stabilizers, preservatives, buffers, emulsifiers, fragrances, coloring agents, sweeteners, thickeners, corrigents, solubilizers or other additives, tablets, pills It can be administered orally or parenterally in the form of powders, granules, injections, solutions, capsules, troches, elixirs, suspensions, emulsions or syrups.
  • 0.1 ag antibody 1 carrier to 1 mg antibody Zm in a non-toxic pharmaceutically acceptable carrier such as physiological saline, commercially available distilled water for injection or phosphate buffer. It can be produced by dissolving or suspending to a concentration of one carrier.
  • the injections produced in this way can be administered to human patients in need of treatment or livestock such as pests at a rate of 1 g / kg body weight / kg LOO mg per dose. It can be administered once to several times a day, preferably at a rate of 50 g to 50 mg.
  • the mode of administration can be a medically appropriate administration mode such as intravenous injection, subcutaneous injection, intradermal injection, intramuscular injection or intraperitoneal injection. Preferably, it is an intravenous injection.
  • the improvement and therapeutic effect of the so-called autoimmune disease and abnormal immunity state by the pharmaceutical composition of the present invention include the so-called autoimmune disease caused by autoreactive lymphocytes and the abnormal immunity state associated with the underlying disease.
  • Transgenic animals that are produced chemically and exhibit various abnormal immune symptoms (for example, transgenic mice into which a viral gene has been introduced).
  • C3H / Hej-g 1d / 1d mouse known as a model mouse for autoimmune disease caused by apoptotic signal transmission abnormality due to mutation of gene to be transfected [Cell, Vol. 76, No. 969-976, 1994 etc.], MRLZMpT—g 1 d / g 1 d mouse [Experimental Medicine, Vol. 13, No. 2, pp.
  • the antibody of the present invention contains a part of the antibody as an active ingredient and is pharmaceutically acceptable as described above.
  • a carrier that can be Various symptoms present in the model mouse or diseased animal, for example, autoimmune disease symptoms such as swelling of the lymph nodes, spleen enlargement, glomerulonephritis, salivary glanditis, pneumonia, arthritis, increased IgG or appearance of autoantibodies, Can be confirmed by observing the improvement of collagen disease symptoms.
  • FIG. 1 shows the hamster anti-mouse Fas monoclonal antibodies RK-8, SK-8, C6-1 and P4-4 (each indicated by a dotted line) and the control monoclonal antibody Hamster IgG (each indicated by a solid line).
  • FIG. 7 shows the reactivity of L5178Y cells not expressing the Fas antigen and L5178Y cells overexpressing the mouse Fas antigen. The vertical axis indicates the relative cell number, and the horizontal axis indicates the fluorescence intensity.
  • FIG. 2 shows the rat anti-mouse Fas monoclonal antibodies RMF-2, RMF-6, RMF-9 and RMF_13 (each indicated by a dotted line) and the control monoclonal antibody rat IgG (each indicated by a solid line).
  • FIG. 4 shows the reactivity of L5178Y cells not expressing the Fas antigen and L5178Y cells overexpressing the mouse Fas antigen. The vertical axis indicates the relative cell number, and the horizontal axis indicates the fluorescence intensity.
  • FIG. 3 shows hamster anti-mouse Fas monoclonal antibodies RK-8, SK-8, C6-1 and P4-4 (each indicated by a black waveform) and rat anti-mouse Fas monoclonal antibodies RMF-2, RMF- 6, B a 1 b / c mouse thymocytes of RM F-9 and RM F-13 (each indicated by a black waveform) and the control monoclonal antibody hamster IgG (each indicated by an open waveform)
  • FIG. 3 is a diagram showing the reactivity to The vertical axis indicates the relative cell number, and the horizontal axis indicates the fluorescence intensity.
  • FIG. 4 is a graph showing the effects of the hamster anti-mouse Fs monoclonal antibody RK-8 and the rat anti-mouse Fs monoclonal antibody RMF-2 on mouse Fs antigen overexpressing L5178Y cells.
  • FIG. 5 is a graph showing the effect of the hamster anti-mouse F a s monoclonal antibody RK-8 on Bal b / c mouse thymocytes and MRL + Z + mouse thymocytes.
  • FIG. 6 is a micrograph of a lymph node tissue section showing the state of lymph node swelling of a control 37-week-old mouse (left) and a 21-week-old RK-8-administered mouse at 37 weeks (right).
  • FIG. 7 is a microscopic photograph of a spleen tissue section showing the state of spleen swelling of a control 37-week-old mouse (left) and a 21-week-old RK-8-administered mouse at 37 weeks (right).
  • FIG. 8 is a microscopic photograph of a renal glomerular tissue section showing the state of glomerulonephritis in a control 5-week-old mouse.
  • FIG. 9 is a micrograph of a renal glomerular tissue section showing the state of glomerulonephritis in a control 21-week-old mouse.
