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Hitomi Nagayama,
Kazufumi Matsumoto,
Naoyuki Isoo,
Hideki Ohno,
Naoyuki Takahashi, Takashi Nakaoka,
Masaru Shinozaki,
Makoto Watanabe,
Yusuke Inoue,
Fumitaka Nagamura,
Naoki Oyaizu,
Naohide Yamashita
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ABSTRACT: Most pancreatic cancer patients are diagnosed at the advanced stages, and no therapy is superior to gemcitabine alone. To
confirm the feasibility and efficacy of a novel clinical intervention using tumor vessel-specific anti-angiogenic peptide
vaccination, we conducted a clinical phase I/II trial using HLA-A*2402/A*0201-restricted vascular endothelial growth factor
receptor type 1 (VEGFR1)-derived peptide vaccination in combination with gemcitabine for advanced pancreatic cancer (http://www.clinical-trials.gov; NCT00683358 and NCT00683085). Four of the enrolled patients (n=2 for HLA-A*2402 and n=2 for HLA-A*0201 protocol, respectively), defined as having progressive disease according to the Response Evaluation Criteria
in Solid Tumors version 1.0 (RECIST v.1.0), failed to respond to the therapy. Another two patients enrolled in HLA-A*2402
protocol dropped out of the study due to rapid disease progression. Grade 2–3 hematologic toxicities were observed in all
cases, but the treatment was well tolerated with minimal systemic adverse events. One case in HLA-A*2402 protocol and another
case in HLA-A*0201 protocol suffered complicated gastrointestinal (GI) bleeding during vaccination. The causal relationship
between GI bleeding and VEGFR1-peptide vaccination is unclear according to the pathologic examination. These studies terminated
prematurely because of the advanced stage of the disease in the enrolled patients on entry to the study. Despite GI bleeding,
peptide vaccination provides a feasible treatment option for many advanced pancreatic cancer patients.
KeywordsPancreatic cancer-Peptide vaccination-VEGFR1-Gemcitabine-GI bleeding
Clinical Journal of Gastroenterology 04/2012; 3(6):307-317.
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ABSTRACT: Cord blood (CB) transplantation has advantages in terms of incidence and severity of acute graft-versus-host disease (GVHD), while it has disadvantages in terms of infection. Our aim is to elucidate the molecular mechanism underlying the immune response of CB-derived cells during acute GVHD and infection following CB transplantation.
We examined expression of 69 immune-relating microRNAs in CD4(+), CD8(+), and CD14(+) cells of CB and adult peripheral blood (APB) upon interferon-γ or lipopolysaccharide (LPS) stimulation.
Under basal condition, 20 microRNAs showed differential expression between CB and APB. Compared to APB counterparts, six microRNAs (miR-21, miR-22, miR-29a, miR-29b, miR-29c, and let-7c) were underexpressed in at least two cell lineages of CB, while five microRNAs (miR-15b, miR-181a, miR-181c, miR-363, and miR-424) were overexpressed in CD4(+) and CD8(+) CB cells. Upon interferon-γ stimulation, seven microRNAs (miR-29a, miR-29b, miR-34c-5p, miR-132, miR-146a, miR-146b-5p, and miR-155) changed in expression mainly in CD14(+) CB cells, while only two microRNAs (miR-18a and miR-155) changed in expression in CD14(+) CB cells upon LPS stimulation. These results suggest that the mechanisms regulating the expression of such immune-relating microRNAs in CD14(+) CB cells are much more sensitive to proinflammatory stimuli than those in APB CD14(+) cells, which might be related to the poor immunoreactivity of CD14(+) CB cells.
Our results suggest essential roles of specific microRNAs in regulating immune function of CB cells, providing insight into the underlying molecular mechanism.
European Journal Of Haematology 09/2011; 88(1):31-8. · 2.61 Impact Factor
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ABSTRACT: Key role of angiogenesis in tumor growth and metastasis based on accumulating evidence and recent progress of immunotherapy have led us to investigate vaccine therapy targeting tumor angiogenesis.
