Shinobu Matsui

Dilated cardiomyopathy defines serum autoantibodies against G-protein-coupled cardiovascular receptors

In order further to identify the prevalence of anti-receptor autoantibodies in the sera of patients with dilated cardiomyopathy (DCM), we attempted to detect autoantibodies against a series of G-protein-coupled cardiovascular receptors in a well-defined population of DCM patients from Japan. Peptides corresponding to the sequences of the second extracellular loops of the human beta 1 and beta 2 adrenoceptors, alpha 1 adrenoceptors, M2 muscarinic acetylcholine receptors and angiotensin II-1 (AT1) receptors were used as antigens in an enzyme immunoassay to screen the sera from patients with DCM (n = 28). Nine sera from patients with DCM (32%) and 2 sera from healthy subjects (9%) recognized the beta 1 adrenoceptor peptide. Ten sera from patients (36%) and 3 sera from healthy subjects (13%) recognized the M2 receptor peptide. Thirty-six per cent of the patients with autoantibody against the beta 1 adrenoceptor peptide. Ten sera from patients (36%) and 3 sera from healthy subjects (13%) recognized the M2 receptor peptide. Thirty-six per cent of the patients with autoantibody against the beta 1 adrenoceptor had autoantibody against the M2 receptor. However, no significantly high frequencies of autoantibodies against the beta 2 adrenoceptor, alpha 1 adrenoceptor and AT1 receptor were found in DCM patients. Our results demonstrate that a subgroup of patients with DCM have a specific spectrum of autoantibodies which are specifically directed against the second extracellular loops of the beta 1 adrenoceptors and M2 muscarinic receptors rather than other cardiovascular receptors.

Assessment of working skeletal muscle oxygenation in patients with chronic heart failure

Patients with chronic heart failure (CHF) are frequently limited by muscle fatigue resulting from impaired skeletal muscle blood flow. Accordingly, we assessed working skeletal muscle oxygenation in such patients using near-infrared (NIR) spectroscopy. Nine normal subjects (mean age 52 years) and 12 patients with CHF (mean age 60 years) were studied. NIR spectroscopy was used to monitor relative changes in oxygenated hemoglobin (Hb) and myoglobin (Mb) (oxy Hb/Mb), deoxygenated Hb and Mb (deoxy Hb/Mb), and total (oxy + deoxy) Hb and Mb (total Hb/Mb) contents in the vastus lateralis muscle at rest, during warm-up (0 W, 30 cycles/min for 3 min), incremental maximal supine bicycle exercise (ramp protocol, 15 W/min, 50 cycles/min), and recovery. At peak exercise the patients exhibited reduced heart rate, systolic blood pressure, peak exercise oxygen consumption (VO2; 15 +/- 3.0 ml/kg/min vs 32 +/- 8.5 ml/kg/min), and workload (99 +/- 23.4 W vs 183 +/- 68.4 W) as compared with the normal subjects. The respiratory quotient was comparable in both groups. In the normal subjects, oxy Hb/Mb was increased from the warm-up period to the early phase of exercise, followed by a progressive decrease to peak exercise. In the recovery phase, oxy Hb/Mb was increased abruptly. For these patients, change in oxy Hb/Mb followed a pattern similar to that seen in normal subjects, and oxy Hb/Mb was decreased earlier in contrast to that in the normal subjects. There was a significant difference in the change of oxy Hb/Mb during warm-up, early phase exercise, and recovery between the two groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Autoantibodies against M2 muscarinic receptors in patients with cardiomyopathy display non-desensitized agonist-like effects.

