[Show abstract][Hide abstract] ABSTRACT: Recombinant adeno-associated virus (rAAV)-mediated gene transfer is an attractive approach to the treatment of Duchenne muscular dystrophy (DMD). We investigated the muscle transduction profiles and immune responses associated with the administration of rAAV2 and rAAV8 in normal and canine X-linked muscular dystrophy in Japan (CXMD(J)) dogs. rAAV2 or rAAV8 encoding the lacZ gene was injected into the skeletal muscles of normal dogs. Two weeks after the injection, we detected a larger number of beta-galactosidase-positive fibers in rAAV8-transduced canine skeletal muscle than in rAAV2-transduced muscle. Although immunohistochemical analysis using anti-CD4 and anti-CD8 antibodies revealed less T-cell response to rAAV8 than to rAAV2, beta-galactosidase expression in rAAV8-injected muscle lasted for <4 weeks with intramuscular transduction. Canine bone marrow-derived dendritic cells (DCs) were activated by both rAAV2 and rAAV8, implying that innate immunity might be involved in both cases. Intravenous administration of rAAV8-lacZ into the hind limb in normal dogs and rAAV8-microdystrophin into the hind limb in CXMD(J) dogs resulted in improved transgene expression in the skeletal muscles lasting over a period of 8 weeks, but with a declining trend. The limb perfusion transduction protocol with adequate immune modulation would further enhance the rAAV8-mediated transduction strategy and lead to therapeutic benefits in DMD gene therapy.
[Show abstract][Hide abstract] ABSTRACT: Autosomal recessive limb-girdle muscular dystrophy type 2D (LGMD 2D) is caused by mutations in the alpha-sarcoglycan gene (alpha-SG). The absence of alpha-SG results in the loss of the SG complex at the sarcolemma and compromises the integrity of the sarcolemma. To establish a method for recombinant adeno-associated virus (rAAV)-mediated alpha-SG gene therapy into alpha-SG-deficient muscle, we constructed rAAV serotypes 2 and 8 expressing the human alpha-SG gene under the control of the ubiquitous cytomegalovirus promoter (rAAV2-alpha-SG and rAAV8-alpha-SG). We compared the transduction profiles and evaluated the therapeutic effects of a single intramuscular injection of rAAVs into alpha-SG-deficient (Sgca(-/-)) mice. Four weeks after rAAV2 injection into the tibialis anterior (TA) muscle of 10-day-old Sgca(-/-) mice, transduction of the alpha-SG gene was localized to a limited area of the TA muscle. On the other hand, rAAV8-mediated alpha-SG expression was widely distributed in the hind limb muscle, and persisted for 7 months without inducing cytotoxic and immunological reactions, with a reversal of the muscle pathology and improvement in the contractile force of the Sgca(-/-) muscle. This extensive rAAV8-mediated alpha-SG transduction in LGMD 2D model animals paves the way for future clinical application.
Human gene therapy 08/2008; 19(7):719-30. · 4.20 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Using murine models, we have previously demonstrated that recombinant adeno-associated virus (rAAV)-mediated microdystrophin gene transfer is a promising approach to treatment of Duchenne muscular dystrophy (DMD). To examine further therapeutic effects and the safety issue of rAAV-mediated microdystrophin gene transfer using larger animal models, such as dystrophic dog models, we first investigated transduction efficiency of rAAV in wild-type canine muscle cells, and found that rAAV2 encoding beta-galactosidase effectively transduces canine primary myotubes in vitro. Subsequent rAAV2 transfer into skeletal muscles of normal dogs, however, resulted in low and transient expression of beta-galactosidase together with intense cellular infiltrations in vivo, where cellular and humoral immune responses were remarkably activated. In contrast, rAAV2 expressing no transgene elicited no cellular infiltrations. Co-administration of immunosuppressants, cyclosporine and mycophenolate mofetil could partially improve rAAV2 transduction. Collectively, these results suggest that immune responses against the transgene product caused cellular infiltration and eliminated transduced myofibers in dogs. Furthermore, in vitro interferon-gamma release assay showed that canine splenocytes respond to immunogens or mitogens more susceptibly than murine ones. Our results emphasize the importance to scrutinize the immune responses to AAV vectors in larger animal models before applying rAAV-mediated gene therapy to DMD patients.