  • FIG. 10 is a microscopic photograph of a glomerular tissue section of a 13-week-old RK-8-administered mouse showing the state of glomerulonephritis at the age of 21 weeks.
  • FIG. 11 is a microscopic photograph of a synovial tissue section showing the state of arthritis of a control 5-week-old mouse.
  • Figure 12 is a micrograph of a synovial tissue section showing the state of arthritis in control 21-week-old mice. It is.
  • FIG. 13 is a microscopic photograph of a synovial tissue section showing the state of arthritis of a 13-week-old RK-8-administered mouse at the age of 21 weeks.
  • FIG. 14 is a micrograph of a salivary gland tissue section showing the state of salivary glanditis in a control 5-week-old mouse.
  • FIG. 15 is a photomicrograph of a salivary gland tissue section showing the state of salivary glanditis in control 21-week-old mice.
  • FIG. 16 is a microscopic photograph of a salivary gland tissue section showing the state of salivary glanditis in a 13-week-old RK-8-administered mouse at the age of 21 weeks.
  • FIG. 17 is a photomicrograph of a lung tissue section showing the state of pneumonia in a control 5-week-old mouse.
  • FIG. 18 is a microscopic photograph of a lung tissue section showing the state of pneumonia in a control 21-week-old mouse.
  • FIG. 19 is a photomicrograph of a lung tissue section showing the state of pneumonia of a 13-week-old RK-8-administered mouse at the age of 21 weeks.
  • FIG. 2 is a cell distribution diagram showing the expression states of B220 antigen and T or T hy-1 antigen in each cell.
  • the numbers on the right shoulder of each distribution chart show the ratio (%) of the number of B220-positive ZThy-1 positive cells.
  • the immunizing antigen used to produce a monoclonal antibody reactive with the mouse Fas antigen was ligated to the extracellular region of the mouse Fas antigen using a genetic engineering technique. Soluble region) and mouse IL-3 receptor; S-subunit AIC2A extracellular region.
  • the E.co1i cell line JM109 transformed with an expression vector constructed to express this chimeric soluble mouse Fas antigen has been internationally deposited with the Ministry of International Trade and Industry's National Institute of Advanced Industrial Science and Technology. : FERM BP—4437, Contract date: October 6, 1993).
  • Plasmids expressing chimeric soluble mouse Fas antigen were extracted from the transformed cells JMl09 by a conventional method, and electoporation method (Nucl. Acids Res.), Vol. 16, Nos. 6127-6145 Page, 1988 etc.], the monkey-derived COS 7 cells were transformed with the plasmid.
  • Transformed C0S7 is obtained by culturing for 45 hours in D-MEM medium containing 10% fetal calf serum (FCS) and then culturing for 12 hours in serum-free D-MEM medium. The resulting serum-free culture was centrifuged and the centrifuged supernatant was collected.
  • FCS fetal calf serum
  • the obtained centrifuged supernatant was subjected to SDS-polyacrylamide gel electrophoresis to confirm that the desired chimeric soluble mouse Fas antigen was expressed, and the centrifuged supernatant was subjected to column chromatography using Q Sepharose. (Pharmacia, Inc.) to purify the chimeric soluble mouse Fas antigen.
  • a mixed solution of the obtained chimeric soluble mouse Fas antigen (100 / gZPBS) and killed bacteria (2 ⁇ 10 9 ) of Bordetella pertussis was administered intraperitoneally to Armenian hamsters. Furthermore, chimeric soluble mouse Fas antigen (10 g / PBS) was administered intraperitoneally. Three days after the administration, the hamster was laparotomized, the spleen was excised, triturated on a stainless steel mesh in a serum-free RPMI 1640 culture solution, and the spleen cell solution was centrifuged (1500 rpm for 7 minutes). The centrifugation residue was collected and suspended in serum-free RPMI 1640 medium. In addition, serum-free RPMI After washing twice with 1640 culture solution, antibody-producing hamster spleen cells were obtained.
  • each of the hybridomas was cultured in a serum-free ASF104 culture medium, and the culture supernatant was centrifuged (8000 rpm for 20 minutes). The centrifuged supernatant was collected, and each monoclonal antibody was purified by column chromatography using Protein A Sepharose (Pharmacia).
  • the rats were laparotomized and their spleens were excised, ground in a serum-free RPMI 1640 culture medium on a stainless steel mesh, and centrifuged (7 minutes at 1500 rpm). The centrifugation residue was collected and suspended in a serum-free RPMI 1640 culture medium. Furthermore, the cells were washed twice with a serum-free RPMI 1640 culture solution to obtain antibody-producing rat spleen cells.