Circulating autoantibodies against the human M2 muscarinic receptors have been previously shown in 38% of patients with idiopathic dilated cardiomyopathy. The functional properties of these autoantibodies are reported herein. They were able to decrease the cell beating frequency of myocytes in cultured neonatal rat heart cells in a dose-dependent manner without desensitization over a period of more than 5 hours whereas the non-specific muscarinic receptor agonist carbachol also inhibited the heart cell beating frequency but was desensitized within 1 hour. In the same cell culture, anti-M2 muscarinic receptor autoantibodies were not able to induce internalization of muscarinic receptor whereas carbachol did. These results demonstrate for the first time that anti-M2 muscarinic receptor autoantibodies from patients with idiopathic dilated cardiomyopathy have stimulatory muscarinic activity in vitro, which differ from normal muscarinic agonists by non-desensitization.

Active immunization of combined β1-adrenoceptor and M2-muscarinic receptor peptides induces cardiac hypertrophy in rabbits

BACKGROUND The high prevalence of patients with dilated cardiomyopathy (DCM) with anti-beta1-adrenoceptor and/or anti-M2-muscarinic receptor autoantibodies in their sera has been observed. However, the pathophysiological role of these autoantibodies in the development of cardiomyopathy is unknown. We previously reported an experimental model of early-stage DCM-like cardiomyopathy induced by immunizing rabbits for 1 year with synthetic peptides corresponding to the sequence of the second extracellular loop of either beta1-adrenoceptor or M2-muscarinic receptor. Because approximately half the sera of patients with DCM that recognize one of the two receptor sequences also recognize the second sequence, a model was created in rabbits simultaneously immunized with the synthetic peptides corresponding to the second extracellular loop of the beta1-adrenoceptor and M2-muscarinic receptor. METHODS AND RESULTS All rabbits (n = 8) immunized with both peptides had a high titer of both anti-beta1-adrenoceptor and anti-M2-muscarinic receptor autoantibodies in their sera, whereas none of the sera from control rabbits injected with saline (n = 9) was positive. No significant cross-reaction with peptides other than those used for immunization was found. The weight of the hearts of immunized rabbits increased significantly. The hearts of immunized rabbits showed marked concentric left ventricular hypertrophy with mild inflammatory cell infiltration. In these rabbits, mild or moderate interstitial fibrosis was also observed. In electron micrographs, immunized rabbits showed focal myofibrillar lysis, loss of myofilament, and a marked increase in the number of mitochondria and deposition of dense granules in both sarcoplasm and myofibrils. Conversely, one of the control rabbits showed scant mononuclear cell infiltration. However, in this control rabbit, no significant alteration was found by electron microscopy. CONCLUSION Our results showed the coexistence of both anti-beta1-adrenoceptor and anti-M2-muscarinic receptor autoantibodies in the sera has pathophysiological importance, shown by their ability to induce cardiac hypertrophy in rabbits.

Exercise blood pressure in young adults as a predictor of future blood pressure: a 12-year follow-up of medical school graduates.

It has not been fully clarified whether exercise blood pressure (BP) in young adult men and women is useful to predict future BP, especially in Asian people. A long-term prospective study was conducted in graduates of a medical school in Japan; 138 men and 76 women whose mean age was 19.8 and 19.2, respectively, at baseline. A 5-min exercise tolerance test was performed at baseline, and BP immediately after exercise was measured. BP at 50% intensity exercise was also calculated. Multiple regression analysis was carried out to clarify the relationship of exercise BP at baseline to follow-up BP after an average of 12 years. In multivariate-adjusted models, the relationship of systolic blood pressure (SBP) at follow-up was stronger to SBP immediately after exercise (F=7.7, P=0.006) than to resting SBP (F=3.7, P=0.055) in men. The models in men showed that SBP immediately after exercise was a stronger predictor of follow-up SBP than SBP at 50% intensity exercise, and the results were similar for diastolic blood pressure (DBP) in men. For SBP in women, resting SBP was the strongest predictor of follow-up SBP (F=14.3, P<0.001), and exercise SBP was not significant predictor. For DBP in women, any DBP at rest or after exercise was not significantly related to DBP at follow-up. In young adult men, SBP and DBP immediately after exercise would be a stronger predictor of future SBP and DBP rather than BP at rest. However, in young adult women, resting SBP rather than exercise SBP would be better to predict future SBP.

