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Hisayuki Ueno - One of the best experts on this subject based on the ideXlab platform.
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suppression of macrophage infiltration into the Conjunctiva by clodronate liposomes in experimental immune mediated blepharoconjunctivitis
Cell Biology International, 2005Co-Authors: Atsuki Fukushima, Akemi Ozaki, Waka Ishida, Kazuyo Fukata, Nico Van Rooijen, Hisayuki UenoAbstract:Macrophages infiltrate the Conjunctiva in severe cases of allergic conjunctivitis (AC) such as atopic keratoconjunctivitis (AKC). We established experimental immune-mediated blepharoconjunctivitis (EC) in Brown Norway (BN) rats as a model for severe types of AC. We investigated whether macrophage infiltration in the Conjunctiva in this EC model is inhibited by clodronate liposomes (CL2MDP-lip). The numbers of ED1-positive but not ED2-positive macrophages in the Conjunctivas were increased by the induction of EC. SubConjunctival injection of CL2MDP-lip decreased the number of ED2-positive but not ED1-positive macrophages in the Conjunctivas of naive rats. CL2MDP-lip did not affect macrophages in the spleen. SubConjunctival injection of CL2MDP-lip into EC-developing BN rats decreased the number of ED2-positive macrophages at all the time points. ED1-positive cell infiltration was inhibited when treatment was administered just prior to OVA challenge. Intravenous injection of CL2MDP-lip decreased the number of ED2-positive cells in the Conjunctiva. Thus, we conclude that CL2MDP-lip inhibits infiltration of macrophages into the Conjunctiva within 24 h of antigen challenge.
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ag specific recognition activation and effector function of t cells in the Conjunctiva with experimental immune mediated blepharoconjunctivitis
Investigative Ophthalmology & Visual Science, 2003Co-Authors: Atsuki Fukushima, Akemi Ozaki, Waka Ishida, Kazuyo Fukata, Hisayuki UenoAbstract:PURPOSE. To investigate antigen (Ag) specificity, activation, and effector function of the Ag-specific T cells involved in the development of experimental immune-mediated blepharoconjunctivitis (EC), an experimental conjunctivitis. METHODS. EC was induced in Brown Norway rats by injection of ovalbumin (OVA)-specific T cells followed by OVA challenge with eye drops. Eyes, including the Conjunctivas, were harvested at different time points after challenge. The dependence of EC onset on the challenging Ag was assessed by challenge with an irrelevant Ag or stimulatory OVA peptides. To show the infiltration of transferred T cells into the Conjunctiva, T cells were labeled with 5-(and-6)-carboxyfluorescein diacetate succinimidyl ester (CFSE) before transfer. The activation of T cells in the Conjunctiva was assessed by measuring phosphorylation of Lck-associated molecules by Western blot analysis. Conjunctivas were also examined by immunohistochemistry and used for reverse transcription-polymerase chain reaction to determine the phenotype of the infiltrating cells and cytokine, chemokine, and chemokine receptor expression. To investigate infiltration of non Ag-specific T cells into the Conjunctiva, ragweed (RW)-primed lymphocytes were transferred into OVA-specific T-cell receptor transgenic (DOI1.10) mice. The mice were then challenged with RW and the Conjunctivas were harvested for immunohistochemistry to detect T cells derived from DO11.10 mice. RESULTS. EC was induced only when challenged with OVA protein or stimulatory OVA peptides, and CFSE-labeled transferred cells were found in the Conjunctiva. Phosphorylation of Lck and an 85-kDa Lck-associated molecule were observed in the Conjunctiva 6 hours after challenge. Many cytokines and chemokines began to be expressed at 6 hours, and individual expression patterns over time correlated well with the infiltration patterns of different inflammatory cells. In DO11.10 mice that received RW-primed lymphocytes, T cells derived from the recipient mice infiltrated the Conjunctiva after RW challenge. CONCLUSIONS. Ag-specific T cells initiate EC by first infiltrating the Conjunctiva, where they become activated by the specific Ag in the Conjunctiva.
Xinghuai Sun - One of the best experts on this subject based on the ideXlab platform.
