DETECTION AND DIAGNOSIS OF INFLAMMATORY DISORDERS

• US 20090011444A1
• Filed: 08/25/2008
• Published: 01/08/2009
• Est. Priority Date: 12/27/2006
• Status: Active Grant

• Current Assignee: Johns Hopkins University
• Original Assignee: Johns Hopkins University
• Inventors: Chen, Lieping
• Granted Patent: US 7,989,173 B2
• Time in Patent Office: Days
• US Class Current: 435/7.920
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 27/16 Otologicals
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 7/06 Antianaemics
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: G01N 2800/10 Musculoskeletal or connective tissue disorders
• CPC Class Codes: G01N 33/6875 Nucleoproteins

Abstract

Soluble H4 (sH4) levels have been discovered to correlate with the stage or severity of inflammatory disorders including autoimmune disorders. In particular, circulating levels of sH4 can be used as a diagnostic for determining the severity of an inflammatory disorder or the propensity for developing an inflammatory disorder. The severity of an inflammatory disorder can be determined by assaying the levels of sH4 in a subject and comparing the levels of sH4 to reference sH4 concentrations that correlate to specific stages of an inflammatory disorder. The therapeutic efficacy of treatments for inflammatory disorders can also be determined by comparing levels of sH4 before and during treatment. Methods and devices for measuring sH4 are also provided.

Claims

1. A method for determining the severity of an inflammatory disorder comprising determining levels of sH4 in a subject and comparing the levels of sH4 to reference sH4 concentrations that correlate to specific stages of an inflammatory disorder.

2. The method of claim 1 wherein the inflammatory disorder is selected from the group consisting of rheumatoid arthritis, systemic lupus erythematosus, alopecia areata, anklosing spondylitis, antiphospholipid syndrome, autoimmune Addison'"'"'s disease, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune inner ear disease, autoimmune lymphoproliferative syndrome (alps), autoimmune thrombocytopenic purpura (ATP), Behcet'"'"'s disease, bullous pemphigoid, cardiomyopathy, celiac sprue-dermatitis, chronic fatigue syndrome immune deficiency, syndrome (CFIDS), chronic inflammatory demyelinating polyneuropathy, cicatricial pemphigoid, cold agglutinin disease, Crest syndrome, Crohn'"'"'s disease, Dego'"'"'s disease, dermatomyositis, dermatomyositis—

juvenile, discoid lupus, essential mixed cryoglobulinemia, fibromyalgia—

fibromyositis, grave'"'"'s disease, guillain-barre, hashimoto'"'"'s thyroiditis, idiopathic pulmonary fibrosis, idiopathic thrombocytopenia purpura (ITP), Iga nephropathy, insulin dependent diabetes (Type I), juvenile arthritis, Meniere'"'"'s disease, mixed connective tissue disease, multiple sclerosis, myasthenia gravis, pemphigus vulgaris, pernicious anemia, polyarteritis nodosa, polychondritis, polyglancular syndromes, polymyalgia rheumatica, polymyositis and dermatomyositis, primary agammaglobulinemia, primary biliary cirrhosis, psoriasis, Raynaud'"'"'s phenomenon, Reiter'"'"'s syndrome, rheumatic fever, sarcoidosis, scleroderma, Sjogren'"'"'s syndrome, stiff-man syndrome, Takayasu arteritis, temporal arteritis/giant cell arteritis, ulcerative colitis, uveitis, vasculitis, vitiligo, and Wegener'"'"'s granulomatosis.

3. The method of claim 1 wherein the sH4 levels are determined from a fluid sample obtained from the subject.

4. The method of claim 3 wherein the fluid sample is selected from the group consisting of blood, plasma, saliva, lymph, cerebrospinal fluid, synovial fluid, urine, and sputum.

5. The method of claim 1 wherein levels of sH4 are determined using mass spectroscopy, immunohistological methods, immunoprecipitation, an enzyme immunoassay, and a radioimmunoassay.

6. A method for assisting in the diagnosis of an inflammatory disorder or propensity of developing an inflammatory disorder in a subject comprising

obtaining a biological sample from the subject;

determining levels of sH4 in the biological sample, wherein elevated levels of sH4 in the biological sample relative to a control is indicative of an inflammatory disorder or an increased propensity for developing an inflammatory disorder.

7. The method of claim 6 wherein the biological sample is selected from the group consisting of blood, plasma, saliva, lymph, cerebrospinal fluid, synovial fluid, urine, and sputum.

8. The method of claim 6 wherein levels of sH4 are determined using mass spectroscopy, immunohistological methods, immunoprecipitation, an enzyme immunoassay, and a radioimmunoassay.

10. The method of claim 6 wherein the biological sample is selected from the group consisting of blood, plasma, saliva, lymph, cerebrospinal fluid, synovial fluid, urine, and sputum.

11. The method of claim 6 wherein levels of sH4 are determined using mass spectroscopy, immunohistological methods, immunoprecipitation, an enzyme immunoassay, and a radioimmunoassay.

9. A method for determining the efficacy of a treatment for an inflammatory disorder in a subject comprising

obtaining one or more biological samples from the subject during the course of the treatment, and

comparing the levels of sH4 in the samples to the levels of sH4 in a biological sample obtained from the subject prior to treatment, or

comparing the levels of sH4 in the samples to levels of sH4 indicative of different stages of an inflammatory disease or disorder or autoimmune disease.

12. A lateral flow device for determining sH4 concentration in a subject comprising:

(a) an application zone for receiving a fluid sample;

(b) a labeling zone containing labeled binding partner for the sH4;

(c) a detection zone having an immobilized capture reagent for the sH4; and

(d) a reference zone having a signal of fixed intensity indicative of the reference concentration of sH4, wherein when the signal in the detection zone is less intense than the signal in the reference zone, the subject has a sH4 level less than the reference concentration; and

when the signal in the detection zone is more intense than the signal in the reference zone, the subject has a sH4 level more than the reference concentration.

13. The lateral flow device of claim 12 wherein the labeled binding partner of the sH4 is an antibody or antigen binding fragment thereof.

14. The lateral flow device of claim 12 wherein the fluid sample is selected from the group consisting of blood, plasma, saliva, lymph, cerebrospinal fluid, synovial fluid, urine, and sputum.

15. A method for measuring sH4, comprising

(a) obtaining a blood sample from a subject;

(b) applying the blood sample to the application zone of the lateral flow device according to claim 12;

(c) detecting a signal in the detection zone of the lateral flow device; and

(d) comparing the signal detected in step (c) with the signal in the reference zone, wherein when the signal in the detection zone is less intense than the signal in the reference zone, the subject has normal sH4 levels; and

when the signal in the detection zone is equal to or more intense than the signal in the reference zone, the subject has elevated sH4 levels.

18. A kit comprising the lateral flow device of claim 12, reagents for using the kit, and instructions for using the kit.

16. A method for determining neutrophils levels of a subject comprising

obtaining a biological sample from the subject;

determining levels of sH4 in the biological sample;

comparing the levels of sH4 in the biological sample to predetermined levels of sH4 correlating to levels of neutrophils; and

selecting the level of neutrophils that matches the levels of sH4 in the biological sample.

17. The method of claim 16 wherein elevated levels of sH4 relative to a control are indicative of elevated levels of neutrophils relative to a control.