C57BL/6J mice were vaccinated with a syngeneic endothelial cell line Tpit/E by subcutaneous injection once a week. Prior to ninth vaccination, the mice were challenged with B16/F10 melanoma cells by subcutaneous inoculation on the back for the tumor growth model or by tail venous injection for the lung metastasis model. Development of subcutaneous tumor and lung metastasis was monitored by computed tomography scanning, which enabled accurate evaluation with the minimized sacrifice of mice.
Vaccination with Tpit/E cells inhibited subcutaneous tumor growth and appearance of lung metastasis compared to control. Survival period was elongated in the Tpit/E vaccination in both of the two models. We also obtained hybridomas secreting specific antibodies to Tpit/E cells from a mouse vaccinated with the cells, indicating that specific immune response to the syngeneic endothelial cells was elicited.
These results suggest that vaccination with an autologous endothelial cell line may be effective against melanoma.
Journal of Experimental & Clinical Cancer Research 02/2009; 28:13. · 2.15 Impact Factor
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ABSTRACT: Dnm3os, a gene that is transcribed into a non-coding RNA (ncRNA), contains three micro RNAs (miRNAs), miR-199a, miR-199a*, and miR-214, whose functions remain unknown in mammals. In this study, we introduced the lacZ gene into the Dnm3os locus to recapitulate its expression pattern and disrupt its function. Dnm3os(+/lacZ) heterozygous embryos showed beta-galactosidase activity, which reflected the authentic expression pattern of Dnm3os RNA. Most of the Dnm3os(lacZ/lacZ) homozygous pups died within one month of birth. After birth, Dnm3os(lacZ/lacZ) mice exhibited several skeletal abnormalities, including craniofacial hypoplasia, defects in dorsal neural arches and spinous processes of the vertebrae, and osteopenia. Importantly, the expression of miR-199a, miR-199a*, and miR-214 was significantly down-regulated in Dnm3os(lacZ/lacZ) embryos, supporting the assumption that Dnm3os serves as a precursor of these three miRNAs. Thus, Dnm3os has emerged as an miRNA-encoding gene that is indispensable for normal skeletal development and body growth in mammals.
Developmental Dynamics 12/2008; 237(12):3738-48. · 2.54 Impact Factor
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Koichiro Kuwabara,
Toshihide Nishishita,
Mariko Morishita,
Naoki Oyaizu,
Shunichi Yamashita,
Takashi Kanematsu,
Takao Obara,
Yoshikazu Mimura,
Yusuke Inoue,
Michio Kaminishi, [......],
Akira Miyauchi,
Shiro Noguchi,
Kaoru Uchimaru,
Eiji Akagawa,
Nobukazu Watanabe,
Tsuneo A Takahashi,
Kaori Sato,
Takeshi Inazawa, Takashi Nakaoka,
Naohide Yamashita
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ABSTRACT: We assessed the feasibility and efficacy of dendritic cell (DC) therapy for advanced thyroid papillary and follicular cancer.
Six Japanese patients (2 men and 4 women; aged 46-72 years, mean 60 years), who were diagnosed as advanced thyroid cancer with refractory distant metastases (papillary, n=5; follicular, n=1), were enrolled. Patients were first vaccinated weekly for 4 weeks with 10(7) autologous tumor lysate-pulsed monocyte-derived mature DCs followed by fortnightly vaccinations for 8 weeks (total=8 vaccinations). Lowdose (350 KIU) interleukin-2 was also administered for 3 days at each vaccination. Clinical response, adverse effects, delayed-type hypersensitivity skin testing (DTH), and IFN-( ) production by peripheral CD3(+) lymphocytes were evaluated.
Of the 6 patients, disease was assessed as stable in 2 and as progressive in 4. No adverse events were observed. Results of DTH and IFN-( ) production in peripheral lymphocytes did not correlate to the clinical response.
