Neutrokine-alpha antibodies and methods of use thereof

Abstract

The present invention relates to a Neutrokine-alpha antibody and a process for producing a Neutrokine-alpha antibody. The invention further relates to screening methods for identifying compounds that inhibit or enhance the action of Neutrokine-alpha. Also provided are diagnostic methods for detecting autoimmune disorders and therapeutic methods for treating autoimmune disorders using a Neutrokine-alpha antibody.

1 . A method of evaluating total Neutrokine-alpha in a biological sample, the method comprising:
 (a) contacting a biological sample with an anti-Neutrokine-alpha antibody that binds Neutrokine-alpha independent of its physiological state; and   (b) evaluating the amount of Neutrokine-alpha bound by the antibody,   wherein the amount of Neutrokine-alpha determined in (b) indicates total Neutrokine-alpha in the biological sample.   
     
     
         2 . A method of evaluating the distribution of Neutrokine-alpha in a biological sample, the method comprising:
 (a) contacting a biological sample with a first anti-Neutrokine-alpha antibody, which binds Neutrokine-alpha independent of its physiological state;   (b) contacting the sample with a second anti-Neutrokine-alpha antibody, which preferentially binds free Neutrokine-alpha over Neutrokine-alpha bound to receptor; and   (c) comparing the binding of the first antibody and the binding of the second antibody in the sample,   wherein the comparison in (c) indicates the distribution of Neutrokine-alpha among various physiological pools in the sample.   
     
     
         3 . A method of evaluating the distribution of Neutrokine-alpha, the method comprising:
 (a) contacting a first biological sample with a first anti-Neutrokine-alpha antibody, which binds Neutrokine-alpha independent of whether Neutrokine-alpha is bound to a receptor;   (b) contacting a second biological sample, of the same type as the first, with a second anti-Neutrokine-alpha antibody, which preferentially binds free Neutrokine-alpha over Neutrokine-alpha bound to receptor; and   (c) comparing the binding of the first antibody and the binding of the second antibody in the two samples,   wherein the comparison in (c) indicates the distribution of Neutrokine-alpha among various physiological pools in the samples.   
     
     
         4 . A method of evaluating a compound or composition for the ability to modulate total Neutrokine-alpha, the method comprising:
 (a) administering a test compound or composition to a mammal;   (b) obtaining a biological sample from the mammal;   (c) contacting the sample with an anti-Neutrokine-alpha antibody that binds Neutrokine-alpha independent of whether Neutrokine-alpha is bound to a receptor;   (d) evaluating the amount of Neutrokine-alpha bound by the antibody in the biological sample; and   (e) comparing the amount of Neutrokine-alpha bound by the antibody in the biological sample with the amount of Neutrokine-alpha bound by the antibody in a control sample,   wherein a difference in binding attributable to the test compound or composition indicates that the compound or composition is effective in modulating total Neutrokine-alpha in the mammal.   
     
     
         5 . A method of evaluating a compound or composition for the ability to modulate the distribution of Neutrokine-alpha, the method comprising:
 (a) administering a test compound or composition to a mammal;   (b) obtaining a biological sample from the mammal;   (c) contacting the sample with a first anti-Neutrokine-alpha antibody, which binds Neutrokine-alpha independent of its physiological state;   (d) contacting the sample with a second anti-Neutrokine-alpha antibody, which preferentially binds free Neutrokine-alpha over Neutrokine-alpha bound to Neutrokine-alpha receptor;   (e) detecting binding of the first antibody and the second antibody to Neutrokine-alpha in the sample; and   (f) comparing the binding of the first antibody and the second antibody in the biological sample to the binding of the first and second antibodies in a control sample,   wherein a difference in binding attributable to the test compound indicates that the compound or composition is effective in modulating the distribution of Neutrokine-alpha in the mammal.   
     