  • Reactivity of culture supernatants of each hybridoma colony against Ba1bc mouse thymocytes expressing Fas antigen to screen hybridomas producing anti-mouse Fas monoclonal antibody. was measured by a flow cytometer, and four hybridoma clones producing anti-mouse Fas monoclonal antibodies, RMF2, RMF6, RMF9 and RMF13 were obtained.
  • Clone RMF6 has been deposited internationally with the Institute of Biotechnology, Ministry of International Trade and Industry of Japan (accession number: FERM BP-4394, accession date: August 25, 1993).
  • Each of the hybridomas was cultured in a serum-free ASF104 culture medium, and the culture supernatant was centrifuged (8000 rpm for 20 minutes). The centrifuged supernatant was collected and subjected to column chromatography using Protein G Sepharose (Pharmacia). Each monoclonal antibody was purified.
  • RGF2 was IgGl
  • RMF6 was IgG2a
  • RMF9 was IgG 2a
  • RMF13 was confirmed to be IgM.
  • the monoclonal antibodies RMF2 and RMF6 are commercially available from Institute for Medical Biology (Aichi).
  • the reactivity was caused by overexpression of mouse lymphoma cells L5178Y (ATCC CRL1722) not expressing the Fas antigen and mouse Fas antigen prepared by genetically introducing full-length mouse FascDNA. It was confirmed by comparing the reactivity with the L5178 Y cells.
  • mouse F ss antigen overexpressing L5178 Y cells were prepared as follows.
  • An expression vector (pMF1) obtained by introducing the full-length mouse Fasc DNA obtained by a conventional method into pBluescrip tKS (+) (Stratagene, Inc.) was digested with EcoRI, The cut ends were blunted using a blunt end kit (Takara Shuzo Co., Ltd.) to obtain a pMF1 fragment containing the full-length mouse Fas cDNA.
  • the expression vector pME18S (available from Dr. Kazuo Maruyama) was digested with XhoI, and the cut ends were blunt-ended using DNA end blunting kit (Takara Shuzo Co., Ltd.). A pME 18 S fragment was obtained.
  • This pME18S / mFas was digested with PvuI using pGene18S / mFas fragment by electroporation (290 V) using Gene Pulser (manufactured by Bio-Rad, Inc.).
  • a mouse lymphoma cell L5178Y (ATCC CRL1722) was co-transformed with a plasmid p MAMneo (manufactured by Clontech) fragment obtained by digestion with EcoRI.
  • the transformed cells were cultured in a medium containing the antibiotic G418 to select for G418-resistant clones and by limiting dilution to obtain L5178Y cells overexpressing mouse Fas antigen.
  • Mouse Linha that does not express the F s antigen Tumor cells L5178Y, the mouse Fas antigen over-expressing L5178Y cells, or the suspension of Th1 cells of Ba1bc mice, were treated with the anti-mouse Fas monoclonal antibodies obtained in Examples 2 and 3 ( PBS containing 20 g / m 1) was added, and the mixture was reacted on ice for 1 hour. Next, PBS containing FITC-labeled anti-rat IgG, FITC-labeled anti-hamster IgG or FITC-labeled anti-rat IgM (each 20; g / ml) was added and reacted for 1 hour. After washing each cell with PBS (4), the reactivity was analyzed by in situ fluorimetry.
  • anti-rat IgG anti-hamster IgG and anti-rat IgM (Pharmingen (manufactured by Pharmingen) were used as controls for the same measurement.
  • Each of the obtained monoclonal antibodies was strongly confirmed to be a monoclonal antibody having specific reactivity to mouse Fas antigen.
  • mice Using a 96-well microplate, (1) mouse Fas antigen overexpressing L5178 Y cell prepared in Example 4 in RPMI 1640 medium, (2) Ba1b / c mouse (Charles River )) Thymocytes, and (3) MRL + Z + mice (Charles River (manufactured by)) Each of the thymocytes was cultured at a cell number of 1 ⁇ 10 5 / ⁇ . To each well, hamster anti-mouse Fas monoclonal antibody RK-8 or rat anti-mouse Fas monoclonal antibody RMF-2, diluted to various concentrations with RPMI 1640 culture medium containing 10% FCS, was added and incubated for 20 hours. .
  • Example 6 Therapeutic effect of autoimmune disease in animal model
  • MRLZMpT—gl dZgld mice were kindly provided by Dr. Masato Nose of Tohoku University School of Medicine.
  • MRLZMpT—g1 ⁇ / g1d mice are expressed in MRL / Mp— + Z + mice (normal mice) and in a mutation in the gene encoding the Fas ligand.
  • a mouse obtained by crossing He j -g 1 d / g 1 d mouse, in which expression of normal Fas ligand is suppressed as in C 3H / H ej -g 1 d / g 1 d mouse Can cause severe autoimmune disease symptoms (Experimental Medicine, Vol. 13, No. 2, pp.
