Biomedical subjects

Manabu Mochizuki

Publications and source records attributed to Manabu Mochizuki.

At least 19 recordsLinked to original sources

PubMed2006

Tailor-made tricalcium phosphate bone implant directly fabricated by a three-dimensional ink-jet printer.

Rapid prototyping (RP) is a molding technique that builds a three-dimensional (3D) model from computer-aided design (CAD) data. We fabricated new tailor-made bone implants (TIs) from alpha-tricalcium phosphate powder using an RP ink-jet printer based on computed tomography (CT) data, and evaluated their safety and efficacy. CT data of the skulls of seven beagle dogs were obtained and converted to CAD data, and bone defects were virtually made in the skull bilaterally. TIs were designed to fit the defects and were fabricated using the 3D ink-jet printer with six horizontal cylindrical holes running through the implants, designed for possible facilitation of vascular invasion and bone regeneration. As a control, hydroxyapatite implants (HIs) were cut manually from porous hydroxyapatite blocks. Then, craniectomy was performed to create real skull defects, and TIs and HIs were implanted. After implantation, CT was performed regularly, and the animals were euthanized at 24 weeks. No major side effects were observed. CT analysis showed narrowing of the cylindrical holes; bony bridging between the implants and the temporal bone was observed only for TIs. Histological analysis revealed substantial new bone formation inside the cylindrical holes in the TIs, while mainly connective tissues invaded the porous structures in HIs. Bone marrow was observed only in TIs. Osteoclasts were seen to resorb regenerated bone from inside the cylindrical holes and to invade and probably resorb the TIs. These data suggest that TIs are a safe and effective bone substitute, possessing osteoconductivity comparable with that of HIs.

Animals↗

PubMed2006

Retinal and ciliary body pigment epithelium suppress activation of T lymphocytes via transforming growth factor beta.

The ocular microenvironment is immunosuppressive and anti-inflammatory. Pigment epithelial (PE) cells isolated from the eye possess a new property of suppressing T cell receptor-dependent activation of T cells in vitro. This property depends on their capacity to produce cell-surface and soluble inhibitory molecules. The iris pigment epithelia (IPE) do so through direct cell-to-cell contact with naïve T cells, and this suppressive contact is mediated by interactions between B7 and membrane-bound TGFbeta that are expressed constitutively on IPE. We have now examined whether other ocular PE cells, e.g., retinal pigment epithelia (RPE) and ciliary body pigment epithelia (CBPE), have a similar suppressive property by a similar process. We have found that RPE and CBPE significantly suppress the activation of bystander T cells via soluble inhibitory factors. RPE and CBPE secrete different soluble inhibitory factors including TGFbeta1 and TGFbeta2. Although IPE cells suppress the activation of bystander T cells by membrane-bound TGFbeta, the RPE and CBPE do so by soluble forms of active TGFbeta through mechanisms independent of cell contact. These ocular PE cells are capable modifying T cell function by enhancing production of regulatory cytokines including TGFbeta. We propose that this mechanism of suppression via TGFbeta ensures that soluble active TGFbeta is released into the ocular microenvironment in order to create the immune privilege of the posterior segment of the eye.

Animals↗

PubMed2006

In vitro and in vivo characterization of iris pigment epithelial cells cultured on amniotic membranes.

PURPOSE: To determine whether human amniotic membranes (AMs) can induce human and rat iris pigment epithelial (IPE) cells grown on them to develop characteristics of RPE cells in situ better than IPE cells grown on plastic plates, and to determine whether subretinal transplantation of IPE cell sheets grown on AMs can protect photoreceptor cells in dystrophic Royal College of Surgeons (RCS) rats. METHODS: IPE cells from humans and Long-Evans rats were cultured on the basement membrane side of dispase-treated AMs. Two weeks after seeding, ultrastructural changes were evaluated by transmission electron microscopy, and the level of expression of several genes present in differentiated retinal pigment epithelial (RPE) cells was determined by real time PCR and western blotting. IPE cell sheets cultured on AMs were transplanted into the subretinal space of 4-week-old RCS rats, and eyes were analyzed histologically 12 weeks after grafting. RESULTS: IPE cells cultured on AMs showed ultrastructural features like intercellular junctions, similar to RPE cells in situ. IPE cells grown on AMs had a greater upregulation in the expression of genes important for the function of differentiated RPE cells (e.g., pigment epithelium-derived factor [PEDF], RPE65, bestrophin, VEGF, and BDNF) than IPE cells grown on plastic plates. The number of photoreceptors present in RCS rats after subretinal transplantation of IPE cell sheets grown on AMs was significantly higher than that of sham injected rats and rats receiving transplantation of AMs without IPE cells. CONCLUSIONS: The more advanced degree of differentiation of IPE cells grown on AMs indicates that AMs are a better substrate to culture IPE cells than plastic plates. This was supported by the greater protection of photoreceptors of RCS rats when IPE cell sheets cultured on AMs were transplanted in the subretinal space.