DC immunotherapy could be administered to patients with thyroid papillary or follicular cancer without substantial side effects.
Thyroid 02/2007; 17(1):53-8. · 4.79 Impact Factor
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ABSTRACT: The aims of this study were to investigate the applicability of Fourier fitting in the magnetic resonance (MR) evaluation of left ventricular (LV) function and to determine the optimal number of harmonics for fitting. Cine cardiac MR imaging was performed in 10 subjects, and an LV time-volume curve was generated. Fourier fitting was applied to the original curve using 1-10 harmonics, and the qualities of the time-volume curve and first-derivative curve were evaluated. LV functional parameters were calculated from curves generated with and without fitting. The quality of the original time-volume curve was good, and Fourier fitting had no substantial effect on functional parameters obtained directly from the time-volume curve such as ejection fraction. The first-derivative curve generated without fitting showed substantial artificial fluctuation. The application of Fourier fitting depressed the fluctuation and tended to decrease estimates of peak ejection rate and peak filling rate. Five or six harmonics appeared to be appropriate for obtaining a high-quality first-derivative curve. In conclusion, Fourier fitting was indicated to aid in reducing the artificial fluctuation of the first-derivative curve generated from cine cardiac MR imaging and to contribute to the evaluation of functional parameters derived from the first-derivative curve.
Magnetic Resonance Imaging 01/2007; 24(10):1333-9. · 1.99 Impact Factor
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Akira Tomonari,
Satoshi Takahashi,
Jun Ooi, Takashi Nakaoka,
Kashiya Takasugi,
Michihiro Uchiyama,
Nobuhiro Tsukada,
Takaaki Konuma,
Tohru Iseki,
Arinobu Tojo,
Shigetaka Asano
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ABSTRACT: The cytotoxic effect of cytarabine (Ara-C) on myeloid leukemic cells is enhanced by concomitant use of granulocyte colony-stimulating factor (G-CSF) in vitro. The feasibility of a conditioning regimen consisting of G-CSF-combined 24 g/m2 Ara-C, 90 mg/m2 fludarabine, and 12 Gy total body irradiation was studied for five patients with acute myelogenous leukemia in cord blood transplantation (CBT). Graft vs. host disease (GVHD) prophylaxis consisted of cyclosporine and methotrexate. After the conditioning regimen, 2.48 x 10(7)/kg (2.28-3.53) of cord blood nucleated cells was infused. Neutrophil counts consistently >0.5 x 10(9)/L was achieved 24 d (22-32) after CBT. Grade I stomatitis and gastrointestinal toxicities occurred in all patients. Grades I and II acute GVHD occurred in one and four patients, respectively, which resolved without steroid therapy. Sepsis and aspergillosis occurred in two and one patients, respectively. All patients were alive without leukemia relapse at a follow up of 15 months (12-43) after CBT. This conditioning regimen could avoid the toxicities of high-dose cyclophosphamide but might enhance the cytotoxic effect of Ara-C. Large-scale studies will be needed to determine the efficacy and safety of the conditioning regimen in CBT.
European Journal Of Haematology 08/2006; 77(1):46-50. · 2.61 Impact Factor
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Kenta Yoshiura, Takashi Nakaoka,
Toshihide Nishishita,
Katsuaki Sato,
Akifumi Yamamoto,
Shinji Shimada,
Toshiaki Saida,
Yutaka Kawakami,
Tsuneo A Takahashi,
Hiroyuki Fukuda,
Shinobu Imajoh-Ohmi,
Naoki Oyaizu,
Naohide Yamashita
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ABSTRACT: Tumor-associated antigens are promising candidates as target molecules for immunotherapy and a wide variety of tumor-associated antigens have been discovered through the presence of serum antibodies in cancer patients. We previously conducted dendritic cell therapy on 10 malignant melanoma patients and shrinkage or disappearance of metastatic tumors with massive necrosis occurred in two patients. In this study, we found a 29-kDa protein against which antibody was elicited by dendritic cell therapy in one of the two patients. Matrix-assisted laser desorption ionization-time of flight/mass spectrometry analysis of the protein isolated by two-dimensional electrophoresis combined with Western blots revealed that the 29-kDa protein was carbonic anhydrase II (CA-II). Immunohistochemistry of the tumors and normal tissues showed that CA-II was expressed in the tumor vessel but not in normal vessel endothelium. CA-II expression in tumor endothelium was observed as well in other cancers including esophageal, renal, and lung cancers. In an in vitro angiogenesis model, CA-II expression of normal human vein endothelial cells was significantly up-regulated when cells were cultured in the acidic and hypoxic conditions indicative of a tumor environment. These findings suggest that CA-II is a tumor vessel endothelium-associated antigen in melanoma and other cancers, and elicitation of serum anti-CA-II antibody by dendritic cell therapy may be associated with good clinical outcome including tumor reduction.
