Therapeutic agent for autoimmune diseases or allergy, and method for screening for the therapeutic agent

Abstract

Disclosed is a therapeutic agent for treating a cellular immune disease, comprising as an active ingredient a substance that inhibits binding between Sema3A and a Neuropilin-1/Plexin-A1 heteroreceptor. The substance includes, for example, a Sema3A neutralizing antibody, a Neuropilin-1 neutralizing antibody, or a soluble Neuropilin-1 or derivative thereof. Also disclosed is a method for screening a therapeutic agent for treating a cellular immune disease utilizing a signal generated by the interactions of Neuropilin-1, Plexin-A1 and Sema3A as a marker.

1 . A therapeutic agent comprising as an active ingredient a substance that inhibits binding between a Neuropilin-1/Plexin-A1 heteroreceptor and Sema3A, wherein the therapeutic agent is suitable for treating a cellular immune disease. 
     
     
         2 . The therapeutic agent of  claim 1 , wherein the substance is a Sema3A neutralizing antibody. 
     
     
         3 . The therapeutic agent of  claim 2 , wherein the Sema3A neutralizing antibody binds to a peptide having a sequence of residue Nos. 363 to 381 of SEQ ID NO: 1. 
     
     
         4 . The therapeutic agent of  claim 3 , wherein the peptide has a sequence of NYQWVPYQGRVPYPRPGTC (SEQ ID NO: 12). 
     
     
         5 . The therapeutic agent of  claim 1 , wherein the substance is a Neuropilin-1 neutralizing antibody. 
     
     
         6 . The therapeutic agent of  claim 5 , wherein the Neuropilin-1 neutralizing antibody binds to a peptide having a sequence of residue Nos. 265 to 857 of SEQ ID NO: 2. 
     
     
         7 . The therapeutic agent of  claim 1 , wherein the substance is a neutralizing antibody that is a polyclonal antibody. 
     
     
         8 . The therapeutic agent of  claim 1 , wherein the substance is a neutralizing antibody that is a monoclonal antibody. 
     
     
         9 . The therapeutic agent of  claim 1 , wherein the substance comprises a soluble Neuropilin-1. 
     
     
         10 . The therapeutic agent of  claim 9 , wherein the soluble Neuropilin-1 is a polypeptide selected from the group consisting of:
 (1) a polypeptide having a sequence of residue Nos. 23 to 589 of SEQ ID NO: 2,   (2) a polypeptide having a sequence of residue Nos. 23 to 857 of SEQ ID NO: 2,   (3) a polypeptide that has a sequence comprising one or more amino acid deletions, additions or substitutions in the sequence of the polypeptide of (1) or (2), while retaining a Plexin-A1-Sema3A binding activity,   (4) a polypeptide encoded by a first polynucleotide that hybridizes under high stringent conditions with a second polynucleotide that encodes the polypeptide of (1) or (2), and which has a Plexin-A1-Sema3A binding activity, and   (5) a polypeptide encoded by a third polynucleotide that hybridizes under high stringent conditions with a fourth polynucleotide having a sequence of SEQ ID NO: 5, and which has a Plexin-A1-Sema3A binding activity.   
     
     
         11 . The therapeutic agent of  claim 1 , wherein the substance is a soluble Neuropilin-1 derivative. 
     
     
         12 . The therapeutic agent of  claim 10 , wherein the substance is a soluble Neuropilin-1 derivative that is a fused polypeptide of Fc and the soluble Neuropilin-1. 
     
     
         13 . The therapeutic agent of  claim 12 , wherein the Fc comprises a polypeptide having a sequence selected from the group consisting of SEQ ID NOS: 7, 8, 9 and 10. 
     
     
         14 . The therapeutic agent of  claim 10 , wherein the substance is a soluble Neuropilin-1 derivative that is a polypeptide comprising a polyethylene glycol chain added to the soluble Neuropilin 1. 
     
     
         15 . The therapeutic agent of  claim 1 , wherein the cellular immune disease is an autoimmune disease. 
     
     
         16 . The therapeutic agent of  claim 15 , wherein the autoimmune disease is an organ-specific autoimmune disease. 
     
     
         17 . The therapeutic agent of  claim 16 , wherein the organ-specific autoimmune disease is selected from the group consisting of aplastic anemia, hemolytic anemia, autoimmune hemolytic anemia, idiopathic thrombocytopenia, autoimmune hepatitis, iridocyclitis, scleritis, uveitis, orchitis, idiopathic thrombocytopenia purpura, Basedow's disease, Hashimoto's thyroiditis, juvenile-onset diabetes, inflammatory bowel disease, Addison's disease, demyelinating encephalitis and multiple sclerosis. 
     