Amnion↗

PubMed2006

Estimation of visual outcome without treatment in patients with subfoveal choroidal neovascularization in pathologic myopia.

OBJECTIVE: To clarify prognostic factors of long-term visual outcome without treatment in patients with myopic choroidal neovascularization (CNV) and estimate a regression equation to predict visual acuity at 5 years after CNV onset. METHODS: Fifty-four eyes of 54 consecutive patients with high myopia and subfoveal CNV who did not receive treatment were identified using clinical records from 1988 to 2004. Photodynamic therapy not approved for myopic CNV in Japan during this period. The association of best-corrected visual acuity (BCVA) at 5 years after CNV onset with patient age, refractive error, axial length, initial BCVA, myopic retinopathy grade, duration of persistent hemorrhage, CNV size at onset, and size of hemorrhage around the CNV was analyzed using Spearman's correlation and multiple linear regression analysis. RESULTS: BCVA at 5 years after onset was significantly associated with patient age, CNV size, and initial BCVA (P<0.05, Spearman's correlation). The regression equation estimating BCVA at 5 years after CNV onset was based on age and initial BCVA (R2=0.43). When subjects were divided into groups according to age (<40 and >or=40 years), CNV size, axial length and duration of persistent hemorrhage influenced BCVA at 5 years in patients under 40 years; while only initial BCVA was an influence in those at least 40 years old. CONCLUSIONS: We developed a linear predictive model to estimate BCVA at 5 years after onset of myopic CNV without treatment based on the identified prognostic factors. This information might be important for managing patients with myopic CNV.

Adult↗

PubMed2006

Choroidal vein and artery occlusion following photodynamic therapy in eyes with pathologic myopia.

PURPOSE: To describe highly myopic patients in which either the large choroidal veins or arteries were occluded following PDT treatment. METHODS: Demographic features of two highly myopic patients in which large choroidal vessels were occluded at 1 week following PDT, among a total of 23 patients who received PDT due to choroidal neovascularization (CNV) caused by high myopia, were demonstrated. RESULTS: In case 1, ICG angiography demonstrated the complete occlusion of several large choroidal veins within the laser-applied area at 1 week after PDT. In case 2, ICG angiography demonstrated an occlusion of choroidal artery emanating from beneath the CNV. One month after PDT, re-perfusion of the occluded choroidal vessels was noted in both patients. CONCLUSION: Occlusion of choroidal veins has never been reported in eyes with CNV treated by verteporfin therapy. Although further studies are necessary, occlusion of large choroidal vessels might happen in highly myopic eyes more commonly than expected. ICG angiography at 1 week after PDT was useful to evaluate the impact of PDT on large choroidal vessels.

Aged↗

PubMed2006

SUV39H1 interacts with HTLV-1 Tax and abrogates Tax transactivation of HTLV-1 LTR.

