Marked Suppression of T Cells by a Benzothiophene Derivative in Patients with Human T-Lymphotropic Virus Type I-Associated Myelopathy/Tropical Spastic Paraparesis

In a search for new anti-autoimmune agents that selectively suppress activation of autoreactive T cells, one such agent, 5-methyl-3-(1-methylethoxy)benzo[b]thiophene-2-carboxamide (CI-959-A), was found to be effective. This compound, which is known to suppress tumor necrosis factor alpha (TNF-α)-induced CD54 expression, inhibited the primary proliferative response of the T cell to antigen (Ag)-presenting cells (APCs) including allogenic dendritic cells (DCs), autologous Epstein-Barr virus-infected B cells, and human T lymphotropic virus type I (HTLV-I)-infected T cells. Autoreactive T cells from patients with HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP) spontaneously proliferate in vitro, and their activation is reported to be associated with CD54 expression. The spontaneous proliferation of T cells from patients with HAM/TSP was entirely blocked by CI-959-A. However, in this study, the T-cell proliferation in 15 patients with HAM/TSP was found to depend more extensively on major histocompatibility complex (MHC) class II and CD86 than on CD54 Ags. Since most important APCs for the development of HAM/TSP are DCs and HTLV-I-infected T cells, the effect of CI-959-A on DC generation and on the expression of surface molecules on activated T cells is examined. CI-959-A suppressed recombinant granulocyte-macrophage colony stimulating factor (GM-CSF)- and recombinant interleukin-4-dependent differentiation of DCs from monocytes and inhibited the expression of CD54 and, more extensively, MHC class II and CD86 Ags. CI-959-A showed little toxicity toward lymphoma or HTLV-I-infected T-cell lines or toward monocytes and cultured DCs. These results suggest that CI-959-A might be a potent anti-HAM/TSP agent.Human T lymphotropic virus type I (HTLV-I)-associated myelopathy/tropical spastic paraparesis (HAM/TSP) is thought to be an autoimmune disease induced by HTLV-I infection (8, 9, 24). The T lymphocytes obtained from patients with HAM/TSP patients produce interleukin-2 (IL-2) in vivo and proliferate spontaneously in vitro without any additional stimuli or cytokines (35). This spontaneous proliferation of T lymphocytes (SPL) depends on the interaction of T cells with antigen (Ag)-presenting cells (APCs) such as dendritic cells (DCs) (17, 25) and HTLV-I-infected CD4⁺ T cells (15, 32). The DCs localized in the blood and nonlymphoid organs are considered to be functionally immature, in that they are optimized for the uptake and processing of Ag but not for the initiation of primary T-cell responses. However, after the uptake of Ag and exposure to inflammatory agents including tumor necrosis factor alpha (TNF-α) and IL-1, the DCs undergo a process of maturation and gain the ability to present Ag to T cells for their priming (22, 26). In addition to DCs, HTLV-I-infected CD4⁺ T cells directly stimulate autologous CD4⁺ T cells in a major histocompatibility complex (MHC) class II- and CD86 molecule-dependent fashion (32). Among the T cells stimulated with these APCs, some might cross-react with self Ags and closely associate with the development of HAM/TSP.We have been searching for compounds that inhibit the cellular interaction between APCs and T cells to suppress the activation of autoreactive and Ag-specific T cells. The molecules associated with the APC-T cell interaction may provide an effective target for therapy for autoimmune diseases. Binding of APCs and T cells is initiated by contact of adhesion molecules, such as CD54 and CD11a/CD18, expressed on both cells, and induction of sustained proliferation of T cells requires two independent signals provided by APCs: a T-cell receptor-mediated Ag-specific signal and a signal mediated by costimulatory molecules (CSMs) (10, 20) including CD86 and CD58 Ags (1, 11, 31). Blocking of their tight binding through adhesion molecules or interaction of the CSMs with CSM ligands effectively suppressed the abnormal expansion of disease-associated T cells in vivo and in vitro (19, 30, 32) and sometimes effectively induced a long-term unresponsiveness of T cells to recall stimuli.5-Methyl-3-(1-methylethoxy)benzo[b]thiophene-2-carbox-amide (CI-959-A) is known to inhibit CD54 expression, and its derivative is reported to inhibit casein kinase II (4). In the present study, we found that CI-959-A markedly suppressed SPL in patients with HAM/TSP. Furthermore, the compound suppressed the primary T-cell proliferative response to stimuli provided by various APCs, the differentiation of immature DCs from monocytes and their subsequent maturation, and the induction of expression of MHC class II, CD54, and CD86 Ags on activated CD4⁺ T cells.     