Clinical Cancer Research 12/2005; 11(22):8201-7. · 7.74 Impact Factor
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ABSTRACT: The aim of this study was to investigate the effect of temporal resolution on the estimation of left ventricular (LV) function by cardiac magnetic resonance (MR) imaging using a steady-state free precession (SSFP) sequence. Left ventricular function was assessed by cine MR imaging using a segmented SSFP sequence in 10 healthy volunteers. Views per segment (VPS) were set at 8 and 20, resulting in high and low true temporal resolution, respectively. Irrespective of VPS, images were reconstructed at 40 cardiac phases, providing high apparent temporal resolution. Data were analyzed using 40, 20 and 10 phases to simulate different apparent temporal resolutions. Increasing the cardiac phases used for analysis slightly decreased mean end-systolic volume (ESV) and slightly increased mean ejection fraction (EF). No substantial difference in estimates of end-diastolic volume (EDV) was found between VPSs of 8 and 20. Imaging with a VPS of 20 yielded a larger ESV and smaller EF than imaging with a VPS of 8 when 40 phases were used. In conclusion, low true temporal resolution causes overestimation of ESV and underestimation of EF. Improvement of apparent temporal resolution mildly reduces but does not eliminate the errors caused by low true temporal resolution.
Magnetic Resonance Imaging 07/2005; 23(5):641-5. · 1.99 Impact Factor
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ABSTRACT: TRAIL preferentially induces apoptotic cell death in a wide variety of transformed cells, whereas it induces no apoptosis, but inhibits activation of Ag-specific T cells via blockade of cell cycle progression. Although accumulating results suggest that TRAIL is involved in the maintenance of immunological homeostasis under steady state conditions as well as in the initiation and progression of immunopathologies, the potential regulatory effect of TRAIL on immune responses and its therapeutic potential in immunological diseases remains unclear. We report in this study the potential usefulness of TRAIL-transduced dendritic cells (DCs) for the treatment of lethal acute graft-vs-host disease (GVHD) and leukemia relapse. DCs genetically modified to express TRAIL showed potent cytotoxicity against both alloreactive T cells and leukemic cells through the induction of apoptosis. In addition, treatment with genetically modified DCs expressing TRAIL of allogeneic BM transplants recipients with leukemia was effective for protection against acute GVHD and leukemia relapse. Thus, gene transfer of TRAIL to DCs is a novel modality for the treatment of acute GVHD and leukemia relapse by selective targeting of pathogenic T cells and leukemic cells.