     
         6 . A method of evaluating a test compound or composition for the ability to modulate the distribution of Neutrokine-alpha, comprising:
 (a) administering a test compound or composition to a mammal;   (b) obtaining a first biological sample and a second biological sample, both of the same type, from the mammal;   (c) contacting the first biological sample with a first anti-Neutrokine-alpha antibody, which binds Neutrokine-alpha independent of whether Neutrokine-alpha is bound to a receptor;   (d) contacting the second biological sample with a second anti-Neutrokine-alpha antibody, which preferentially binds free Neutrokine-alpha over Neutrokine-alpha bound to receptor;   (e) detecting binding of the first antibody and the second antibody to Neutrokine-alpha in the two biological samples; and   (f) comparing the binding of the first antibody and second antibody in the two biological samples to the binding of the first and second antibodies, respectively, in one or more control samples,   wherein a difference in binding attributable to the test compound or composition indicates that the compound or composition is effective in modulating the distribution of Neutrokine-alpha in the mammal.   
     
     
         7 . The method of  claim 1 , wherein the biological sample or samples is or are obtained from a mammal that has an autoimmune disease. 
     
     
         8 . The method of  claim 1 , wherein the binding is evaluated qualitatively. 
     
     
         9 . The method of  claim 1 , wherein the binding is evaluated quantitatively. 
     
     
         10 . The method of  claim 1 , wherein the biological sample or samples comprises or comprise serum and the method comprises ELISA. 
     
     
         11 . The method of  claim 1 , wherein the biological sample or samples comprises or comprise blood and the method comprises flow cytometry. 
     
     
         12 . The method of  claim 1 , wherein the biological sample or samples comprises or comprise tissue and the method comprises immunohistochemistry. 
     
     
         13 . The method of  claim 1 , wherein the biological sample or samples comprises or comprise serum, blood, plasma, biopsy sample, tissue sample, cell suspension, biological fluid, saliva, oral fluid, cerebrospinal fluid, amniotic fluid, milk, colostrum, mammary gland secretion, lymph, urine, sweat, lacrimal fluid, gastric fluid, synovial fluid, or mucus. 
     
     
         14 . The method of  claim 2 , wherein the binding of the first anti-Neutrokine-alpha antibody indicates total Neutrokine-alpha. 
     
     
         15 . The method of  claim 2 , wherein the binding of the second anti-Neutrokine-alpha antibody indicates free Neutrokine-alpha. 
     
     
         16 . The method of  claim 2 , wherein the difference in binding of the first and second antibodies indicates receptor-bound Neutrokine-alpha in the sample. 
     
     
         17 . The method of  claim 4 , wherein the control sample is obtained from the same mammal prior to administration of the compound or composition to be tested. 
     
     
         18 . The method of  claim 4 , wherein the control sample is obtained from the same mammal prior to the onset of autoimmune disease. 
     
     
         19 . The method of  claim 4 , wherein the control sample is obtained from a healthy mammal. 
     
     
         20 . An isolated anti-Neutrokine-alpha antibody selected from the group consisting of:
 (a) an antibody that binds Neutrokine-alpha independent of its physiological state; and   (b) an antibody that binds to an epitope of Neutrokine-alpha outside of the binding sites of BCMA, TACI, and BR3.   
     
     
         21 . The antibody of  claim 20 , wherein the affinity of the antibody for free Neutrokine-alpha and the affinity of the antibody for Neutrokine-alpha bound to Neutrokine-alpha receptor differ by less than 30%. 
     
     
         22 . The antibody of  claim 20 , wherein the antibody binds to Neutrokine-alpha bound to Neutrokine-alpha receptor with an affinity of at least 10 6  M −1 . 
     
     
         23 . A method of selecting the antibody of  claim 20 , comprising determining whether the antibody binds Neutrokine-alpha independent of whether Neutrokine-alpha is bound to a receptor. 
     
     
         24 . A method of using the antibody of  claim 20 , comprising contacting a biological sample with the antibody, thereby evaluating the biological sample. 
     