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evaluation of age related changes in human palpebral Conjunctiva and meibomian glands by in vivo confocal microscopy
Cornea, 2011Co-Authors: Anji Wei, Jiaxu Hong, Xinghuai SunAbstract:PURPOSE To investigate the age-related changes in human palpebral Conjunctiva and meibomian glands by in vivo confocal microscopy. METHODS Forty-nine healthy volunteers (20 men and 29 women; mean age, 43.4 ± 22.7 years; range, 9-85 years) were recruited from the community. Laser scanning confocal microscopy was used to observe and measure the upper palpebral Conjunctiva and meibomian glands. Customized software was used to analyze the images. The quantitative parameters included the mean densities of goblet cells, Conjunctival basal epithelium, Langerhans cells, and meibomian glandular acinar units. RESULTS Mean densities of goblet cells, Conjunctival basal epithelium, Langerhans cells, and meibomian glandular acinar units were 1050 ± 495 cells per square millimeter, 2979 ± 510 cells per square millimeter, 32 ± 16 cells per square millimeter, and 71 ± 27 glands per square millimeter in total subjects, respectively. No statistically significant difference in these 4 parameters was observed between the male and female subjects. Significant negative correlations were noted between age and goblet cell density (r = -0.646; P < 0.0001) and meibomian glandular acinar unit density (r = -0.585; P < 0.0001). However, no significant correlation was detected between the densities of Conjunctival basal epithelium or Langerhans cell density and age. CONCLUSIONS Age-related changes under laser scanning confocal microscopy included decreased densities of goblet cells in human palpebral Conjunctiva and the acinar units in meibomian glands.
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age related changes of human Conjunctiva on in vivo confocal microscopy
British Journal of Ophthalmology, 2010Co-Authors: Wenqing Zhu, Jiaxu Hong, Tianyu Zheng, Xinghuai SunAbstract:Aims To analyse age-related changes of human Conjunctiva by in vivo laser scanning confocal microscopy (LSCM). Methods 80 healthy subjects were enrolled and divided into four groups according to age: A (0–19 years), B (20–39 years), C (40–59 years) and D (over 60 years). In vivo LSCM was performed on the subjects9 bulbar Conjunctivas, and the images were recorded. The morphology and densities of Conjunctival epithelial cells, goblet cells (GCs), dendritic cells (DCs) and the positive rate of Conjunctival microcysts were analysed. The diameters of subepithelial fibres in each group were measured as well. Results The morphology and the densities of both Conjunctival epithelial cells and GCs showed no significant age-related differences. The positive rates of Conjunctival microcysts were 30% (6/20), 50% (10/20), 60% (12/20) and 75% (15/20) in each group, showing a substantial increase with age (p=0.035). The densities of DCs decreased significantly with ageing (p=0.033). The diameters of subepithelial fibres reduced with age significantly also (p=0.000). Conclusions The age-related changes of Conjunctivas included decreasing densities of DCs as well as the degeneration of subepithelial structures. Even though no significant changes in GCs densities could be seen in the different age groups, the increasing rates of Conjunctival microcysts indicated that the cellular function of GCs declined while ageing.
Teruo Nishida - One of the best experts on this subject based on the ideXlab platform.
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role of structural cells of the cornea and Conjunctiva in the pathogenesis of vernal keratoconjunctivitis
Progress in Retinal and Eye Research, 2006Co-Authors: Naoki Kumagai, Ken Fukuda, Youichiro Fujitsu, Kazutaka Yamamoto, Teruo NishidaAbstract:Vernal keratoconjunctivitis (VKC) is a severe type of allergic Conjunctival disease characterized by the presence both of various corneal epithelial and stromal lesions as well as of Conjunctival proliferative changes such as giant papillae of the upper tarsal Conjunctiva and limbal lesions. These clinical findings as well as various pathophysiological characteristics of VKC are distinct from those of other types of ocular allergy and allergic diseases of other organs. The outer eye possesses specific allergological characteristics, one of which is communication between the cornea and Conjunctiva through a thin layer of tear fluid. Fibroblasts of the cornea and the Conjunctiva are activated by proinflammatory and T helper 2 (Th2) cell-derived cytokines. Corneal fibroblasts enhance ocular allergic reactions as a result of their activation-induced expression both of chemokines such as eotaxin and TARC as well as of adhesion molecules such as ICAM-1 and VCAM-1, all of which together promote the activation and infiltration of eosinophils and Th2 lymphocytes. In contrast, corneal epithelial cells suppress such reactions by physically separating corneal fibroblasts from bioactive substances in tear fluid. Exaggerated proliferation of and deposition of extracellular matrix by Conjunctival fibroblasts likely exacerbate Conjunctival inflammation. Restoration of an intact corneal epithelium and inhibition of the activities of corneal and Conjunctival fibroblasts may provide a basis for the development of new treatments for severe ocular allergic diseases such as VKC.