BACKGROUND: Tax is the oncoprotein of HTLV-1 which deregulates signal transduction pathways, transcription of genes and cell cycle regulation of host cells. Transacting function of Tax is mainly mediated by its protein-protein interactions with host cellular factors. As to Tax-mediated regulation of gene expression of HTLV-1 and cellular genes, Tax was shown to regulate histone acetylation through its physical interaction with histone acetylases and deacetylases. However, functional interaction of Tax with histone methyltransferases (HMTase) has not been studied. Here we examined the ability of Tax to interact with a histone methyltransferase SUV39H1 that methylates histone H3 lysine 9 (H3K9) and represses transcription of genes, and studied the functional effects of the interaction on HTLV-1 gene expression. RESULTS: Tax was shown to interact with SUV39H1 in vitro, and the interaction is largely dependent on the C-terminal half of SUV39H1 containing the SET domain. Tax does not affect the methyltransferase activity of SUV39H1 but tethers SUV39H1 to a Tax containing complex in the nuclei. In reporter gene assays, co-expression of SUV39H1 represses Tax transactivation of HTLV-1 LTR promoter activity, which was dependent on the methyltransferase activity of SUV39H1. Furthermore, SUV39H1 expression is induced along with Tax in JPX9 cells. Chromatin immunoprecipitation (ChIP) analysis shows localization of SUV39H1 on the LTR after Tax induction, but not in the absence of Tax induction, in JPX9 transformants retaining HTLV-1-Luc plasmid. Immunoblotting shows higher levels of SUV39H1 expression in HTLV-1 transformed and latently infected cell lines. CONCLUSION: Our study revealed for the first time the interaction between Tax and SUV39H1 and apparent tethering of SUV39H1 by Tax to the HTLV-1 LTR. It is speculated that Tax-mediated tethering of SUV39H1 to the LTR and induction of the repressive histone modification on the chromatin through H3 K9 methylation may be the basis for the dose-dependent repression of Tax transactivation of LTR by SUV39H1. Tax-induced SUV39H1 expression, Tax-SUV39H1 interaction and tethering to the LTR may provide a support for an idea that the above sequence of events may form a negative feedback loop that self-limits HTLV-1 viral gene expression in infected cells.

Base Sequence↗

PubMed2006

Characteristics of peripapillary detachment in pathologic myopia.

OBJECTIVE: To evaluate the prevalence and clinical features of a newly recognized peripapillary lesion specific to high myopia, peripapillary detachment in pathologic myopia (PDPM), in a large series of patients with high myopia. METHODS: Three hundred twenty-four patients (632 eyes) with high myopia were enrolled in this study. We examined the prevalence, range, fluorescein and indocyanine green angiographic findings, and optical coherence tomography findings of PDPM for these patients. Visual field testing (Goldmann kinetic perimetry and the Humphrey 30-2 program) was also performed in the patients with PDPM. RESULTS: Peripapillary detachment in pathologic myopia was identified in 31 of 632 highly myopic eyes (4.9%). The optical coherence tomographic scan across the PDPM lesion revealed a localized detachment of retinal pigment epithelium adjacent to the optic nerve. Although PDPM was always situated adjacent to the inferior edge of the optic disc, in some patients it surrounded almost the entire optic disc. There was a steep excavation of the inferior myopic conus adjacent to the PDPM, and the inferotemporal retinal vein was markedly bent at the transition from the PDPM to the excavated myopic conus. Glaucomatous visual field defects were frequently detected in eyes with PDPM (71.0%). CONCLUSIONS: The findings of this study indicate that PDPM is not uncommon among highly myopic eyes. Although its pathogenesis and pathologic significance require further classification, PDPM might be another indicator of visual field defects in high myopia.

Adolescent↗

PubMed2006

Natural course of macular retinoschisis in highly myopic eyes without macular hole or retinal detachment.

PURPOSE: To investigate the natural course of macular retinoschisis in highly myopic patients. DESIGN: A prospective, observational case series. METHODS: Seven consecutive patients (eight eyes) with macular retinoschisis without macular hole or retinal detachment determined by optical coherence tomography (OCT) were followed at least two years without surgical intervention. The changes in the OCT findings and best-corrected visual acuity (BCVA) were analyzed. RESULTS: During follow-up, two eyes that had vitreoretinal adhesions developed a macular hole, one with and one without retinal detachment (RD), and two eyes without detectable vitreoretinal adhesion developed RD without a macular hole. BCVA in these four eyes was significantly reduced. The remaining four eyes did not develop complications, although the thickness of the macula increased significantly. CONCLUSIONS: These findings suggest that macular retinoschisis might be a progressive condition, and that complications appeared to be related to the presence of vitreoretinal tractions.

Aged↗

PubMed2006

Pattern-reversal visual evoked potentials in patients with human T-lymphotropic virus type 1 uveitis.