Transfer of rabbit autoimmune cardiomyopathy into severe combined immunodeficiency mice.

Summary: Growing evidence suggests that the autoimmune mechanism plays an important role in the pathogenesis of dilated cardiomyopathy. The purpose of this study was to evaluate the effect on the cardiac structure and function by the transfer of immunoglobulin G (IgG) and/or lymphocytes from rabbits immunized with a synthetic peptide corresponding to the sequence of the second extracellular loop of &bgr;1‐adrenoceptor (&bgr; peptide) into severe combined immunodeficiency (SCID) mice. CB‐17 SCID mice were injected intraperitoneally with 2 mg of IgG and/or 1 × 107 peripheral blood lymphocytes (PBL) from either rabbits immunized with both &bgr;1 peptide and adjuvant (&bgr; group), and adjuvant or rabbits with adjuvant only (N group). Thirty‐five SCID mice were divided into seven groups: (1) N‐IgG group; (2) N‐PBL group; (3) N‐IgG+PBL group; (4) &bgr;‐IgG group; (5) &bgr;‐PBL group; (6) &bgr;‐IgG+PBL group; and (7) control group. Morphological, serological and endocrinological studies were performed 70 days after the transfer. Results showed that heart weight and heart weight/body weight ratio in the &bgr;‐IgG+PBL group tended to be increased as compared with those in other groups. All mice in the &bgr;‐IgG group, two in the &bgr;‐PBL group and four in the &bgr;‐IgG+PBL group showed high titer of rabbit anti‐ &bgr;1‐adrenoceptor antibodies. Brain natriuretic peptide in the &bgr;‐IgG+PBL group showed a significant increase as compared with those in the control group and N‐IgG+PBL. Pathohistologically, focal infiltration of inflammatory cells in the myocardium was observed in one mouse of the &bgr;‐IgG+PBL group. Rabbit CD3‐positive T‐lymphocytes in the myocardium were observed in two mice of the &bgr; group. In conclusion, transfer of IgG and PBL from rabbits immunized with &bgr;1 peptide was able to induce the early stages of myocardial damage in SCID mice. These data provide direct evidence that the autoimmune mechanism is important in the pathogenesis of dilated cardiomyopathy.

Hemodynamic effects of dibutyryl cyclic AMP in congestive heart failure.

To evaluate the hemodynamic effects of dibutyryl cyclic AMP (DBcAMP) in congestive heart failure (CHF), right-sided cardiac catheterization was performed in 11 patients with CHF, and hemodynamic variables were investigated before and after infusion of various doses of DBcAMP at a rate of 0.025 to 0.2 mg/kg/min (mean 0.14 +/- 0.077 [standard deviation]). DBcAMP reduced total systemic vascular resistance index from 3,171 +/- 1,158 to 1,880 +/- 554 dynes s cm-5 X m2 (mean +/- standard deviation) and pulmonary arterial end-diastolic pressure from 23 +/- 13 to 20 +/- 11 mm Hg, and increased cardiac index from 2.24 +/- 0.60 to 3.41 +/- 1.02 liters/min/m2. Mean arterial blood pressure decreased from 91 +/- 14 to 84 +/- 13 mm Hg, and heart rate increased from 91 +/- 16 to 99 +/- 13 beats/min. The increase in cardiac index was accompanied by a proportional decrease in total systemic vascular resistance index in all patients except 1. In 8 patients the decrease in pulmonary arterial end-diastolic pressure was accompanied by an increase or no change in the left ventricular stroke work index. In 6 patients, DBcAMP was given in incremental doses of 0.05, 0.1, and 0.2 mg/kg/min every 20 minutes, and 5 of 6 patients tolerated the full dose and showed dose-related hemodynamic changes for the incremental doses of DBcAMP. These data suggest that DBcAMP has powerful vasodilating effects on resistance vessels in patients with CHF; hence, it can be a useful vasodilating agent for treatment of CHF.