In vitro schedule-dependent interaction between paclitaxel and SN-38 (the active metabolite of irinotecan) in human carcinoma cell lines

Paclitaxel and irinotecan are important new anticancer agents. The combination of these two agents has been considered for use against a variety of advanced solid tumors. Since the schedule-dependent effects of this combination may be crucial to its use, we studied the interaction of paclitaxel and SN-38 (the active metabolite of irinotecan) in various schedules in four human cancer cell lines in culture. Cell growth inhibition after 5 days was determined using an MTT assay. The effects of drug combinations at the IC₈₀ level were analyzed by the isobologram method. Simultaneous exposure to paclitaxel and SN-38 for 24 h produced antagonistic (subadditive and protective) effects in the human lung cancer cell line A549, the breast cancer cell line MCF7, and the colon cancer cell line WiDr, and produced additive effects in the ovarian cancer cell line PA1. Sequential exposure to paclitaxel for 24 h followed by SN-38 for 24 h, and the reverse sequence, produced additive effects in all four cell lines. These findings suggest that sequential administration, not simultaneous administration, may be the appropriate schedule for the therapeutic combination of paclitaxel and irinotecan. Continued preclinical and clinical studies should provide further insights and assist in determining the optimal schedule for this combination in clinical use. Received: 25 February 1997 / Accepted: 6 November 1997     

Telmisartan treatment decreases visceral fat accumulation and improves serum levels of adiponectin and vascular inflammation markers in Japanese hypertensive patients.

Hypertension contributes to the occurrence and progression of cardiovascular diseases. The angiotensin II type 1 receptor blocker telmisartan is reported to activate the peroxisome proliferator-activated receptor gamma and improve insulin sensitivity. We investigated the effects of telmisartan treatment on visceral fat, serum adiponectin and vascular inflammation markers in Japanese hypertensive patients. This was an open-label, non-controlled study. Twenty-eight essential hypertensive patients (22 men and 6 women; age 60.6+/-1.9 years; body mass index [BMI] 25.5+/-0.6 kg/m(2)) participated. Fat area was assessed with computerized tomography. All the subjects were started on telmisartan 40 mg/day, which was increased to 80 mg/day to achieve the blood pressure target of less than 130/80 mmHg. We assessed the visceral and subcutaneous fat areas, serum adiponectin levels, and vascular inflammation markers at baseline and 24 weeks of telmisartan treatment. There were significant reductions in visceral fat area (from 103.1+/-7.9 to 93.3+/-8.4 cm(2), p<0.01) and pulse wave velocity (from 1,706+/-52 to 1,587+/-51 cm/s, p<0.01) at 24 weeks. In contrast, significant increases in serum high-density lipoprotein cholesterol (from 5.06+/-0.15 to 5.32+/-0.13 mmol/L, p<0.05) and adiponectin levels (from 8.27+/-0.76 to 9.13+/-0.81 microg/mL, p<0.05) were observed. Also, there were reductions in the interleukin-6 level (from 2.26+/-0.27 to 1.60+/-0.14 pg/mL, p<0.01). We also conducted these investigations in male subjects alone and similar findings were obtained for all of these parameters. In conclusion, telmisartan treatment was associated with an improvement of vascular inflammation, reductions in visceral fat and increases in serum adiponectin.     

Taurine in the mammalian cerebellum: Demonstration by autoradiography with [3H]taurine and immunocytochemistry with antibodies against the taurine-synthesizing enzyme, cysteine-sulfinic acid decarboxylase

Taurine neurons and their dendrites and axons were visualized in the mammalian cerebellum by autoradiography, after in vivo injections of [³H]taurine directly into the cerebellar cortex or deep cerebellar nuclei, and by immunocytochemistry at the light- and electron-microscope levels with antibodies against cysteine-sulfinic acid decarboxylase (CSADCase; L-cysteine-sulfinate carboxylyase, EC 4.1.1.29). Uptake and sequestration of [³H]taurine labeled numerous Purkinje cell somata, primary dendrites, and axons; many granule cell somata, dendrites, and parallel fibers; stellate, basket, and Golgi cells; the larger neurons in all deep cerebellar nuclei; the largest neurons in the lateral vestibular nucleus; and, more rarely, Purkinje cell axonal terminals in the neuropil. The label at all sites was diminished by preinjection into the cerebellum of hypotaurine, p-chloromercuriphenylsulfonic acid, or β-alanine, and was virtually eliminated by strychnine. Immunocytochemical labeling with polyclonal antibodies directed against CSADCase, the enzyme responsible for the synthesis of hypotaurine from cysteine sulfinic acid and taurine from cysteic acid, had a similar distribution. In electron micrographs, immunoreactivity within Purkinje cell somata and dendrites was localized to the Golgi apparatus, the inner plasma membrane, and condensed nonmembranous foci (120 nm in diameter) marked by clumps of peroxidase reaction product. Large Nissl bodies were usually not CSADCase immunoreactive. Numerous immunoreactive granule cells, dendrites, and parallel fibers were recognized. Pretreatment of the animals with colchicine increased the intensity of CSADCase immunoreactivity but did not change the number or distribution of labeled cells. These experiments indicate that taurine is synthesized and involved in a specific uptake process by cerebellar neurons. Neuroglial cells do not synthesize taurine but some neuroglia take up [³H]taurine. These findings call for a reexamination of the physiological function of taurine in the cerebellum. A hypothesis is proposed that taurine may be involved in the regulation of calcium, in dendritic spike generation, and in the inhibition of impulse propagation in major Purkinje cell dendrites.