PURPOSE: The purpose of this study was to evaluate the possible injury in the optic pathway by measuring P100 peak latency of pattern-reversal visual evoked potentials (PVEPs) in patients with human T-lymphotropic virus type 1 uveitis (HU). METHODS: The P100 peak latency of PVEP was measured during the period without macular abnormalities observed by fluorescein angiography in 23 patients (46 eyes) with HU and 24 patients (48 eyes) with Vogt-Koyanagi-Harada disease (VKH) with a corrected visual acuity of 20/25 or more. To determine the normal upper limit of P100 peak latency, PVEPs were measured in 31 normal subjects (31 eyes). In addition, in the HU patients, the serum anti-HTLV-1 antibody titer was measured by particle agglutination assay within 3 months of PVEP recording, and the period of HU was retrospectively surveyed. RESULTS: Delayed latency was observed in 4 (7 eyes) of the 23 patients (46 eyes) with HU but none of the 24 patients (48 eyes) with VKH. All four patients with delayed latency showed a serum anti-HTLV-1 antibody titer of more than x4000. The HU period in the HU patients was 0.2-14.0 years, and the HU periods in the four patients with delayed latency were 0.8, 2.7, 4.2, and 14.0 years, respectively. CONCLUSIONS: We measured pattern-reversal visual evoked potentials and observed delayed P100 peak latency in 7 of the 46 eyes in 4 (17.4%) of the 23 HU patients. This suggests injury in the optic pathway including the optic nerve by HTLV-1 in some patients with HU. In the future, consideration should also be given to the possible development of optic neuropathy due to HTLV-1.

Adolescent↗

PubMed2006

The effects of propofol, isoflurane and sevoflurane on vecuronium infusion rates for surgical muscle relaxation in dogs.

OBJECTIVE: To compare the constant rate infusion (CRI) of vecuronium required to maintain a level of neuromuscular blockade adequate for major surgeries, e.g. thoracotomy or laparotomy, in dogs anaesthetized with a CRI of fentanyl and either propofol, isoflurane or sevoflurane. STUDY DESIGN: Prospective, randomized, cross-over study. ANIMALS: Thirteen male beagles (age, 9-22 months; body mass 6.3-11.3 kg). MATERIALS AND METHODS: Dogs were anaesthetized with propofol (24 mg kg(-1) hour(-1) IV CRI; group P), isoflurane (1.3% end-tidal concentration; group I) or sevoflurane (2.3% end-tidal concentration; group S) with fentanyl (5 microg kg(-1) hour(-1) IV, CRI). Sixty to seventy minutes after induction of anaesthesia, vecuronium was administered at a rate of 0.4, 0.3 and 0.2 mg kg(-1) hour(-1) in groups P, I and S respectively. To determine the degree of neuromuscular block, a peripheral nerve was stimulated electrically using the train-of-four (TO4) stimulus pattern. Evoked muscle contractions were evaluated using a neuromuscular monitoring device. Once the TO4 ratio reached 0, the continuous infusion rate was decreased and adjusted to maintain a TO4 count of 1. Continuous infusion was continued for 2 hours. The infusion rate of vecuronium was recorded 20, 40, 60, 80, 100 and 120 minutes after the start of infusion. RESULTS: The mean continuous infusion rates of vecuronium during stable infusion were 0.22 +/- 0.04 (mean +/- SD), 0.10 +/- 0.02 and 0.09 +/- 0.02 mg kg(-1) hour(-1) in groups P, I and S respectively. There were statistically significant differences between the rates in groups P and I and between the rates in groups P and S. Conclusions and clinical relevance In healthy dogs, the recommended maintenance infusion rate of vecuronium is 0.2 mg kg(-1) hour(-1) under CRI propofol-fentanyl anaesthesia and 0.1 mg kg(-1) hour(-1) during CRI fentanyl-isoflurane or sevoflurane anaesthesia.

Anesthesia, General↗

PubMed2006

Pharmacokinetics of fentanyl after single intravenous injection and constant rate infusion in dogs.

