Immunosuppressive effect of FTY 720 on autoimmune diabetes models.
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Biomedical subjects
Publications and source records attributed to H Amemiya.
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There have been controversies concerning the optimal target sites and approaches in radiofrequency catheter ablation of common atrial flutter. We attempted high energy radiofrequency catheter ablation targeting the isthmus between the inferior vena cava and tricuspid valve annulus (IVC-TV isthmus) with a super long (8 mm) tip electrode, and compared the efficacy of this anatomical approach with the electrophysiological approach targeting the posteroseptal right atrium posterior to the coronary sinus using a standard 4-mm tip electrode. Atrial flutter was successfully ablated in 12 of 12 patients (100%) without recurrence with the anatomical approach, while, in 7 of 9 patients (64%) with 2 recurrences with the electrophysiological approach. In comparison of ablation data between the anatomical and electrophysiological approaches, there were significant differences in the mean number of application pulses (anatomical vs electrophysiological: 2.3 +/- 0.8 vs 9.9 +/- 6.4, P < 0.01), applied wattage (39 +/- 12 W vs 24 +/- 6 W, P < 0.01), applied energy per application (1,986 +/- 426 J vs 659 +/- 323 J, P < 0.01), fluoroscopic time (26 +/- 11 min vs 74 +/- 30 minutes, P < 0.01), and procedure time (59 +/- 8 min vs 181 +/- 53 min, P < 0.01). In conclusion, the anatomical approach is superior to the electrophysiological one with respect to procedure and radiation time, and linear ablation at the IVC-TV isthmus with an 8-mm tip electrode and high energy application is highly effective and safe.
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The in vitro treatment of lpr thymocytes with FTY720 resulted in a dose-dependent reduction in cell viability due to apoptosis. In order to study the effect of FTY720 in vivo, lpr mice received an oral daily dose of 1 mg/kg FTY720 for 14 days, beginning at 16 weeks of age which was when the animals were developing massive lymphoadenopathy. Compared with untreated lpr mice, FTY720-treated lpr mice had significantly prolonged lives. At 24 weeks of age, treated mice demonstrated markedly reduced weights of the spleen and lymph nodes, and the proportion of CD3+ B220+ and CD4- CD8- cells in the thymus, spleen and lymph nodes decreased markedly. In addition, in these mice the percentage of CD4+ CD8+ and CD3- B220- cells in the thymus and the percentage of CD4+ CD8-, CD4- CD8+, CD3+ B220- and CD3- B220+ cells in the spleen returned to almost the normal values observed in wild-type mice. Histological observation 1 day after the final administration of FTY720 revealed a remarkable infiltration of neutrophils in the lymphoid organs. Apoptotic cells were detected in all the lymphoid organs using in situ DNA nick-end labelling. Electron microscopy showed that the apoptotic cells were ingested by phagocytes. FTY720 therapy is thus highly effective in Fas-mutant animals with abnormally expanding lymphocytes.
We previously reported that FTY720 is an efficient inducer of apoptosis in lymphocytes and cultured cell lines. In the present study, HL-60 human promyerocytoma cells also induced apoptosis through in vitro treatment with the drug, demonstrating extensive DNA fragmentation 6 hr after incubation. The major target of FTY720 was the common signalling pathway of apoptosis, since a rapid (< 1 min) increase in the intracellular Ca2+ concentration ([Ca2+]i) was found in the cells treated with the drug. Calcium chelation in the culture medium with EGTA did not affect the [Ca2+]i mobilization. A phospholipase C inhibitor, U73122, inhibited the increase in [Ca2+]i as well as the fragmentation of the nuclear DNA, whereas U73343, a non-effective analogue of U73122, had little effect. These results suggest that FTY720-induced apoptosis is mediated through an activation of phospholipase C and the subsequent release of Ca2+ from intracellular calcium pools. In addition, the treatment of HL-60 with pertussis toxin (PTX) did not inhibit Ca2+ mobilization or apoptosis, suggesting that the activation of phospholipase C is independent of PTX-sensitive G-proteins.
The electrographic features of successful sites of radiofrequency catheter ablation were analyzed in 33 cases of posteroseptal accessory pathways and compared with those from 155 cases of free wall accessory pathways. The atrioventricular intervals in the posteroseptal cases were significantly longer than in the free wall cases (posteroseptal vs left and right free wall; 38 vs 33 and 26 msec, respectively; p < 0.05), and the incidences of continuous electrograms (42 vs 63 and 79%; p < 0.01) and PQS-pattern unipolar electrograms (50 vs 76 and 78%; p < 0.05) were significantly lower in the posteroseptal cases. The V-delta intervals in the posteroseptal cases were significantly longer than in the left free wall cases (17 vs 13 msec; p < 0.05), but shorter than in the right free wall cases (17 vs 23 msec; p < 0.05). No statistically significant difference in the incidence of Kent potentials among the 3 groups was observed. In radiofrequency ablation of posteroseptal pathways, the length of the atrioventricular interval and the incidences of continuous electrograms and PQS-pattern unipolar electrograms may be unsatisfactory even at the appropriate target site, but the V-delta interval and Kent potential are good indicators of suitable target sites.
