Proinflammatory cytokines and their receptors are increased in the peripheral blood of patients with heart failure. We measured cytokines and their receptors in systemic artery (SA), coronary sinus (CS) and infra-renal inferior vena cava (IVC), in order to investigate their origin and influential factors. Thirty patients with idiopathic dilated cardiomyopathy were performed echocardiography at admission, and right heart catheterization after stabilization. Blood was drawn from 3 sites for measurement of tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6) and soluble tumor necrosis factor- receptor (sTNFR) I, II. TNF-alpha at CS (3.25+/-0.34 pg/mL) was higher than those of SA (1.81+/-0.39 pg/mL) and IVC (1.88+/-0.38 pg/mL, p<0.05). IL-6 at CS (18.3+/-3.8 pg/mL) was higher than that of SA (5.8+/-1.2 pg/mL, p<0.01). The levels of sTNFR I, II showed increasing tendency in sequence of SA, IVC and CS. TNF-alpha and sTNFR I, II from all sites were proportional to worsening of functional classes at admission (p<0.05). E/Ea by Doppler study at admission, which reflects left ventricular end-diastolic pressure (LVEDP) was positively correlated with TNF-alpha from SA (R=0.71, p<0.01), CS (R=0.52, p<0.05) and IVC (R=0.46, p<0.05). Thus, elevated LVEDP during decompensation might cause cytokine release from myocardium in patients with idiopathic dilated cardiomyopathy.
Adult ; Aged ; Cardiomyopathy, Congestive/*blood/*immunology ; Female ; Heart/anatomy & histology ; Hemodynamic Processes ; Human ; Interleukin-6/*blood ; Male ; Middle Aged ; Receptors, Tumor Necrosis Factor/*blood ; Statistics ; Tumor Necrosis Factor/*metabolism

Preparation of a pure autoantigen by way of recombinant DNA technology has an important value in an accurate diagnosis or prognosis of an autoimmune disease. BCOADC-E2 subunit, a mitochondrial protein, has been known to be the autoantigen of primary biliary cirrhosis (PBC), a chronic autoimmune liver disease, as well as idiopathic dilated cardiomypathy (IDCM), a chronic autoimmune heart disease. Recombinant form of this molecule had been expressed in E. coli but with low yield and severe degradation. Furthermore, sera from IDCM patients failed to recognized BCOADC-E2 molecule produced in prokaryotic expression system. In this study, a recombinant bovine BCOADC-E2 fusion protein has been expressed in insect cells using baculovirus expression system and analyzed anti-BCOADC-E2 reactivity in sera from patients with PBC or with IDCM. Optimal production of the recombinant fusion protein has been achieved at 20 multiplicity of infection (MOI), and the protein was affinity-purified using metal-binding resins. The affinity-purified BCOADC-E2 protein was successfully recognized by sera from PBC patients, but not by sera from IDCM patients suggesting that the different auto-immune response against BCOADC-E2 is needed to be elucidated in terms of epitope recognition.
Acetyltransferases/metabolism ; Acetyltransferases/immunology ; Acetyltransferases/genetics* ; Animal ; Baculoviridae/genetics ; Cardiomyopathy, Congestive/immunology ; Cattle ; Human ; Immune Sera ; Insects/cytology* ; Ketone Oxidoreductases/metabolism ; Ketone Oxidoreductases/immunology* ; Ketone Oxidoreductases/genetics* ; Liver Cirrhosis, Biliary/immunology ; Multienzyme Complexes/metabolism ; Multienzyme Complexes/immunology* ; Multienzyme Complexes/genetics* ; Protein Engineering/methods ; Recombinant Proteins/isolation & purification ; Recombinant Proteins/immunology ; Recombinant Proteins/genetics*