Cardiovascular autoimmune disease panel and methods of using same

Abstract

Provided herein are diagnostic tests, methods of use, and kits for the assessment and management of cardiovascular autoimmune disease and risk of cardiovascular autoimmune disease. Assay methods of the invention can be employed to identify cardiovascular autoimmune disease, or risk thereof, in subjects who have cardiovascular disease, an autoimmune disease, or who are related to an individual with an autoimmune disease; for testing of a subject that exhibits symptoms of cardiovascular disease, as well as of a subject that is apparently healthy and does not yet exhibit symptoms of cardiovascular disease; and for determining whether a subject having, or at risk for, a cardiovascular disease is a candidate for immunosuppressive therapy or immunoabsorption therapy. The invention also provides kits and kit components useful for performing the methods.

1 . A method for assessing the risk of whether a subject has or might develop a cardiovascular autoimmune disease, wherein said method comprises:
 (a) obtaining a sample from said subject;   (b) measuring the levels of autoantibodies to each of a plurality of different endogenous cardiovascular antigens in said subject's sample;   (c) comparing said levels with the level of the same autoantibodies measured in samples from reference subjects with clinically normal cardiovascular function, or from reference subjects having cardiovascular disease, and   (d) identifying said risk that said subject has or might develop a cardiovascular autoimmune disease based on the comparison in step (c).   
     
     
         2 . The method of  claim 1 , wherein said risk is present when levels of autoantibodies to at least two of said different endogenous cardiovascular antigens:
 (i) are elevated in said subject's sample as compared to in samples from reference subjects with clinically normal cardiovascular function, and/or   (ii) are elevated in or at about the same level in said subject's sample as compared to in samples from reference subjects having cardiovascular disease.   
     
     
         3 . The method of  claim 1 , wherein said risk is further increased when said subject either has an autoimmune disease, or is a first-degree relative of an individual having an autoimmune disease 
     
     
         4 . The method of  claim 1 , wherein said levels of autoantibodies to at least two of said different endogenous cardiovascular antigens are elevated due to the presence of cardiovascular autoimmune disease. 
     
     
         5 . The method of  claim 1 , wherein said levels of autoantibodies to at least two of said different endogenous cardiovascular antigens are elevated due to an increased risk of developing cardiovascular autoimmune disease. 
     
     
         6 . The method of  claim 1 , wherein said subject's sample comprises peripheral blood, serum, plasma, cerebrospinal fluid, urine, or other body fluid sample. 
     
     
         7 . The method of  claim 1 , wherein said cardiovascular autoimmune disease is a disease, disorder or condition selected from the group consisting of myocarditis, cardiomyopathy, and ischemic heart disease. 
     
     
         8 . The method of  claim 1 , wherein said plurality comprises two or more. 
     
     
         9 . The method of  claim 1 , wherein said plurality comprises between two and ten. 
     
     
         10 . The method of  claim 1 , wherein said plurality comprises between five and ten. 
     
     
         11 . The method of  claim 1 , wherein said plurality comprises a number selected from the group consisting of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 15, 17, 18, 19, and 20. 
     
     
         12 . The method of  claim 1 , wherein said different endogenous cardiovascular antigens are selected from the group consisting of α1-adrenoceptor, angiotensin-1 receptor, annexin V, brain natriuretic peptide, cardiac troponins, myosin, tropomyosin, cytoplasmic neutrophils, endothelial receptor of protein C, Factor VIII, grehlin, halogenated protein, nitrated protein, heat shock proteins, myeloperoxidase, placental growth factor, phospholipids, proteinase- 3 , prothrombin, Purkinje fibers, sarcolemmal Na-K-ATPase, β1-adrenoceptor, β2-adrenoceptor, and tissue-type plasminogen activator. 
     
     
         13 . The method of  claim 12 , wherein said different endogenous cardiovascular antigens further comprise an antigen selected from the group consisting of cardiolipin, oxidized LDL, and β-2-glycoprotein-1. 
     
     
         14 . A method for identifying a patient with cardiovascular disease as eligible to receive autoimmune therapy, wherein said method comprises:
 (a) obtaining a sample from said cardiovascular disease patient;   (b) measuring the levels of autoantibodies to each of a plurality of different endogenous cardiovascular antigens in said subject's sample;   (c) comparing said levels with the level of the same autoantibodies measured in samples from reference subjects with clinically normal cardiovascular function, or from reference subjects having autoimmune disease; and   (d) identifying said cardiovascular disease patient as eligible to receive autoimmune therapy based on the comparison in step (c).   
     
     
         15 . The method of  claim 14 , wherein said patient is eligible to receive autoimmune therapy when levels of autoantibodies to at least two of said different endogenous cardiovascular antigens:
 (i) are elevated in said cardiovascular disease patient's sample as compared to in samples from reference subjects with clinically normal cardiovascular function, and/or   (ii) are elevated in or at about the same in said cardiovascular disease patient's sample as compared to in samples from reference subjects having autoimmune disease.   
     
     
         16 . The method of  claim 14 , wherein said cardiovascular disease patient further is eligible to receive autoimmune therapy when said subject either has an autoimmune disease, or is a first-degree relative of an individual having an autoimmune disease 
     
     
         17 . The method of  claim 14 , wherein said levels of autoantibodies to at least two of said different endogenous cardiovascular antigens are elevated due to the presence of cardiovascular autoimmune disease. 
     
     
         18 . The method of  claim 14 , wherein said levels of autoantibodies to at least two of said different endogenous cardiovascular antigens are elevated due to an increased risk of developing cardiovascular autoimmune disease. 
     
     
         19 . The method of  claim 14 , wherein said cardiovascular disease patient's sample comprises peripheral blood, serum, plasma, cerebrospinal fluid, urine, or other body fluid sample. 
     