Transplant renal arteriopathy (TRA) is a major obstacle to the long-term survival of human renal allografts. Tenascin (TN) is an extracellular matrix glycoprotein associated with cell growth and differentiation. We investigated TN expression in intrarenal arteries with TRA, in association with cellular components, with phenotypic expression of smooth muscle cells (SMC) and with fibronectin expression. Ten renal allografts that had been removed due to rejection were available. Monoclonal antibodies against SMC, macrophages, T cells, B cells, fibronectin, and TN were used. In the early stages, medial SMC showed a de-differentiated phenotype and the neointima consisted largely of T cells and macrophages. At these stages, increased expression of TN was observed in the media. In later stages, the neointima consisted almost entirely of SMC of a differentiated phenotype and no TN expression was found. Up-regulation of TN may play a role in the migration and phenotypic modulation of SMC at an early stage of TRA in humans.
To evaluate whether a high level of lipoprotein(a) (Lp(a)) is a risk for the development of coronary heart disease (CHD), 94 Japanese patients and 64 age-matched Japanese controls, diagnosed after coronary angiography (CAG), were analyzed with special reference to the relations between the degree of atherosclerosis, Lp(a) levels and the apolipoprotein(a) (apo(a)) genotypes. the degree of atherosclerosis was evaluated based on CAG findings in the following three ways: the number of diseased vessels, the Gensini score, and the presence or absence of vascular ulcers and/or irregular outlines of coronary stenotic lesions. Apo(a) protein sizes and the pentanucleotide (TTTTA) repeat polymorphism in the 5' control region of the apo(a) gene were analyzed. Multivariate predictors for the number of diseased vessels were, in decreased order of significance, plasma Lp(a) levels, history of smoking, hypertension, diabetes mellitus, and body mass index (BMI). Independent factors associated with the Gensini score were Lp(a) levels, BMI, hypertension, and diabetes mellitus. A negative association of Lp(a) levels with apo(a) protein sizes, and higher Lp(a) levels in those homozygous for an allele with 8 8 (TTTTA)-repeats, was found in both the controls and patients. In decreasing order of significance, apo(a) protein sizes, the degree of atherosclerosis, the genotype of the pentanucleotide repeat, and gender were independent predictors of Lp(a) levels in stepwise regression models. Apo(a) protein sizes were a significant predictor, and the genotype homozygous for the 8 (TTTTA)-repeats was a possible predictor, for the degree of atherosclerosis in CHD. These findings support the notion that a high Lp(a) level is a risk for the development of atherosclerosis in CHD.
A novel immunosuppressant, FTY720, induces a rapid and marked decrease of peripheral lymphocytes, and prolong allograft survival in rats. Its mechanism of action is mediated by apoptotic cell death. In this study, we determined the time-related changes in the numbers of total lymphocytes, and the ratios of lymphocyte subpopulations in peripheral blood and lymphomyeloid organs in rats after the single oral administration of FTY720 (10 mg/kg), comparing with the effects of cyclophosphamide (80 mg/kg, ip). Total number of peripheral lymphocytes decreased significantly 3 h after the administration of the drug, while that of polymorphonuclear cells increased. T-cells were markedly decreased in number and reached a minimum of 2.3% of the control 3 days after the treatment, while B-cells reached 19.7%. T-cells decreased in spleen and liver but there was no notable change in thymus, lymph nodes, and bone marrow. The susceptibility of the cells against the drug was variant based on the type and the source of cells in vitro. Polymorphonuclear cells were the most resistant and lymph node cells the most sensitive to FTY720 after 3 h incubation with different concentration of the drug (1, 10,100 mumol/l). When incubated with 10 mumol/l of FTY720, B-cells were significantly higher in viability than the whole T- or CD4-cells. These results demonstrated that FTY720 induces cell death selectively in mature T-lymphocytes, especially CD4-lymphocytes, in peripheral blood without the depression of bone marrow.
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15-Deoxyspargualin (DSG) is a synthetic analogue of spergualin isolated from the culture filtrate of Bacillus laterosporus. It shows a strong immunosuppressive effect by antiproliferating action inhibiting the IL-2-stimulated maturation of T cells from the G0/G1 phases to the S and G2/M phases. Hsc70, a constitutive member of Hsp70 was identified as the immunophilin of DSG by Nadlar et al. In allogeneic transplantation of rat heart and dog kidney, DSG was definitely proved to prevent rejection and to rescue ongoing rejection. Our multicenter clinical trials on DSG showed that the percent efficacy to reverse acute rejection was 70-80%, and 600 days graft survival was 90% in the cases effectively treated with DSG at acute rejections. The combined use of DSG with methylprednisolone to treat acute rejection and the use of DSG to treat rejections in chronic phases were reported as the beneficial uses of DSG.
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