Methods and kits for diagnosing or monitoring autoimmune and chronic inflammatory diseases
The present invention relates to compositions and methods for diagnosing, monitoring and/or treating an autoimmune or chronic inflammatory disease. In particular, the present invention provides methods for diagnosing, monitoring and treating an autoimmune disease (e.g., rheumatoid arthritis) or chronic inflammatory disease (e.g., systemic lupus erythematosus) based on detecting or altering (e.g., altering expression or methylation status of) autoimmune or chronic inflammatory disease proteins (e.g., CD70 and CD40L). The present invention also provides kits for detecting methylation status of autoimmune or chronic inflammatory disease proteins (e.g., CD70 and CD40L) and for diagnosing, monitoring and/or treating autoimmune or chronic inflammatory diseases.
1. A method for detecting the methylation status of CD40L in a subject comprising: a) providing a biological sample from a subject, wherein the biological sample comprises CD40L nucleic acid sequence; b) exposing said sample to reagents for detecting the methylation status of CD40L nucleic acid sequence; c) detecting the methylation status of CD40L nucleic acid sequence in said sample; d) comparing the methylation status of CD40L nucleic acid sequence in said sample to the methylation status of CD40L nucleic acid sequence in a control sample; and e) correlating the methylation status of CD40L nucleic acid sequence in said sample with the presence or absence of systemic lupus erythematosus or rheumatoid arthritis.
2. The method of claim 1 , wherein said reagents detect methylation status of the 5′ untranslated region of CD40L.
3. The method of claim 2 , wherein said 5′ untranslated region comprises the 125 to −110 and/or the −350 to −400 region of CD40L.
4. The method of claim 1 , wherein said detecting comprises use of a polymerase chain reaction.
5. The method of claim 4 , wherein said polymerase chain reaction is methylation sensitive.
6. The method of claim 1 , wherein said detecting comprises restriction enzyme digestion.
7. The method of claim 1 , wherein said detecting comprises using a kit comprising reagents sufficient for detecting methylation status of CD40L in a subject.
8. The method of claim 1 , further comprising detecting methylation status of one or more of CD70, perforin, CD11a, CD11c, IgE FCRγ1, and CD30.
9. The method of claim 1 , wherein said detecting comprises an oligonucleotide binding assay.
10. The method of claim 1 , wherein said detecting comprises use of a microarray.
11. The method of claim 1 , wherein said subject is a healthy subject that does not display symptoms of autoimmune or chronic inflammatory disease.
12. The method of claim 1 , wherein said CD40L nucleic acid sequence is DNA.
13. The method of claim 1 , wherein said biological sample is selected from the group consisting of bone marrow, whole blood, serum, plasma, interstitial fluid, urine, cerebrospinal fluid, and tissue.