The Journal of Immunology 05/2005; 174(7):4025-33. · 5.79 Impact Factor
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Toshihide Nishishita,
Kunie Ouchi,
Xiaohong Zhang,
Mariko Inoue,
Takeshi Inazawa,
Kenta Yoshiura,
Koichiro Kuwabara, Takashi Nakaoka,
Nobukazu Watanabe,
Koichi Igura,
Tsuneo A Takahashi,
Naohide Yamashita
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ABSTRACT: Recently several strategies to treat ischemic diseases have been proposed but the ideal way has to be determined. We explored whether human placenta-derived mesenchymal cells (hPDMCs) can be used for this purpose because placenta is very rich in vessels. First, production of human vascular endothelial growth factor (hVEGF) from hPDMCs was examined. The amount of hVEGF secreted by hPDMCs was similar to the amount produced by HeLa cells. hVEGF was barely detected in human umbilical vein endothelial cells (hUVECs) or human peripheral blood mononuclear cells. hVEGF secreted from hPDMCs stimulated the proliferation of hUVECs, indicating its biological activity. Transplantation of hPDMCs to the ischemic limbs of NOD/Shi-scid mice significantly improved the blood flow of the affected limbs. Blood vessel formation was more prominently observed in the limbs of treated mice as compared to the control mice. Real-time RT-PCR revealed that hPDMCs produced hVEGF for at least 7 days after transplantation. Thus, transplantation of hPDMCs could potentially be a promising treatment for human ischemic diseases.
Biochemical and Biophysical Research Communications 01/2005; 325(1):24-31. · 2.48 Impact Factor
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ABSTRACT: The current regimens of hormone replacement therapy for postmenopausal women, estrogen combined with progestogen, have failed to show beneficial effects for the prevention of atherosclerotic disease. Although the relatively higher dose of estrogen contained in those regimens exerted adverse effects, there are few data examining a lower dose of estrogen in an atherosclerosis model. Therefore, we investigated experimentally whether lower doses of estrogen could inhibit neointimal formation after balloon injury of the rat carotid artery. Ten-week-old Wistar rats were subjected to ovariectomy or sham-operation (n=7). Four days after ovariectomy, rats were implanted with an osmotic mini-pump containing 17-beta estradiol (0.2, 1, 2, 10 and 20 microg/kg/day; n=6, 4, 8, 6 and 5, respectively) or placebo (n=10). After 3 days of hormone therapy, balloon injury was performed in the left common carotid artery. Neointimal formation was histologically evaluated 2 weeks after injury. Cross-sectional intimal area and the ratio of intimal area to medial area were dose-dependently reduced by estrogen replacement compared with those in ovariectomized rats without estrogen replacement. The effects of estrogen replacement were identical to those of an angiotensin II type 1 receptor blocker, candesartan. Interestingly, the effect was significant even in rats receiving lower doses of estrogen, in which plasma estradiol concentrations were not increased and the hyperplastic response of the uterus was minimal. These results suggest the efficacy of low-dose estrogen therapy for the protection of atherosclerosis.
European Journal of Pharmacology 11/2004; 502(3):265-70. · 2.52 Impact Factor
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ABSTRACT: Proline-rich tyrosine kinase 2 (Pyk2), a member of the focal adhesion kinase family, is thought to act as a key component in vasculogenesis and angiogenesis. Therefore, we studied the effect of mutant Pyk2 expression on the migration and proliferation in endothelial cells (ECs). Two types of mutant Pyk2 were examined by adenovirus vectors AxCA-Pyk2K457A, expressing a kinase inactive mutant, and AxCA-Pyk2Y402F, expressing a tyrosine autophosphorylation site mutant, in addition to AxCA-Pyk2, expressing wild-type Pyk2. Migration of ECs infected with AxCA-Pyk2Y402F increased to a level similar to that of ECs infected with AxCA-Pyk2. The size of effect was dependent on the amount of applied adenoviruses within the range of 3-30 multiplicity of infection. In contrast, AxCA-Pyk2K457A infection did not show any significant effect on cell migration. Western blotting showed that both phosphorylation of Pyk2 Y(881) and association of p130(Cas) with Pyk2 were enhanced in ECs infected with AxCA-Pyk2Y402F as well as with AxCA-Pyk2, but not in ECs infected with AxCA-Pyk2K457A. Therefore, signaling mediated by Pyk2 Y(881) and p130(Cas) may be involved in the migration of ECs infected either with AxCA-Pyk2Y402F or with AxCA-Pyk2. In proliferation assay, AxCA-Pyk2 infection suppressed EC proliferation significantly; however, neither AxCA-Pyk2Y402F nor AxCA-Pyk2K457A showed such an inhibitory effect. Thus, the two Pyk2 mutants revealed that Pyk2 signaling differentially regulates cell migration and proliferation pathways.