     
         25 . A method of identifying a subject who is a candidate for a Neutrokine-alpha antagonist therapy, the method comprising:
 (a) contacting a biological sample from the subject with an anti-Neutrokine-alpha antibody that binds Neutrokine-alpha independent of its physiological state;   (b) evaluating the level of total Neutrokine-alpha bound by the antibody; and   (c) selecting a subject who has a greater level of total Neutrokine-alpha bound by the antibody, compared to a control, wherein a greater level of total Neutrokine-alpha bound by the antibody, compared to a control, indicates that the subject is a candidate for a Neutrokine-alpha antagonist therapy.   
     
     
         26 . The method of  claim 25 , wherein the Neutrokine-alpha antagonist therapy is selected from the group consisting of: an inhibitory anti-Neutrokine-alpha antibody and an anti-Neutrokine-alpha receptor antibody. 
     
     
         27 . The method of  claim 25 , wherein the subject has an autoimmune disease selected from the group consisting of: autoimmune rheumatologic disorders, autoimmune gastrointestinal and liver disorders, vasculitis, autoimmune neurological disorders, autoimmune dermatologic disorders, autoimmune endocrine disorders, renal disorders, and hematologic disorders. 
     
     
         28 . The method of  claim 25 , wherein the biological sample is selected from the group consisting of: blood, tissue, plasma, synovial fluid, and cerebrospinal fluid. 
     
     
         29 . The method of  claim 25 , wherein the subject has a normal level of free soluble Neutrokine-alpha compared to the control. 
     
     
         30 . A method of identifying a subject who is a candidate for a Neutrokine-alpha antagonist therapy, the method comprising:
 (a) contacting a biological sample from the subject with a first anti-Neutrokine-alpha antibody, which binds Neutrokine-alpha independent of its physiological state;   (b) contacting the biological sample with a second anti-Neutrokine-alpha antibody, which preferentially binds free Neutrokine-alpha over Neutrokine-alpha bound to receptor; and   (c) comparing the binding of the first antibody and the second antibody, thereby evaluating the level of receptor-bound Neutrokine-alpha,   wherein a greater level of receptor-bound Neutrokine-alpha, compared to a control, indicates that the subject is a candidate for a Neutrokine-alpha antagonist therapy.   
     
     
         31 . The method of  claim 30 , wherein the Neutrokine-alpha antagonist therapy is selected from the group consisting of: an inhibitory anti-Neutrokine-alpha antibody and an anti-Neutrokine-alpha receptor antibody. 
     
     
         32 . The method of  claim 30 , wherein the subject has an autoimmune disease selected from the group consisting of: autoimmune rheumatologic disorders, autoimmune gastrointestinal and liver disorders, vasculitis, autoimmune neurological disorders, autoimmune dermatologic disorders, autoimmune endocrine disorders, renal disorders, and hematologic disorders. 
     
     
         33 . The method of  claim 30 , wherein the biological sample is selected from the group consisting of: blood, tissue, plasma, synovial fluid, and cerebrospinal fluid. 
     
     
         34 . The method of  claim 30 , wherein the subject has a normal level of free Neutrokine-alpha compared to the control. 
     
     
         35 . A method of identifying a subject who is a candidate for a Neutrokine-alpha antagonist therapy, the method comprising:
 (a) contacting a first biological sample from the subject with a first anti-Neutrokine-alpha antibody, which binds Neutrokine-alpha independent of whether Neutrokine-alpha is bound to a receptor;   (b) contacting a second biological sample from the subject, of the same type as the first, with a second anti-Neutrokine-alpha antibody, which preferentially binds free Neutrokine-alpha over Neutrokine-alpha bound to receptor; and   (c) comparing the binding of the first antibody and the binding of the second antibody, thereby evaluating the level of receptor-bound Neutrokine-alpha,   wherein a greater level of receptor-bound Neutrokine-alpha, compared to a control, indicates that the subject is a candidate for a Neutrokine-alpha antagonist therapy.   
     
