METHODS AND COMPOSITIONS RELATED TO ACETYLOCHOLINE RECEPTOR CONJUGATES

• US 20110237688A1
• Filed: 07/29/2009
• Published: 09/29/2011
• Est. Priority Date: 07/29/2008
• Status: Active Grant

• Current Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Original Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Inventors: Allman, Windy, Christadoss, Premkumar
• Granted Patent: US 8,530,245 B2
• Time in Patent Office: Days
• US Class Current: 514/789
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: G01N 2333/70571 for neuromediators, e.g. serotonin receptor, dopamine receptor
• CPC Class Codes: G01N 2800/102 Arthritis; Rheumatoid arthritis, i.e. inflammation of peripheral joints
• CPC Class Codes: G01N 2800/104 Lupus erythematosus [SLE]
• CPC Class Codes: G01N 33/564 for pre-existing immune complex or autoimmune disease , i.e. systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, rheumatoid factors or complement components C1-C9
• CPC Class Codes: G01N 33/56972 White blood cells

Abstract

Disclosed are methods, compositions, and diagnostic kits for detecting acetylcholine receptor (AchR) autoreactive immune cells in a subject. The methods comprise detecting the binding of AChR-conjugate to penpheral blood AChR-specific B cells for diagnosing autoimmune disorders, including Myasthenia gravis (MG), systemic lupus erythematous (SLE), and rheumatoid arthritis (RA). More specifically, the detection is achieved by using flow cytometric assay with Alexa-conjugated AchR.

Claims

1. A method of evaluating a patient for or with Myasthenia Gravis comprising the step of quantitating acetylcholine receptor (AChR) conjugate binding to B cells in a sample from the patient.

2. The method of claim 1, wherein AChR conjugate binding is evaluated with respect to a reference or standard.

3. The method of claim 1, wherein the sample is a blood sample.

4. The method of claim 1, wherein the AChR conjugate comprises a fluorophore.

5. The method of claim 4, wherein the fluorophore is an Alexa fluorophore.

6. The method of claim 5, wherein the Alexa fluorophore is Alexa-488 or Alexa-647.

7. The method of claim 1, wherein determining a level of AChR binding B cells is by flow cytometry.

8. The method of claim 1, further comprising contacting the sample with B cell marker binding agent.

9. The method of claim 8, wherein the B cell marker binding agent is an antibody.

10. The method of claim 9, wherein the marker is a cell surface molecule.

11. The method of claim 9, wherein the antibody binds IgG, CD19, CD21, CD45R, CD20, CD22, CD23, or CD81.

12. The method of claim 1, further comprising administering a treatment for myasthenia gravis.

13. A device for evaluating a patient for myasthenia gravis comprising an AChR conjugate.

14. An acetylcholine receptor conjugate comprising at least 10 consecutive amino acids of an AChR polypeptide coupled to a detectable moiety.

15. A kit for detecting acetylcholine receptor reactive B cells comprising an acetylcholine receptor conjugate.

16. A method of evaluating a patient for or with an autoimmune condition comprising the steps of:

(i) contacting a sample comprising B cells with a protein or peptide conjugate that specifically binds a B cell associated with the autoimmune condition;

(ii) determining a level of conjugate binding B cells in a sample; and

(iii) comparing the level of conjugate reactive B-cells with a reference or standard.

17. The method of claim 16, wherein the autoimmune condition is SLE or rheumatoid arthritis.