     
         18 . The therapeutic agent of  claim 15 , wherein the autoimmune disease is a systemic autoimmune disease. 
     
     
         19 . The therapeutic agent of  claim 18 , wherein the systemic autoimmune disease is atopic dermatitis, chronic rheumatoid arthritis or other arthritis, systemic lupus erythematosus, Sjogren's syndrome, undifferentiated connective tissue disease, antiphospholipid syndrome, a form of vasculitis, Wegener's granulomatosis, Kawasaki disease, hypersensitive angitis, Henoch-Schonlein purpurea, Behcet's disease, Takayasu's arteritis, giant cell arteritis, thromboangitis obliterans, polymyalgia rheumatica, essential (mixed) cryoglobulinemia, psoriasis, psoriasis vulgaris and psoriatic arthritis, diffuse fascitis with or without eosinophilia, recurrent panniculitis, recurrent polychondritis, lymphomatoid granulomatosis, erythema nodosum, ankylosing spondylitis, Reiter's syndrome, or a form of inflammatory dermatitis. 
     
     
         20 . The therapeutic agent of  claim 1 , wherein the cellular immune disease is a delayed-type allergic disease. 
     
     
         21 . The therapeutic agent of  claim 20 , wherein the delayed-type allergic disease is contact dermatitis, metal dermatitis, allergic contact dermatitis, Sjogren's syndrome, infectious allergy, drug-induced pneumonia or Guillan-Barre syndrome. 
     
     
         22 . A method for screening a therapeutic agent for treating a cellular immune disease, comprising:
 (a) contacting a Sema3A polypeptide with a polypeptide having a Neuropilin-1 extracellular domain, in the presence of a test substance;   (b) measuring a signal produced by an interaction between the Sema3A polypeptide and the polypeptide having a Neuropilin-1 extracellular domain in the presence of the test substance, and comparing the signal with a control signal produced by an interaction between the Sema3A polypeptide and the polypeptide having a Neuropilin-1 extracellular domain in the absence of the test substance; and   (c) selecting a test substance that reduces the signal in comparison to the control signal.   
     
     
         23 . A method for screening a therapeutic agent for treating a cellular immune disease, comprising:
 (a) contacting a Sema3A polypeptide with eukaryotic cells expressing Neuropilin-1 and Plexin-A1, in the presence of a test substance;   (b) measuring a signal produced by an interaction between the Sema3A polypeptide and the eukaryotic cells expressing Neuropilin-1 and Plexin-A1 in the presence of the test substance, and comparing the signal with a control signal produced by an interaction between the Sema3A polypeptide and the eukaryotic cells expressing Neuropilin-1 and Plexin-A1 in the absence of the test substance; and   (c) selecting a test substance that reduces the signal in comparison to the control signal.   
     
     
         24 . The method of  claim 23 , wherein the eukaryotic cells are dendritic cells. 
     
     
         25 . The method of  claim 23 , wherein the dendritic cells are cells fractionated and induced from peripheral blood. 
     
     
         26 . The method of  claim 23 , wherein the dendritic cells are cells of a cell line. 
     
     
         27 . The method of  claim 23 , wherein the signal is Rho kinase activation. 
     
     
         28 . The method of  claim 23 , wherein the signal is myosin II phosphorylation. 
     
     
         29 . The method of  claim 23 , wherein the signal is actomyosin contraction. 
     
     
         30 . The method of  claim 24 , wherein the signal is transmigration of the dendritic cells. 
     
     
         31 . A method for inhibiting activation of Rho kinase in dendritic cells expressing Neuropilin-1 and Plexin-A1, the method comprising inhibiting binding between Sema3A and Neuropilin-1 expressed on a surface of human dendritic cells. 
     
     
         32 . A method for inhibiting phosphorylation of myosin-II in dendritic cells expressing Neuropilin-1 and Plexin-A1, the method comprising inhibiting binding between Sema3A and Neuropilin-1 expressed on a surface of human dendritic cells. 
     
     
         33 . A method for inhibiting actomyosin contraction in dendritic cells expressing Neuropilin-1 and Plexin-A1, the method comprising inhibiting binding between Sema3A and Neuropilin-1 expressed on a surface of human dendritic cells.