Endocrinology 08/2004; 145(7):3324-30. · 4.46 Impact Factor
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Yuichi Ikeda,
Yasushi Imai,
Hidetoshi Kumagai,
Tetsuya Nosaka,
Yoshihiro Morikawa,
Tomoko Hisaoka,
Ichiro Manabe,
Koji Maemura, Takashi Nakaoka,
Takeshi Imamura,
Kohei Miyazono,
Issei Komuro,
Ryozo Nagai,
Toshio Kitamura
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ABSTRACT: Growth factors, cell-surface receptors, adhesion molecules, and extracellular matrix proteins play critical roles in vascular pathophysiology by affecting growth, migration, differentiation, and survival of vascular cells. In a search for secreted and cell-surface molecules expressed in the cardiovascular system, by using a retrovirus-mediated signal sequence trap method, we isolated a cell-surface protein named vasorin. Vasorin is a typical type I membrane protein, containing tandem arrays of a characteristic leucine-rich repeat motif, an epidermal growth factor-like motif, and a fibronectin type III-like motif at the extracellular domain. Expression analyses demonstrated that vasorin is predominantly expressed in vascular smooth muscle cells, and that its expression is developmentally regulated. To clarify biological functions of vasorin, we searched for its binding partners and found that vasorin directly binds to transforming growth factor (TGF)-beta and attenuates TGF-beta signaling in vitro. Vasorin expression was down-regulated during vessel repair after arterial injury, and reversal of vasorin down-regulation, by using adenovirus-mediated in vivo gene transfer, significantly diminished injury-induced vascular lesion formation, at least in part, by inhibiting TGF-beta signaling in vivo. These results suggest that down-regulation of vasorin expression contributes to neointimal formation after vascular injury and that vasorin modulates cellular responses to pathological stimuli in the vessel wall. Thus, vasorin is a potential therapeutic target for vascular fibroproliferative disorders.
Proceedings of the National Academy of Sciences 08/2004; 101(29):10732-7. · 9.68 Impact Factor
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Tokumitsu Watanabe,
Masahiro Akishita, Takashi Nakaoka,
Hong He,
Yukiko Miyahara,
Naohide Yamashita,
Youichiro Wada,
Hiroyuki Aburatani,
Masao Yoshizumi,
Koichi Kozaki,
Yasuyoshi Ouchi
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ABSTRACT: Estrogen has diverse effects on the vasculature, such as vasodilation, endothelial growth and inhibition of vascular smooth muscle cell (VSMC) proliferation and migration. However, little is known about the genes that are regulated by estrogen in the vascular wall. Wistar rats were ovariectomized or sham-operated (Sham group), and 2 weeks after the operation, were subjected to subcutaneous implantation of placebo pellets (OVX + V group) or estradiol pellets (OVX + E group). Endothelium-denuded aortic tissue was examined 2 weeks after implantation. By applying high-density oligonucleotide microarray analysis, the expression of approximately 7000 genes was analyzed. Among the genes with different expression levels between the OVX + E group and the OVX + V group, those that have been reported to be expressed in the vasculature or muscle tissue, were chosen. Finally, four genes, caveolin-1, two LIM proteins (enigma and SmLIM) and Id3a, were identified. Microarray as well as real-time polymerase chain reaction showed that the expression levels of these genes were significantly higher in the OVX + E group than in the OVX + V group. To clarify whether estrogen directly upregulates these genes in the vascular wall, Northern blot analysis was performed using cultured rat VSMC. Addition of 100 nmol/L estradiol for 24 hours increased the mRNA levels of all four genes. Although the precise mechanism remains unclear, regulation of these genes by estrogen might contribute to its effect on VSMC.