     
         36 . The method of  claim 35 , wherein the Neutrokine-alpha antagonist therapy is selected from the group consisting of: an inhibitory anti-Neutrokine-alpha antibody and an anti-Neutrokine-alpha receptor antibody. 
     
     
         37 . The method of  claim 35 , wherein the subject has an autoimmune disease selected from the group consisting of: autoimmune rheumatologic disorders, autoimmune gastrointestinal and liver disorders, vasculitis, autoimmune neurological disorders, autoimmune dermatologic disorders, autoimmune endocrine disorders, renal disorders, and hematologic disorders. 
     
     
         38 . The method of  claim 35 , wherein the biological sample type is selected from the group consisting of: blood, tissue, plasma, synovial fluid, and cerebrospinal fluid. 
     
     
         39 . The method of  claim 35 , wherein the subject has a normal level of free Neutrokine-alpha compared to the control. 
     
     
         40 . A method of monitoring a patient who has been administered a therapy for a B cell mediated disease, the method comprising performing the method of  claim 1  on a biological sample from the patient, wherein a decrease in total Neutrokine-alpha in the biological sample indicates that the patient is responsive to the therapy. 
     
     
         41 . The method of  claim 40 , wherein the method is performed at least 2 weeks after the administration of the therapy. 
     
     
         42 . The method of  claim 40 , wherein the method is performed at least twice at least 2 weeks apart after the administration of the therapy. 
     
     
         43 . The method of  claim 40 , wherein the subject has an autoimmune disease selected from the group consisting of: autoimmune rheumatologic disorders, autoimmune gastrointestinal and liver disorders, vasculitis, autoimmune neurological disorders, autoimmune dermatologic disorders, autoimmune endocrine disorders, renal disorders, and hematologic disorders. 
     
     
         44 . The method of  claim 40 , wherein the biological sample is selected from the group consisting of: blood, tissue, plasma, synovial fluid, and cerebrospinal fluid. 
     
     
         45 . The method of  claim 41 , wherein the subject has a normal level of free Neutrokine-alpha compared to the control. 
     
     
         46 . A method of evaluating total Neutrokine-alpha in a biological sample, the method comprising:
 (a) contacting a biological sample with an anti-Neutrokine-alpha antibody that binds Neutrokine-alpha ligand and Neutrokine-alpha ligand-receptor complexes, whether the Neutrokine-alpha is soluble or membrane-bound; and   (b) evaluating the amount of Neutrokine-alpha bound by the antibody,   wherein the amount of Neutrokine-alpha determined in (b) indicates total Neutrokine-alpha in the biological sample.   
     
     
         47 . A method of evaluating the distribution of Neutrokine-alpha in a biological sample, the method comprising:
 (a) contacting a biological sample with a first anti-Neutrokine-alpha antibody, which binds Neutrokine-alpha ligand and Neutrokine-alpha ligand-receptor complexes, whether the Neutrokine-alpha is soluble or membrane-bound;   (b) contacting the sample with a second anti-Neutrokine-alpha antibody, which binds to soluble Neutrokine-alpha; and   (c) comparing the binding of the first antibody and the binding of the second antibody in the sample,   wherein the comparison in (c) indicates the distribution of Neutrokine-alpha among various physiological pools in the sample.   
     
     
         48 . A method of evaluating a compound or composition for the ability to modulate the distribution of Neutrokine-alpha, the method comprising:
 (a) administering a test compound or composition to a mammal;   (b) obtaining a biological sample from the mammal;   (c) contacting the sample with a first anti-Neutrokine-alpha antibody, which binds Neutrokine-alpha ligand and Neutrokine-alpha ligand-receptor complexes, whether the Neutrokine-alpha is soluble or membrane-bound;   (d) contacting the sample with a second anti-Neutrokine-alpha antibody, which binds to soluble Neutrokine-alpha;   (e) detecting binding of the first antibody and the second antibody to Neutrokine-alpha in the sample; and   (f) comparing the binding of the first antibody and the second antibody in the biological sample to the binding of the first and second antibodies in a control sample,   wherein a difference in binding attributable to the test compound indicates that the compound or composition is effective in modulating the distribution of Neutrokine-alpha in the mammal.   
     