[Show abstract][Hide abstract] ABSTRACT: The significance of exercise-induced ST segment depression in patients with left circumflex artery involvement was investigated by comparing exercise electrocardiography with exercise thallium-201 single photon emission computed tomography(Tl-SPECT) and the wall motion estimated by left ventriculography. Tl-SPECT and exercise electrocardiography were simultaneously performed in 51 patients with left circumflex artery involvement(angina pectoris 30, myocardial infarction 21). In patients with myocardial infarction, exercise-induced ST depression was frequently found in the V2, V3 and V4 leads. In patients with angina pectoris, ST depression was frequently found in the II, III, aVF, V5 and V6 leads. There was no obvious difference in the leads of ST depression in patients with myocardial infarction with ischemia and without ischemia on Tl-SPECT images. In patients with myocardial infarction, the lateral wall motion of the infarcted area evaluated by left ventriculography was more significantly impaired in the patients with ST depression than without ST depression(p < 0.01). Exercise-induced ST depression in the precordial leads possibly reflects wall motion abnormality rather than ischemia in the lateral infarcted myocardium.
Journal of Cardiology 04/2000; 35(4):239-45. · 2.30 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: To evaluate the regional wall motion and the myocardial fatty acid metabolism at hibernating myocardium after revascularization (PTCA or CABG), we performed dual SPECT with 201Tl and 123I-beta-methyliodophenyl-pentadecanoic acid (BMIPP), and left ventriculography (LVG) in 34 patients with coronary artery disease before and 3 to 4 months after revascularization. In the SPECT, regional tracer uptake was estimated qualitatively (visual) and quantitatively (% uptake). Regional wall motion was estimated qualitatively (visual) and quantitatively (shortening fraction). At the 78 hibernating areas, the improvement of regional wall motion was more significantly (p < 0.05) correlated with that of regional tracer uptake of 123I-BMIPP (r = 0.63) than 201Tl (r = 0.39), and also correlated with the improvement of the difference between 201Tl and 123I-BMIPP regional uptake (r = 0.36). These results suggest that the improvement of wall motion at hibernating myocardium is more significantly correlated with the improvement of 123I-BMIPP than 201Tl uptake after revascularization.
Kaku igaku. The Japanese journal of nuclear medicine 06/1996; 33(6):607-15.
[Show abstract][Hide abstract] ABSTRACT: The association between the lack of adrenergic symptoms during hypoglycemia and myocardial 123I-metaiodobenzylguanidine (MIBG) accumulation was investigated in 12 insulin-treated non-insulin-dependent diabetes mellitus (NIDDM) patients who had no evidence of heart disease. These patients were divided into 2 groups according to the presence (group A) or absence (group B) of adrenergic symptoms during hypoglycemia. Autonomic function tests revealed significantly severe autonomic dysfunction in group B compared to that in group A. Insulin infusion test indicated no significant difference in the catecholamine response between the two groups. 123I-MIBG scintigraphy showed that the heart/mediastinum ratio of MIBG uptake was significantly lower, and scintigraphic defect was greater in group B than in group A. There were no significant differences in the washout rate between the two groups. These results suggested that the lack of adrenergic symptoms during hypoglycemia may be associated with cardiac sympathetic nervous dysfunction in insulin-treated NIDDM patients, and this dysfunction is mainly due to cardiac sympathetic denervation.
Internal Medicine 03/1996; 35(2):94-9. · 0.97 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: A case of native valve endocarditis due to Acinetobacter calcoaceticus in a patient with dental caries is presented. The aortic, mitral and tricuspid valves were affected and showed vegetation by echocardiography in the affected valves. In spite of a good response to antibiotic therapy, multiple embolisms resulted in the patient's death. An autopsy confirmed the vegetation, which was calcified and contained no bacteria.
Japanese Circulation Journal 01/1991; 54(12):1563-5.