OBJECTIVE: To determine the plasma concentration and define the pharmacokinetic characteristics of fentanyl (10 microg kg(-1)) administered as a single intravenous (IV) injection followed by: (a) no further drug; or (b) a constant rate infusion (CRI) of fentanyl 10 microg kg(-1) hour(-1) lasting 1, 3 or 4 hours in dogs. Animals Fourteen healthy adult beagles (seven males and seven females). EXPERIMENTAL DESIGN: Randomized cross-over design. MATERIALS AND METHODS: Dogs were randomly assigned to four treatment groups. Drugs were administered to each dog in a randomized cross-over design with at least a 14-day washout interval between experiments. All dogs received an IV loading dose of fentanyl (10 microg kg(-1)). One group received no further fentanyl. In others, the loading dose was followed by a CRI of fentanyl (10 microg kg(-1) hour(-1)) for 1, 3 or 4 hours. Blood samples were collected and plasma fentanyl concentrations determined using high-performance liquid chromatography-mass spectrometry. Plasma pharmacokinetic estimates were obtained by plotting plasma concentrations versus time data and by fitting the change in concentration to a pharmacokinetic model, using a purpose-built program written by the Graduate School of Pharmaceutical Sciences (Kyoto University) in Visual Basic (VBA) on Excel (Microsoft Corporation). RESULTS: Plasma fentanyl concentration decreased rapidly after single IV injection: the plasma concentration-time curve best fitted a two-compartment model. Pharmacokinetic variables for IV injection were characterized by a short distribution half-time (t1/2alpha was 4.5 minutes), a relatively long elimination half time (t1/2beta was 45.7 minutes), a large volume of distribution (approximately 5 L kg(-1)) and high total body clearance (77.9 mL minute(-1) kg(-1)). Stable plasma fentanyl levels were obtained in all CRI groups although pharmacokinetic variables were influenced by the duration of administration. CONCLUSIONS AND CLINICAL RELEVANCE: While this study clarified the pharmacokinetic features of rapid IV fentanyl injection and CRI in dogs, the plasma concentration achieving analgesia was not and so further research is needed. Further studies on the effects of other sedatives and/or anaesthetics on fentanyl's disposition are also required as the drug is commonly used with other agents.

Anesthesia, General↗

PubMed2006

Trans-Tenon's retrobulbar triamcinolone infusion for myopic choroidal neovascularization.

PURPOSE: To report the effects of trans-Tenon's retrobulbar injection of triamcinolone acetonide for subfoveal and juxtafoveal choroidal neovascularization (CNV) caused by pathological myopia. METHODS: Eleven consecutive patients (11 eyes) with myopic CNV were treated with trans-Tenon's retrobulbar injection of triamcinolone acetonide. Each patient received a single injection. Evaluation included best corrected visual acuity (BCVA) measurements, fluorescein fundus angiography, retinal oedema examined by optical coherence tomography (OCT), and retinal sensitivity using scanning laser ophthalmoscopy (SLO) at the initial examination and at 6 and 12 months after treatment. RESULTS: At 6 months after treatment, BCVA had improved by at least two ETDRS lines in eight eyes, and remained unchanged in three eyes. No eye showed worsening of VA by two or more ETDRS lines. At 12 months, BCVA had improved by at least two ETDRS lines in 10 eyes and remained unchanged in only one eye. The size of the CNV decreased in all patients after treatment. Fluorescein fundus angiography revealed an absence of dye leakage in the late angiographic phase. Optical coherence tomography revealed decreased retinal oedema in all patients and SLO microperimetry revealed an increase in retinal sensitivity in seven eyes, at both 6 and 12 months after treatment. Chorioretinal atrophy developed around the CNV in 10 eyes at 6 months and in all eyes at 12 months after treatment. CONCLUSIONS: Trans-Tenon's retrobulbar injection of triamcinolone acetonide for CNV resulting from pathological myopia appears to be relatively safe and to have a good visual outcome, although a longterm follow-up study in a larger series of patients is necessary.

Adult↗

PubMed2006

Cytokine profile in aqueous humor and sera of patients with infectious or noninfectious uveitis.

