Autoimmune Conditions and NADPH Oxidase Defects

Abstract

The invention relates to methods and materials involved in diagnosing and treating autoimmune conditions. In particular, the invention relates to methods and materials involved in diagnosing arthritis conditions that are accompanied by an NADPH oxidase deficiency, methods and materials involved in treating, preventing, or delaying the onset of arthritis conditions that are accompanied by an NADPH oxidase deficiency, and methods and materials involved in identifying agonists and antagonists of NADPH oxidase activity.

1 . A method for identifying an agent that enhances NADPH oxidase activity in a cell, wherein said cell is from a non-human animal susceptible to arthritis induction, said method comprising:
 (a) contacting said cell with an NADPH oxidase activator and a test agent to form a test cell,   (b) determining the level of NADPH oxidase activity in said test cell,   (c) determining whether or not said level is greater than a control level of NADPH oxidase activity, wherein said control level is the amount of NADPH oxidase activity in a control cell treated with said activator in the absence of said test agent, and   (d) identifying said agent as enhancing NADPH oxidase activity when said level is greater than said control level.   
     
     
         2 . The method of  claim 1 , wherein said cell is a cell from a DA rat. 
     
     
         3 . The method of  claim 1 , wherein said cell is a lymphocyte. 
     
     
         4 . The method of  claim 1 , wherein said non-human animal is susceptible to pristane induced arthritis, collagen induced arthritis, adjuvant induced arthritis, oil induced arthritis, hexadecane induced arthritis, or avridine induced arthritis. 
     
     
         5 . The method of  claim 1 , wherein said NADPH oxidase activator is norfloxacin, phosphatidic acid, diacylglycerol, arachidonic acid, phorbol myristate acetate, lysophosphatidylcholine, fMLP, pristane, phytol, octane, decane, undecane, dodecane, tridecane, tetradecane, hexadecane, heptadecane, octadecane, galectin 1, or galectin 3. 
     
     
         6 . The method of  claim 1 , wherein said level of NADPH oxidase activity is determined by measuring superoxide. 
     
     
         7 . The method of  claim 1 , wherein said level of NADPH oxidase activity is determined by measuring reactive oxygen species. 
     
     
         8 . The method of  claim 1 , wherein said level of NADPH oxidase activity is determined with a cytochrome C assay or WST-1 assay. 
     
     
         9 . A method for diagnosing an autoimmune condition accompanied by NADPH oxidase deficiency in a mammal having an autoimmune condition, said method comprising:
 (a) providing a sample comprising a cell from said mammal;   (b) determining the level of NADPH oxidase activity of said cell after contacting said cell with an NADPH oxidase activator;   (c) determining whether or not said level is less than a control level of NADPH oxidase activity, wherein said control level is the average amount of NADPH oxidase activity of control cells from a population of mammals without said autoimmune condition, and wherein said mammals without said autoimmune condition are from the same species as said mammal; and   (d) identifying said mammal as having said autoimmune condition accompanied by NADPH oxidase deficiency when said level is less than said control level.   
     
     
         10 . The method of  claim 9 , wherein said autoimmune condition is arthritis or multiple sclerosis. 
     
     
         11 . The method of  claim 9 , wherein said autoimmune condition accompanied by NADPH oxidase deficiency is AANOD. 
     
     
         12 . The method of  claim 9 , wherein said NADPH oxidase activator is norfloxacin, phosphatidic acid, diacylglycerol, arachidonic acid, phorbol myristate acetate, lysophosphatidylcholine, fMLP, pristane, phytol, octane, decane, undecane, dodecane, tridecane, tetradecane, hexadecane, hexadecene, heptadecane, octadecane, galectin 1, or galectin 3. 
     
     
         13 . The method of  claim 9 , wherein step (c) comprises determining whether or not said level is between 5 and 75 percent less than said control level, and wherein step (d) comprises identifying said mammal as having said autoimmune condition accompanied by NADPH oxidase deficiency when said level is between 5 and 75 percent less than said control level.