[Show abstract][Hide abstract] ABSTRACT: A 65-year-old man with idiopathic myocarditis is described. He was admitted with symptoms of acute heart failure. Examination revealed left ventricular hypokinesis. Results of an endomyocardial biopsy showed "resolving myocarditis". Immunological studies of the peripheral blood showed a low helper-suppressor (CD4/CD8) ratio on admission and depressed natural killer (NK) cell activity coincident with the onset of myocarditis. We considered that this immunological imbalance may have occurred during the progression of myocarditis to dilated cardiomyopathy.
Japanese Heart Journal 10/1990; 31(5):741-4. · 0.40 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Fifty-eight patients with symptomatic congestive heart failure were examined for T-lymphocyte subsets in the peripheral blood using two-color laser flow cytometry as a noninvasive diagnostic procedure. The final diagnosis established by catheterization and endomyocardial biopsy were dilated cardiomyopathy (DCM, n = 24), myocarditis (MC) by the Dallas criteria (n = 12), and coronary heart disease (CHD, n = 16). The CD8+CD11- (cytotoxic T) subset was significantly low in patients with DCM (13.9 +/- 4.4 vs. controls, p less than 0.05) in comparison with MC (20.7 +/- 10.9) and CHD (22.3 +/- 5.9). Moreover, the CD4+2H4+ (suppressor/inducer T) subsets were higher in patients with DCM (27.3 +/- 6.9 vs. controls, p less than 0.01) than in those with MC (17.3 +/- 7.8) and CHD (15.6 +/- 7.9). The CD4/CD8 and CD4+2H4+/CD8+CD11- ratio were examined and compared with those of normal controls (NC n = 16). The CD4+2H4+/CD8+CD11- ratio was clearly higher in patients with DCM (2.2 +/- 0.9 vs. controls, p less than 0001) than in those with MC (1.1 +/- 0.6) CHD (0.9 +/- 0.7). A CD4+2H4+/CD8+CD11- ratio of greater than 1.6 was considered to facilitate diagnosis of dilated cardiomyopathy with 79% sensitivity and 70% specificity. There was no significant increase in the ratios between MC and CHD. However, the proportion of the CD8+Leu7+ (natural suppressor) subset of circulating T lymphocytes in patients with MC was statistically higher (19.1 +/- 6.3% vs. controls, p less than 0.05) than in DCM or CHD. An elevated ratio of CD4+2H4+/CD8+CD11- among peripheral blood lymphocytes may thus be a useful marker for differential diagnosis of dilated chronic cardiomyopathy from myocarditis and coronary heart disease.
[Show abstract][Hide abstract] ABSTRACT: A 44 year-old woman died after a history of chronic right heart failure for 25 years. Roentgenological studies showed marked pulmonary aneurysm, and hemodynamics at rest revealed severe pulmonary hypertension. The diagnosis of chronic pulmonary thromboembolism was made by MRI which identified a high intensity mass in the dilated right pulmonary artery. At autopsy, a large organized thrombus adhered to the atherosclerotic pulmonary artery, and pulmonary thromboembolism or hemorrhagic infarction were identified. Microscopic examination revealed intimal and medial proliferation in small arteries and plexiform lesions. In addition, hypoplasty of the portal vein and portacaval shunt, which were thought to be congenital, were present. It has been recognized that the portacaval shunt can be attributed to pulmonary hypertension. In this case it was considered that the main cause of pulmonary thrombus formation was both pulmonary atherosclerosis and pulmonary artery aneurysm caused by prolonged pulmonary hypertension. This is the first case of chronic pulmonary thromboembolism associated with congenital hypoplasty of portal vein.
[Show abstract][Hide abstract] ABSTRACT: A pair of 37-year-old identical twins with diabetes mellitus are described. One of the brothers was admitted for heart failure without pain, and autonomic neuropathy was found. The clinical diagnosis was inferior myocardial infarction with anteroseptal healed myocardial infarction. Cardiac catheterization revealed triple coronary vessel involvement. The diagnosis was confirmed at autopsy after sudden death. The other brother was also examined by cardiac catheterization, which revealed total right coronary occlusion and hypokinesis of the wall. There had been no previous pain nor upper body discomfort until that time in either twin. Thus, genetic factors should possibly be considered in the genesis of asymptomatic or silent myocardial infarction.