Life Sciences 08/2004; 75(10):1219-29. · 2.53 Impact Factor
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ABSTRACT: In our previous study, adrenomedullin (AM) overexpression could limit the arterial intimal hyperplasia induced by cuff injury in rats. However, it remains to be elucidated whether endogenous AM plays a role against vascular injury.
We used the AM knockout mice to investigate the effect of endogenous AM. Compared with wild-type (AM+/+) mice, heterozygous AM knockout (AM+/-) mice had the increased intimal thickening of the cuff-injured femoral artery, concomitantly with lesser AM staining. In AM+/- mice, cuff placement increased both the production of superoxide anions (O2-) measured by coelentarazine chemiluminescence and the immunostaining of p67phox and gp91phox, subunits of NAD(P)H oxidase in the adventitia, associated with the increment of CD45-positive leukocytes, suggesting that the stimulated formation of radical oxygen species accompanied chronic adventitial inflammation. Not only the AM gene transfection but also the treatment of NAD(P)H oxidase inhibitor apocynin and membrane-permeable superoxide dismutase mimetic tempol could limit cuff-induced intimal hyperplasia in AM+/- mice, associated with the inhibition of O2- formation in cuff-injured artery.
The overproduction of oxidative stress induced by the increased NAD(P)H oxidase activity might be involved in cuff-injured arterial intimal hyperplasia in AM+/- mice. Thus, it is suggested that endogenous AM possesses a protective action against the vascular response to injury, possibly through the inhibition of oxidative stress production.
Circulation 04/2004; 109(9):1147-53. · 14.74 Impact Factor
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ABSTRACT: The effect of 17 beta-estradiol (E2) on the proliferation of cardiac fibroblasts (CFs) remains controversial. This study investigated which subtype of estrogen receptor (ER), ER alpha or ER beta, mediated the effect of E2 on CF growth by the gain of function analysis using an adenovirus vector. One hundred nanomoles per liter of E2 attenuated DNA synthesis by up to 10%, and transactivated the estrogen-responsive element determined by luciferase assay in rat neonatal CFs. We constructed replication-deficient adenoviruses bearing the coding region of human ER alpha, ER beta, or the dominant-negative form of ER beta (designated AxCAER alpha, AxCAER beta, and AxCADNER beta, respectively). When CFs were infected with AxCAER alpha or AxCAER beta at multiplicity of infection of 20 or higher, DNA synthesis was decreased by 50% in response to E2 and the effect was abolished by co-infection with AxCADNER beta. Similarly, transcriptional activity of ER in CFs infected with AxCAER alpha or AxCAER beta was markedly enhanced and co-infection with AxCADNER beta abolished the effects. These results suggest that E2 inhibits CF growth and that both ER subtypes mediate the effect comparably and redundantly.
Biochemical and Biophysical Research Communications 12/2003; 311(2):454-9. · 2.48 Impact Factor
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Hitomi Nagayama,
Katsuaki Sato,
Mariko Morishita,
Kaoru Uchimaru,
Naoki Oyaizu,
Takeshi Inazawa,
Tomoko Yamasaki,
Makoto Enomoto, Takashi Nakaoka,
Tetsuya Nakamura,
Taira Maekawa,
Akifumi Yamamoto,
Shinji Shimada,
Toshiaki Saida,
Yutaka Kawakami,
Shigetaka Asano,
Kenzaburo Tani,
Tsuneo A Takahashi,
Naohide Yamashita
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ABSTRACT: We conducted a pilot study to assess the feasibility and efficacy of immunotherapy for stage IV malignant melanoma patients resistant to conventional therapies involving vaccination with mature dendritic cells (mDCs) combined with administration of low dose interleukin-2. Autologous monocytes were harvested from a single apheresis and cultured for 7 days with granulocyte-macrophage colony-stimulating factor and interleukin-4, yielding immature dendritic cells (iDCs), which were then cryopreserved until use. For 4 days prior to vaccination, iDCs were exposed to autologous tumour lysate combined with tumour necrosis factor-alpha to induce terminal differentiation into mDCs. Patients were then vaccinated weekly with 107 mDCs for 10 weeks and given 350-700 kIU of interleukin-2 three times per week. Of the 10 patients in the study, one showed stable disease, seven showed progressive disease, and two showed mixed responses, including partial tumour regression, and were therefore given 20 additional injections. Only minimal adverse events were noted, including localized skin reactions and mild fever (NIH-CTC grade 0-1). Median survival from the first vaccination was 240 days (range 31-735 days). In vitro, melanoma patient-derived dendritic cells (DCs) showed reduced cell surface expression of CD1a antigen on iDCs and reduced CD86 and HLA-DR expression on mDCs. In addition, antigen uptake, chemotaxis and antigen presentation were all attenuated in DCs from the patients. In summary, although improvement of clinical efficacy will require further research, autologous tumour lysate-pulsed monocyte-derived mDCs could be safely harvested, cryopreserved and administrated to patients without obvious complications.