PURPOSE: To determine the cytokine expression profile at the protein level in aqueous humor (AqH) and sera of patients with uveitis. METHODS: Patients with various clinical entities of strictly diagnosed infectious or noninfectious uveitis were tested. AqH and sera were collected from patients with uveitis. AqH was also collected during surgery from patients with cataract, as control specimens. Interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, and interleukin (IL)-2, -4, -5, and -10 were measured from nondiluted samples simultaneously, with microparticle-based flow cytometric analysis. RESULTS: In AqH IFN-gamma was the most abundant cytokine in both infectious (mean, 3240.5 pg/mL) and noninfectious (mean, 115.6 pg/mL) uveitis, and IL-10 was the second (mean, 402.1 pg/mL, infectious uveitis; 7.5 pg/mL, noninfectious uveitis). The expression level of other cytokines in AqH was generally higher in infectious uveitis than in noninfectious uveitis, but the levels were lower than that of IL-10. There was no remarkable difference, however, in the cytokine expression pattern in AqH of the different clinical entities of uveitis. Sera from patients with noninfectious uveitis contained IFN-gamma (mean, 45.0 pg/mL), but the other serum cytokines in both types of uveitis were low or under the detectable level. CONCLUSIONS: IFN-gamma is the most abundant cytokine in infectious and noninfectious uveitis, with a remarkable difference between the two groups. The data suggest that cytokines in AqH of infectious uveitis are locally produced, whereas in noninfectious uveitis, IFN-gamma is produced both in the eye and the peripheral blood.

Adult↗

PubMed2006

Detection of the bcl-2 t(14;18) translocation and proto-oncogene expression in primary intraocular lymphoma.

PURPOSE: Primary intraocular lymphoma (PIOL) is a diffuse large B cell lymphoma that initially infiltrates the retina, vitreous, or optic nerve head, with or without central nervous system involvement. This study examined the expression of the bcl-2 t(14;18) translocation, the bcl-10 gene, and high expression of bcl-6 mRNA in PIOL cells. METHODS: Microdissection and PCR analysis were used to examine vitreous specimens in patients with PIOL for the presence of bcl-2 t(14;18) translocations, the bcl-10 gene, and expression of bcl-6 mRNA. A medical record review was also conducted to determine whether the bcl-2 t(14;18) translocation correlated with prognosis. RESULTS: Forty of 72 (55%) PIOL patients expressed the bcl-2 t(14;18) translocation at the major breakpoint region. Fifteen of 68 (22%) patients expressed the translocation at the minor cluster region. The bcl-10 gene was detected in 6 of 26 (23%) patients, whereas 4 of 4 (100%) PIOL patients expressed higher levels of bcl-6 mRNA compared with inflammatory lymphocytes. An analysis of clinical outcome in 23 PIOL patients revealed no significant association between bcl-2 t(14;18) translocations and survival or relapse. However, patients with the translocation were significantly younger. CONCLUSIONS: PIOL has unique molecular patterns of bcl-2, bcl-10, and bcl-6 when compared with other systemic lymphomas. This study lays the foundation for future studies aimed at exploring the genotypic classification of PIOL based on the quantitative molecular framework of gene expression profiling, with the goal of providing useful adjuncts to the pathologic diagnosis of this complex disease.

Adaptor Proteins, Signal Transducing↗

PubMed2006

B7+ iris pigment epithelial cells convert T cells into CTLA-4+, B7-expressing CD8+ regulatory T cells.

PURPOSE: To determine whether iris PE (IPE) promotes the generation of regulatory T-cells (Tregs) with cell contact via B7-2/CTLA-4 interactions. METHODS: T cells were cocultured with IPE cells obtained from eyes of normal and B7-deficient mice, x-irradiated, and used as regulators. IPE T regulator cells (IPE Tregs) of normal and CD28- or CTLA-4-deficient mice were established. Target bystander T cells were established from normal splenic T cells with anti-CD3 antibodies. T-cell activation was assessed for proliferation by [3H]-thymidine incorporation. Neutralizing anti-B7-1 and/or B7-2 antibodies, anti-CTLA-4 antibodies, CTLA-4-Ig fusion proteins were used to abolish regulatory function. IPE-exposed CD8+ T cells were evaluated for expression of B7, CTLA-4, and Foxp3 by using RT-PCR and flow cytometry. CD8+ IPE Tregs were depleted of B7-2+ and CTLA-4+ T cells and assayed for suppressive activity by adding them to bystander T cells. RESULTS: T cells acquired T regulatory activity when exposed to cultured IPE. Ciliary body PE cells did not promote conversion of T cells into Tregs. IPE converted CD8+, but not CD4+, T cells into Tregs by direct cell contact. In the conversion, IPE and responding T cells must both express endogenously synthesized B7-1 and B7-2, and the T cells must also express CTLA-4. Expression of CD28 molecules was not necessary for Treg generation. In addition, the CD8+ Tregs that fully suppress activation of bystander T cells expressed Foxp3. CONCLUSIONS: IPE cells promote conversion of T cells into Tregs solely through a contact-dependent mechanism. T cells exposed to IPE cells acquire full regulatory capacity.