  • Characteristic symptoms are lymph node swelling due to accumulation of abnormal lymphocytes (B220 positive ZT hy-1 positive g1d cells), spleen enlargement, severe glomerulitis, salivary glanditis, pneumonia , Arthritis, and the appearance of autoantibodies.
  • 13-week-old MRL / MpT-gldZgld mice (3 mice, hereinafter referred to as "13-week-old RK-8-administered mice") that began to exhibit symptoms of autoimmune diseases such as swelling of lymph nodes.
  • each of the 21-week-old MRLZMp T—g 1 d /] d mice (3 mice, hereinafter referred to as “21-week-old RK-8-administered mice”) in which the symptom progressed were treated with phosphoric acid.
  • the anti-mouse Fas monoclonal antibody RK-8 prepared in Example 2 dissolved in a buffer and used as an injection was administered intraperitoneally at a concentration of 200 mice.
  • control mice were similarly stained and used for tissue sections. The results are shown in FIGS.
  • the monoclonal antibodies for measurement include FITC (fluorescein isothiosinate) -labeled anti-mouse B220 monoclonal antibody (Pharmingen (manufactured by)) and PE (picoerythrin) -labeled anti-mouse Thy-1 monoclonal antibody (Pharmingen (manufactured by Pharmingen). )) was used.
  • FITC fluorescein isothiosinate
  • PE picoerythrin
  • spleen, lymph node and thymus-derived cells obtained in the same manner from normal MRLZMp-++ mice and control 21-week-old mice were used. The results are shown in FIG.
  • the upper right area shows the abnormal lymphocyte B220-positive and Thy-1 positive g1d cell groups and the right shoulder of the area. Numerical values indicate the percentage of the number of cells in the abnormal lymphocyte.
  • abnormal spontaneous lymphocyte B220-positive and Thy-1-positive g1d cells
  • Anti-ss DNA antibody collected from 13-week-old RK-8-administered mice (laparotomy 8 weeks after antibody administration) and 21-week-old RK-8-administered mice (laparotomy 4 weeks after antibody administration)
  • the antibody titers of ss: single strand (single strand)) and anti-ds DNA antibody (ds: double strand (double strand)) were confirmed by measuring the absorbance at a wavelength of 405 nm.
  • sera obtained in the same manner from control 21-week-old mice and control 25-week-old mice were used.
  • Serum from 11-week-old MRLZMp ⁇ + ⁇ + mice was used as a negative control. Table 1 shows the results. table 1
  • Table 2 schematically shows the results obtained in the above (1) to (3) and other results.
  • the score was + when the growth of mesangial cells and vitreous droplets were observed.
  • the present invention achieves, for the first time, improvement and treatment of an autoimmune disease by using an antibody reactive with the Fs antigen or a part of the antibody reactive with the Fs antigen.
  • the present invention comprises, as an active ingredient, an antibody reactive with a Fas antigen or a part of an antibody reactive with a Fas antigen, and is extremely useful for improving and treating autoimmune diseases. It is the first to provide a pharmaceutical composition.
  • the pharmaceutical composition of the present invention since the pharmaceutical composition of the present invention originally uses an antibody produced by the living body as an active ingredient, it has extremely low toxicity to living organisms as compared with a pharmaceutical composition composed of a chemically synthesized compound, When a mammal-derived antibody is applied to a patient as an active ingredient, F (ab ') 2 or F ab etc., which is obtained by removing the constant region of the antibody, even if there is a concern about immune rejection
  • the immunological rejection reaction can be carried out by using a part of the antibody, a chimeric antibody or a humanized antibody produced by gene recombination technology, or a human antibody produced using a human antibody-producing transgenic mouse as an active ingredient. It can be easily reduced.

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Abstract

Cette invention concerne une composition médicinale permettant de traiter des maladies auto-immunes, laquelle composition contient en qualité d'ingrédient actif, un anticorps capable de réagir à un antigène Fas ou à une partie de celui-ci.
PCT/JP1995/001778 1995-09-07 1995-09-07 Remede contre maladies auto-immunes WO1997009065A1 (fr)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP7181676B2 (ja) 2015-08-10 2022-12-01 東レ株式会社 免疫誘導剤

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
CHEMICAL ABSTRACTS, Vol. 120, (1994), Abstract No. 74852, ADACHI, M. et al.; & NIPPON NAIKA GAKKAI ZASSHI, 1993, 82, (10), 1732-7. *
J. BIOCHEM. Vol. 117, No. 5, (1995), p936-939, (KIKUCHI H. et al.). *
JOURNAL OF CLINICAL AND EXPERIMENTAL MEDICINE, (IGAKU NO AYUMI), Vol. 170, No. 2, (1994), p. 131-138, (KOBAYASHI S. et al.). *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP7181676B2 (ja) 2015-08-10 2022-12-01 東レ株式会社 免疫誘導剤

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