Macrophage migration inhibitory factor (mif) promoter polymorphism in inflammatory disease

Abstract

Describe herein is a novel CATT-tetranucleotide repeat polymorphism at position −817 of the human Mif gene that functionally affects the activity of the Macrophage Inhibitory Factor (MIF) promoter in gene reporter assays. Four genotypes are described which comprise 5, 6, 7, or 8-CATT repeat units. Of these, the 5-CATT allele has the lowest level of basal and stimulated MIF promoter activity in vitro. The presence of the low expressing, 5-CATT repeat allele correlated with low disease severity in a cohort of rheumatoid arthritis patients. Methods, compositions and apparatus for detecting this CATT-tetranucleotide repeat polymorphism at position −817 of the human Mif gene, and for using same for assessing predisposition to severe inflammatory disease, are also disclosed.

1 . A method of diagnosis of severity of a non-infectious inflammatory disease or of a predisposition to severity of a non-infectious inflammatory disease, said method comprising detecting a polymorphism in a human Mif promoter that correlates with an increase or decrease in MIF polypeptide expression, wherein detection of said polymorphism is indicative of the severity of said disease or predisposition to severity of said disease. 
     
     
         2 . The method of  claim 1 , wherein said non-infectious inflammatory disease is selected from the group consisting of autoimmunity, rheumatoid arthritis and graft versus host disease. 
     
     
         3 . The method of  claim 2 , wherein said polymorphism in a human Mif promoter is a CATT-tretranucleotide repeat polymorphism at position −817 of the human Mif gene. 
     
     
         4 . The method of  claim 3 , wherein said CATT-tretranucleotide repeat polymorphism at position −817 of the human Mif gene is selected from the group consisting of 5, 6, 7 and 8 repeat units, and presence of a 5 repeat unit in at least one allele indicates occurrence of or predisposition to low disease severity. 
     
     
         5 . The method of  claim 1 , comprising amplifying said Mif promoter using a PCR technique. 
     
     
         6 . A PCR primer set selected to amplify a region of a human Mif promoter, wherein said PCR primer set is selected from the group consisting of: (i) MIF-F (−1024) and MIF-R (−421); (ii) MIF-F (−441) and MIF-R (+4); (iii) MIFF (−13) and MIF-R (+395); and (iv) MIF-F (+379) and MIF-R (+1043). 
     
     
         7 . A method of detecting a polymorphism in a human Mif promoter region comprising using a primer set of  claim 6 . 
     
     
         8 . An article of manufacture comprising a PCR primer set of  claim 6 . 
     
     
         9 . An isolated nucleic acid molecule comprising a human Mif promoter. 
     
     
         10 . An isolated nucleic acid molecule of  claim 9 , that is a genomic DNA fragment. 
     
     
         11 . An isolated nucleic acid molecule of  claim 10 , wherein said genomic DNA fragment has been amplified from a DNA sample of a human subject. 
     
     
         12 . An isolated nucleic acid molecule of  claim 9 , comprising a portion of a human Mif promoter that comprises a CATT-tretranucleotide repeat polymorphism at position −817 of the human Mif gene. 
     
     
         13 . A method of inflammatory disease therapy comprising screening an individual for severity of a non-infectious inflammatory disease or of a predisposition to severity of a non-infectious inflammatory disease comprising:
 detecting in a human subject a polymorphism in a human Mif promoter that correlates with an increase or decrease in MIF polypeptide expression, wherein detection of said polymorphism is indicative of the severity of said disease or predisposition to severity of said disease; and   treating said human subject to prevent or reduce the severity of said inflammatory disease or to delay the onset of said inflammatory disease.   
     
     
         14 . The method of  claim 13 , comprising treating said human subject by administering an effective amount of at least one agent selected from the group consisting of an MIF inhibitor, an anti-TNFα antibody, an anti-IL1 antibody, and anti-IFN-γ antibody, IL-1RA, a steroid, a glucocorticoid, and IL-10. 
     
     
         15 . The method of  claim 14 , wherein said inflammatory disease is rheumatoid arthritis and said polymorphism in a human Mif promoter is a CATT-tretranucleotide repeat polymorphism at position −817 of the human Mif gene.