     
         49 . The method of  claim 46 , wherein the biological sample or samples is or are obtained from a mammal that has an autoimmune disease. 
     
     
         50 . The method of  claim 46 , wherein the binding is evaluated qualitatively or quantitatively. 
     
     
         51 . The method of  claim 46 , wherein
 (a) the biological sample or samples comprises or comprise serum and the method comprises ELISA,   (b) the biological sample or samples comprises or comprise blood and the method comprises flow cytometry, or   (c) the biological sample or samples comprises or comprise tissue and the method comprises immunohistochemistry.   
     
     
         52 . The method of  claim 46 , wherein the biological sample or samples comprises or comprise serum, blood, plasma, biopsy, tissue, suspension cell line, body fluid, spinal fluid, cells of B cell lineage, urine, or synovial fluid. 
     
     
         53 . The method of  claim 47 , wherein the binding of the first anti-Neutrokine-alpha antibody indicates total Neutrokine-alpha. 
     
     
         54 . The method of  claim 47 , wherein the binding of the second anti-Neutrokine-alpha antibody indicates free Neutrokine-alpha. 
     
     
         55 . The method of  claim 47 , wherein the difference in binding of the first and second antibodies indicates receptor-bound Neutrokine-alpha in the sample. 
     
     
         56 . An isolated anti-Neutrokine-alpha antibody selected from the group consisting of:
 (a) an antibody that binds Neutrokine-alpha ligand and Neutrokine-alpha ligand-receptor complexes, whether the Neutrokine-alpha is soluble or membrane-bound; and   (b) an antibody that binds Neutrokine-alpha ligand and does not prevent the ligand from binding a Neutrokine-alpha receptor.   
     
     
         57 . The antibody of  claim 56 , wherein the affinity of the antibody for free Neutrokine-alpha and the affinity of the antibody for Neutrokine-alpha bound to Neutrokine-alpha receptor differ. 
     
     
         58 . The antibody of  claim 56 , wherein the antibody binds to Neutrokine-alpha bound to Neutrokine-alpha receptor with an affinity of at least 10 −6  M. 
     
     
         59 . A method of selecting the antibody of  claim 56 , comprising determining whether the antibody binds Neutrokine-alpha independent of whether Neutrokine-alpha is bound to a receptor. 
     
     
         60 . A method of using the antibody of  claim 56 , comprising contacting a biological sample with the antibody, thereby evaluating the biological sample. 
     
     
         61 . A method of identifying a subject who is a candidate for a Neutrokine-alpha antagonist therapy, the method comprising:
 (a) contacting a biological sample from the subject with an anti-Neutrokine-alpha antibody that binds Neutrokine-alpha ligand and Neutrokine-alpha ligand-receptor complexes, whether the Neutrokine-alpha is soluble or membrane-bound;   (b) evaluating the level of total Neutrokine-alpha bound by the antibody; and   (c) selecting a subject who has a greater level of total Neutrokine-alpha bound by the antibody, compared to a control,   wherein a greater level of total Neutrokine-alpha bound by the antibody, compared to a control, indicates that the subject is a candidate for a Neutrokine-alpha antagonist therapy.   
     
     
         62 . The method of  claim 61 , wherein the Neutrokine-alpha antagonist therapy is selected from the group consisting of: an inhibitory anti-Neutrokine-alpha antibody, an antagonistic anti-Neutrokine-alpha antibody, a neutralizing anti-Neutrokine-alpha antibody, and an anti-Neutrokine-alpha receptor antibody. 
     