Animals↗

PubMed2006

Ocular infiltrating CD4+ T cells from patients with Vogt-Koyanagi-Harada disease recognize human melanocyte antigens.

PURPOSE: To determine whether patients with Vogt-Koyanagi-Harada (VKH) disease have immune responses specific to the melanocyte antigens tyrosinase and gp100. METHODS: T-cell clones (TCCs) were established from cells infiltrating the aqueous humor and from peripheral blood mononuclear cells (PBMCs) of patients with VKH. The target cells were LDR4-transfected cells (HLA-DRB1*0405). The TCCs were cocultured with LDR4 in the presence of tyrosinase (tyrosinase450-462: SYLQDSDPDSFQD), gp100 (gp100(44-59): WNRQLYPEWTEAQRLD), or a control peptide. The immune response was evaluated by cytokine production. The responding melanocyte peptide-specific VKH-TCCs were characterized by an immunofluorescence method with flow cytometry. A search was made for molecular mimicry among tyrosinase450-462, gp100(44-59), and exogenous antigens, such as viruses, by database screening. RESULTS: Cells infiltrating the eye and PBMCs in HLA-DR4+ (HLA-DRB1*0405, 0410) patients with VKH contained a population of CD4+ T lymphocytes that recognized tyrosinase and gp100 peptides and produced RANTES and IFN-gamma in response to the two peptides. The T cells were active memory Th1-type lymphocytes, and they recognized the tyrosinase peptide and produced IFN-gamma in response to HLA-DRB1*0405+ melanoma cells. Cytomegalovirus envelope glycoprotein H (CMV-egH290-302) had high amino acid homology with the tyrosinase peptide. In addition, some of the VKH-TCCs recognized CMV-egH290-302 peptide, as well as the tyrosinase peptides. CONCLUSIONS: In VKH there are tyrosinase and gp100 peptide-specific T cells that can mediate an inflammatory response. Such melanocyte antigen-specific T cells could be associated with the cause and pathology of VKH disease.

CD4-Positive T-Lymphocytes↗

PubMed2006

Retinoids induce growth inhibition and apoptosis in mast cell tumor cell lines.

Retinoids are well recognized as promising antitumor agents in humans. However, there have only been a few reports about the effect of retinoids in canine cancers. To investigate the antitumor effect of retinoids on mast cell tumors (MCT), inhibitory effect on cell growth and induction of apoptosis were examined in vitro. Although sensitivity of these cells differed among the cells, the growth of three MCT cell lines (CoMS, CM-MC and VI-MC) were inhibited dose dependently when they were treated with retinoids. FACS analysis of PI-stained nuclei revealed an apoptotic fraction in CM-MC cells about 30% when treated with retinoids, while those of control cells were less than 5%. Caspase-3 activation was observed after retinoid treatment in CM-MC cells. This was confirmed by inhibiting the retinoid-induced apoptosis using the pan-caspase inhibitor, ZVAD-FMK. Both retinoid receptors, RARs and RXRs, were detected by immunoprecipitation followed by western blot analysis in all the three MCT cells. These data suggests that retinoids inhibit the growth of MCTs partly through apoptosis, and this growth inhibition by retinoids may be mediated by RARs and RXRs. We conclude that retinoid may be a potential adjunctive chemotherapeutic agent for the treatment of canine MCT.

Animals↗

PubMed2006

Suppressive effect of hyaluronan on chondrocyte apoptosis in experimentally induced acute osteoarthritis in dogs.

Dogs receiving anterior cruciate ligament transection (ACLT) were treated with either intravenous (IV) or intraarticular (IA) administration of hyaluronan (HA), and differences in appearance of chondrocyte apoptosis of the stifle joint were investigated. Chondrocyte apoptosis was detected using flow cytometry as well as by staining with TdT-mediated dUTP nick end labeling (TUNEL). The percentage of apoptotic chondrocytes in dogs with ACLT was significantly higher than that in intact (non-ACLT) dogs. Dogs treated with IA or IV injection of HA after ACLT had fewer apoptotic chondrocytes than non-treated dogs after ACLT. It was suggested that ACLT-induced apoptosis of chondrocytes was suppressed by HA administration of either IA or IV.

Animals↗