     
         20 . The method of  claim 14 , wherein said cardiovascular autoimmune disease is a disease, disorder or condition selected from the group consisting of myocarditis, cardiomyopathy, and ischemic heart disease. 
     
     
         21 . The method of  claim 14 , wherein said plurality comprises two or more. 
     
     
         22 . The method of  claim 14 , wherein said plurality comprises between two and ten. 
     
     
         23 . The method of  claim 14 , wherein said plurality comprises between five and ten. 
     
     
         24 . The method of  claim 14 , wherein said plurality comprises a number selected from the group consisting of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 15, 17, 18, 19, and 20. 
     
     
         25 . The method of  claim 14 , wherein said different endogenous cardiovascular antigens are selected from the group consisting of α1-adrenoceptor, angiotensin-1 receptor, annexin V, brain natriuretic peptide, cardiac troponins, myosin, tropomyosin, cytoplasmic neutrophils, endothelial receptor of protein C, Factor VIII, grehlin, halogenated protein, nitrated protein, heat shock proteins, myeloperoxidase, placental growth factor, phospholipids, proteinase-3, prothrombin, Purkinje fibers, sarcolemmal Na-K-ATPase, (β1-adrenoceptor, β2-adrenoceptor, and tissue-type plasminogen activator. 
     
     
         26 . The method of  claim 25 , wherein said different endogenous cardiovascular antigens further comprise an antigen selected from the group consisting of cardiolipin, oxidized LDL, and (β-2-glycoprotein-1. 
     
     
         27 . A method of identifying a subject having, or at risk for developing, a cardiovascular autoimmune disease, the method comprising assaying a sample from the subject for the levels of autoantibodies to each of a plurality of different endogenous cardiovascular antigens, wherein the subject has an autoimmune disease, or the subject is a first-degree relative of an individual having an autoimmune disease, and wherein the presence of elevated levels of autoantibodies to at least two of said different endogenous cardiovascular antigens indicates the presence of, or risk of, said cardiovascular autoimmune disease. 
     
     
         28 . A test kit for assaying a sample for autoantibodies to at least two different endogenous cardiovascular antigens, the test kit comprising means for the detection of levels of autoantibodies to each of a plurality of different endogenous cardiovascular antigens. 
     
     
         29 . The test kit of  claim 28 , wherein said test kit additionally comprises a solid phase and a capture agent affixed to said solid phase, wherein the capture agent is a peptide having a sequence that corresponds to a portion or the entirety of the amino acid sequence of the endogenous cardiovascular antigens selected from the group consisting of α1-adrenoceptor, angiotensin-1 receptor, annexin V, brain natriuretic peptide, cardiac troponins, myosin, tropomyosin, cytoplasmic neutrophils, endothelial receptor of protein C, Factor VIII, grehlin, halogenated protein, nitrated protein, heat shock proteins, myeloperoxidase, placental growth factor, phospholipids, proteinase-3, prothrombin, Purkinje fibers, sarcolemmal Na-K-ATPase, β1-adrenoceptor, β2-adrenoceptor, and tissue-type plasminogen activator. 
     
     
         30 . The test kit of  claim 29 , wherein said different endogenous cardiovascular antigens further comprise an antigen selected from the group consisting of cardiolipin, oxidized LDL, and β-2-glycoprotein-1. 
     
     
         31 . The test kit of  claim 28 , wherein the test kit additionally comprises a labeled detection agent, comprising a species-specific antibody. 
     
     
         32 . The test kit of  claim 31 , additionally comprising an indicator reagent that interacts with said label to produce a detectable signal. 
     
     
         33 . The test kit of  claim 29 , wherein the solid phase comprises a microplate. 
     
     
         34 . The test kit of  claim 29 , wherein the solid phase comprises a microparticle. 
     
     
         35 . The test kit of  claim 29 , wherein the solid phase comprises an electrode. 
     
     
         36 . A method for assessing the risk of whether a subject has or might develop a cardiovascular autoimmune disease, wherein said method comprises:
 (a) obtaining a sample from said subject;   (b) measuring the levels of one or more endogenous cardiovascular antigens in said subject's sample;   (c) measuring the levels of autoantibodies to each of a plurality of different endogenous cardiovascular antigens in said subject's sample;   (d) comparing said levels of said one or more endogenous cardiovascular antigens with the level of the same endogenous cardiovascular antigen measured in samples from reference subjects with clinically normal cardiovascular function, in samples from reference subjects having cardiovascular disease, or in said subject's sample obtained at an earlier time;   (e) comparing said levels of autoantibodies with the level of the same autoantibody measured in samples from reference subjects with clinically normal cardiovascular function, in samples from reference subjects having cardiovascular disease, or in said subject's sample obtained at an earlier time; and   (f) identifying said risk that said subject has or might develop a cardiovascular autoimmune disease based on the comparison in steps (d) and (e).   
     
     
         37 . The method of  claim 36 , wherein said risk is present when:
 (i) in step (d) the levels of said one or more endogenous cardiovascular antigens are altered in said subject's sample as compared to in samples from reference subjects with clinically normal cardiovascular function, and/or are altered in or at about the same level in said subject's sample as compared to in said subject's sample obtained at an earlier time; and/or   (ii) in step (e) the levels of said autoantibodies to at least two of said different endogenous cardiovascular antigens are elevated in said subject's sample as compared to in samples from reference subjects with clinically normal cardiovascular function, and/or are elevated in or at about the same level in said subject's sample as compared to in samples from reference subjects having cardiovascular disease.   
     
     
         38 . The method of  claim 36 , wherein the measuring of step (b) and the measuring of step (c) are done simultaneously. 
     
     
         39 . The method of  claim 36 , wherein the measuring of step (b) and the measuring of step (c) are done sequentially, in any order.