     
         63 . The method of  claim 61 , wherein the subject has an autoimmune disease selected from the group consisting of: autoimmune hemolytic anemia, autoimmune neonatal thrombocytopenia, idiopathic thrombocytopenia purpura, autoimmunocytopenia, hemolytic anemia, antiphospholipid syndrome, dermatitis, allergic encephalomyelitis, myocarditis, relapsing polychondritis, rheumatic heart disease, glomerulonephritis, IgA nephropathy, Multiple Sclerosis, Neuritis, Uveitis Ophthalmia, Polyendocrinopathies, Purpura, Henloch-Scoenlein purpura, Reiter's Disease, Stiff-Man Syndrome, Autoimmune Pulmonary Inflammation, Guillain-Barre Syndrome, insulin dependent diabetes mellitus, autoimmune inflammatory eye disease, autoimmune thyroiditis, hypothyroidism, Hashimoto's thyroiditis, systemic lupus erythematosus, Goodpasture's syndrome, Pemphigus, Receptor autoimmunities, Graves' Disease, Myasthenia Gravis, insulin resistance, autoimmune thrombocytopenic purpura, rheumatoid arthritis, scleroderma with anti-collagen antibodies, mixed connective tissue disease, polymyositis/dermatomyositis, pernicious anemia, idiopathic Addison's disease, infertility, glomerulonephritis, primary glomerulonephritis, bullous pemphigoid, Sjogren's syndrome, diabetes mellitus, adrenergic drug resistance, adrenergic drug resistance with asthma or cystic fibrosis, chronic active hepatitis, primary biliary cirrhosis, other endocrine gland failure, vitiligo, vasculitis, post-MI, cardiotomy syndrome, urticaria, atopic dermatitis, asthma, and inflammatory myopathies. 
     
     
         64 . The method of  claim 61 , wherein the biological sample is selected from the group consisting of: blood, tissue, plasma, synovial fluid, and spinal fluid. 
     
     
         65 . The method of  claim 61 , wherein the subject has a normal level of free soluble Neutrokine-alpha compared to the control. 
     
     
         66 . A method of identifying a subject who is a candidate for a Neutrokine-alpha antagonist therapy, the method comprising:
 (a) contacting a biological sample from the subject with a first anti-Neutrokine-alpha antibody, which binds Neutrokine-alpha ligand and Neutrokine-alpha ligand-receptor complexes, whether the Neutrokine-alpha is soluble or membrane-bound;   (b) contacting the biological sample with a second anti-Neutrokine-alpha antibody, which binds to soluble Neutrokine-alpha; and   (c) comparing the binding of the first antibody and the second antibody, thereby evaluating the level of receptor-bound Neutrokine-alpha,   wherein a greater level of receptor-bound Neutrokine-alpha, compared to a control, indicates that the subject is a candidate for a Neutrokine-alpha antagonist therapy.   
     
     
         67 . The method of  claim 66 , wherein the Neutrokine-alpha antagonist therapy is selected from the group consisting of: an inhibitory anti-Neutrokine-alpha antibody, an antagonistic anti-Neutrokine-alpha antibody, a neutralizing anti-Neutrokine-alpha antibody, and an anti-Neutrokine-alpha receptor antibody. 
     