Characteristic distribution of circulating autoantibodies against G-protein coupled cardiovascular receptors in patients with idiopathic dilated and hypertrophic cardiomyopathy

Circulating autoantibodies against cardiovascular membrane receptors have been repeatedly demonstrated in a subgroup of sera of patients with idiopathic dilated cardiomyopathy. This short review summarizes the clinical findings on anti-receptor autoantibodies in patients with dilated cardiomyopathy.

Transfer of immune components from rabbit autoimmune cardiomyopathy into severe combined immunodeficiency (SCID) mice induces cardiomyopathic changes

Background: Growing evidence suggests that autoimmune mechanism plays an important role in the pathogenesis of cardiomyopathy. The purpose of this study was to investigate whether passive transfer of IgG and/or lymphocytes from rabbits with autoimmune cardiomyopathy is able to reproduce cardiomyopathic changes in severe combined immunodeficiency (SCID) mice. Methods and results: SCID mice were injected intraperitoneally with IgG and/or peripheral blood lymphocytes (PBL) from either rabbits immunized with both β1-adrenoceptor peptide and M2-muscarinic receptor peptide (β1+M2 group) or rabbits with adjuvant (N group). Thirty five SCID mice were divided into seven groups; N-IgG, N-PBL, N-IgG & PBL, (β1+M2)-IgG, (β1+M2)-PBL, (β1+M2)-IgG & PBL and control groups. Heart weight in three (β1+M2) groups were significantly increased. All mice in three (β1+M2) groups showed high titer of rabbit anti-β1 adrenocepter autoantibodies, and 4 mice in the (β1+M2)-PBL group and 3 mice in the (β1+M2)-IgG & PBL group showed a significant increase in titer of rabbit anti-M2-muscarinic receptor autoantibodies. Focal infiltration of inflammatory cells in the myocardium was observed in the (β1+M2)-IgG & PBL group. In the (β1+M2)-PBL group and (β1+M2)-IgG & PBL group, cardiomyocyte diameters were significantly increased. Some myocytes of the (β1+M2)-IgG & PBL group exhibited intracellular edema, clumps of Z-band and increased numbers of mitochondria by using electron microscopy. Conclusion: Transfer of IgG and PBL from rabbits immunized with combined β1 and M2 peptides was able to reproduce the early stage of cardiomyopathic changes in SCID mice

Amiodarone minimizes experimental autoimmune myocarditis in rats

Amiodarone, a promising drug for the treatment of tachyarrythmias, was recently found to have immunomodulatory effects in vitro. We hypothesized that amiodarone would affect the immune system in vivo and examined the effect of amiodarone on myocarditis in rats. We induced experimental autoimmune myocarditis in rats by cardiac myosin immunization and treated the animals with an intraperitoneal injection of amiodarone at 25 mg/kg/every other day, 10 times after the induction of experimental autoimmune myocarditis. In the treated group, both microscopic and macroscopic examinations showed reduced heart weights, a mild and localized infiltration of inflammatory cells and fibrosis in the myocardium, and a mild congestion in the liver and lungs as compared with the control group. The phenotypic distribution of lymphocytes in peripheral blood showed a significant decrease in the CD4/CD8a ratio in the treated group, but not in the control group. The proportion of mast cells involved in inflammatory cell infiltration was lower in the treated group than the control group. In vitro, amiodarone inhibited the proliferation of mast cells by arresting them in the G2 phase of the cell cycle. These results indicated that amiodarone minimized the progression of experimental autoimmune myocarditis, suggesting a potential therapeutic role for amiodarone treatment in patients suffering from myocarditis, especially myocarditis complicated by cardiac arrhythmias. One possible mechanism by which amiodarone minimizes the progression of experimental autoimmune myocarditis may be to affect the immune system via the immunomodulatory effects on T cell and mast cell functions.