Melanoma Research 11/2003; 13(5):521-30. · 2.19 Impact Factor
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ABSTRACT: We experienced a case of trichinellosis that had traveled in Kenya. The initial symptoms of the patient were myalgia and fever which started 2 weeks after she came back from Kenya, and blood examination showed marked eosinophilia (14,300/mm3) and elevated creatinine kinase (826IU/L). When we made serological diagnosis of trichinellosis 3 weeks after onset, symptoms started to resolve spontaneously and we observed the patient without any medication. As of 2 months after onset, symptoms and abnormal findings in laboratory data almost disappeared. The patient was speculated to be infected with trichinella spp. by incompletely cooked wild animal meats including alligator, zebra, pig, and ostrich during travel in Kenya. Since high incidence of trichinella infection in both wild and domestic animals has been reported in some areas of developing countries, travelers must be aware that raw or incompletely cooked animal meats can be source of trichinella spp.
Kansenshogaku zasshi. The Journal of the Japanese Association for Infectious Diseases 11/2003; 77(10):839-43.
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Tokumitsu Watanabe,
Masahiro Akishita, Takashi Nakaoka,
Koichi Kozaki,
Yukiko Miyahara,
Hong He,
Yumiko Ohike,
Teruhiko Ogita,
Satoshi Inoue,
Masami Muramatsu,
Naohide Yamashita,
Yasuyoshi Ouchi
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ABSTRACT: It has been demonstrated that 17beta-estradiol (E2) has an inhibitory effect on the proliferation of vascular smooth muscle cells (VSMCs) through an estrogen receptor (ER)-dependent pathway. Both ER subtypes, classical ER (ERalpha) and the newly identified ER subtype (ERbeta), are expressed in VSMCs. However, it remains unknown which receptor plays the critical role in the inhibitory effect on VSMC proliferation.
We constructed replication-deficient adenoviruses bearing the coding region of human ERalpha, ERbeta, and the dominant-negative form of ERbeta (designated AxCAERalpha, AxCAERbeta, and AxCADNERbeta, respectively). Prior to infection with the adenoviruses, 100 nmol/l E2 attenuated DNA synthesis by up to 14% and transactivated the estrogen-induced expression of the desired mRNA in rat VSMCs. This was accompanied by increased transcriptional activity of estrogen responsive element in response to E2, and the increase was comparable between AxCAERalpha and AxCAERbeta. When VSMCs were infected with AxCAERbeta at a multiplicity of infection of 5 or higher, DNA synthesis as well as cell number decreased by 50% in response to E2, and the effect was abolished by co-infection with AxCADNERbeta. In contrast, when VSMCs were infected with AxCAERalpha, the reduction in DNA synthesis was minimal.
Our results indicate that ERbeta is more potent than ERalpha in the inhibitory effect on VSMC proliferation.
Cardiovascular Research 10/2003; 59(3):734-44. · 6.06 Impact Factor