     
         68 . The method of  claim 66 , wherein the subject has an autoimmune disease selected from the group consisting of: autoimmune hemolytic anemia, autoimmune neonatal thrombocytopenia, idiopathic thrombocytopenia purpura, autoimmunocytopenia, hemolytic anemia, antiphospholipid syndrome, dermatitis, allergic encephalomyelitis, myocarditis, relapsing polychondritis, rheumatic heart disease, glomerulonephritis, IgA nephropathy, Multiple Sclerosis, Neuritis, Uveitis Ophthalmia, Polyendocrinopathies, Purpura, Henloch-Scoenlein purpura, Reiter's Disease, Stiff-Man Syndrome, Autoimmune Pulmonary Inflammation, Guillain-Barre Syndrome, insulin dependent diabetes mellitus, autoimmune inflammatory eye disease, autoimmune thyroiditis, hypothyroidism, Hashimoto's thyroiditis, systemic lupus erythematosus, Goodpasture's syndrome, Pemphigus, Receptor autoimmunities, Graves' Disease, Myasthenia Gravis, insulin resistance, autoimmune thrombocytopenic purpura, rheumatoid arthritis, scleroderma with anti-collagen antibodies, mixed connective tissue disease, polymyositis/dermatomyositis, pernicious anemia, idiopathic Addison's disease, infertility, glomerulonephritis, primary glomerulonephritis, bullous pemphigoid, Sjogren's syndrome, diabetes mellitus, adrenergic drug resistance, adrenergic drug resistance with asthma or cystic fibrosis, chronic active hepatitis, primary biliary cirrhosis, other endocrine gland failure, vitiligo, vasculitis, post-MI, cardiotomy syndrome, urticaria, atopic dermatitis, asthma, and inflammatory myopathies. 
     
     
         69 . The method of  claim 66 , wherein the biological sample is selected from the group consisting of: blood, tissue, plasma, synovial fluid, and spinal fluid. 
     
     
         70 . The method of  claim 66 , wherein the subject has a normal level of free Neutrokine-alpha compared to the control. 
     
     
         71 . A method of monitoring a patient who has been administered a therapy for a B cell mediated disease, the method comprising performing the method of  claim 46  on a biological sample from the patient, wherein a decrease in total Neutrokine-alpha in the biological sample indicates that the patient is responsive to the therapy. 
     
     
         72 . The method of  claim 71 , wherein the method is performed at least one month after the administration of the therapy. 
     
     
         73 . The method of  claim 71 , wherein the method is repeated after the administration of the therapy. 
     
     
         74 . The method of  claim 71 , wherein the subject has an autoimmune disease selected from the group consisting of: autoimmune hemolytic anemia, autoimmune neonatal thrombocytopenia, idiopathic thrombocytopenia purpura, autoimmunocytopenia, hemolytic anemia, antiphospholipid syndrome, dermatitis, allergic encephalomyelitis, myocarditis, relapsing polychondritis, rheumatic heart disease, glomerulonephritis, IgA nephropathy, Multiple Sclerosis, Neuritis, Uveitis Ophthalmia, Polyendocrinopathies, Purpura, Henloch-Scoenlein purpura, Reiter's Disease, Stiff-Man Syndrome, Autoimmune Pulmonary Inflammation, Guillain-Barre Syndrome, insulin dependent diabetes mellitus, autoimmune inflammatory eye disease, autoimmune thyroiditis, hypothyroidism, Hashimoto's thyroiditis, systemic lupus erythematosus, Goodpasture's syndrome, Pemphigus, Receptor autoimmunities, Graves' Disease, Myasthenia Gravis, insulin resistance, autoimmune thrombocytopenic purpura, rheumatoid arthritis, scleroderma with anti-collagen antibodies, mixed connective tissue disease, polymyositis/dermatomyositis, pernicious anemia, idiopathic Addison's disease, infertility, glomerulonephritis, primary glomerulonephritis, bullous pemphigoid, Sjogren's syndrome, diabetes mellitus, adrenergic drug resistance, adrenergic drug resistance with asthma or cystic fibrosis, chronic active hepatitis, primary biliary cirrhosis, other endocrine gland failure, vitiligo, vasculitis, post-MI, cardiotomy syndrome, urticaria, atopic dermatitis, asthma, and inflammatory myopathies. 
     
     
         75 . The method of  claim 71 , wherein the biological sample is selected from the group consisting of: blood, tissue, plasma, synovial fluid, and spinal fluid. 
     
     
         76 . The method of  claim 72 , wherein the subject has a normal level of free Neutrokine-alpha compared to the control.