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differential distribution of subchains of the basement membrane components type iv collagen and laminin among the amniotic membrane cornea and Conjunctiva
Cornea, 1999Co-Authors: Ken Fukuda, Taiichiro Chikama, Masatsugu Nakamura, Teruo NishidaAbstract:Purpose Amniotic membrane transplantation has been reported to be an effective surgical procedure for the reconstruction of the anterior segment of the eye. To understand better the function of transplanted amniotic membrane, we compared the distributions of subchains of type IV collagen and laminin in the amniotic membrane to those in the cornea and Conjunctiva. Methods Five human corneas with Conjunctivas and three human amniotic membranes were frozen in OCT compound. Cryosections were cut with a cryostat and stained by an indirect immunofluorescent microscopy. We used antibodies of the collagen alpha2(IV) and alpha5(IV) subchains, laminin-1, laminin-5, fibronectin, and type VII collagen. Results In the Conjunctival basement membrane and the amniotic membrane, fluorescence was evident for collagen alpha2(IV) but not for collagen alpha5(IV). By contrast, in the corneal basement membrane, fluorescence was apparent for the collagen alpha5(IV) subchain but not for the collagen alpha2(IV) subchain. Laminin-1, laminin-5, fibronectin, and type VII collagen were present in all the basement membranes examined. Conclusion The distribution of alpha subchains of type IV collagen in the amniotic membrane was identical to that in the Conjunctiva but different from that in the cornea. No difference in the distribution pattern of other basement membrane components was observed. These results demonstrate that the basement membrane of the amniotic membrane and the Conjunctiva might share the same components; therefore, the amniotic membrane might be useful as a replacement for the basement membrane of the Conjunctiva.
Nicola Pritchard - One of the best experts on this subject based on the ideXlab platform.
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confocal microscopy of the bulbar Conjunctiva in contact lens wear
Cornea, 2010Co-Authors: Nathan Efron, Munira Aldossari, Nicola PritchardAbstract:Purpose: The aim of this study was to investigate the capabilities of laser scanning confocal microscopy (LSCM) for undertaking qualitative and quantitative investigations of the response of the bulbar Conjunctiva to contact lens wear. Methods: LSCM was used to observe and measure morphological characteristics of the bulbar Conjunctiva of 11 asymptomatic soft contact lens wearers and 11 healthy volunteer subjects (controls). Results: The appearance of the bulbar Conjunctiva is consistent with known histology of this tissue based on light and electron microscopy. The thickness of the bulbar Conjunctival epithelium of lens wearers (30.9 ± 1.1 μm) was less than that of controls (32.9 ± 1.1 μm) (P < 0.0001). Superficial and basal bulbar Conjunctival epithelial cell densities in contact lens wearers were 91% and 79% higher, respectively, than that in controls (P < 0.0001). No difference was observed in goblet and Langerhans cell density between lens wearers and controls. Conjunctival microcysts were observed in greater numbers, and were larger in size, in lens wearers compared with controls. Conclusions: The effects of contact lens wear on the human bulbar Conjunctiva can be investigated effectively at a cellular level using LSCM. The observations in this study suggest that contact lens wear can induce changes in the bulbar Conjunctiva such as epithelial thinning and accelerated formation and enlargement of microcysts, increased epithelial cell density, but has no impact on goblet or Langerhans cell density.
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In vivo confocal microscopy of the bulbar Conjunctiva
Clinical and Experimental Ophthalmology, 2009Co-Authors: Nathan Efron, Munira Al-dossari, Nicola PritchardAbstract:Background: The aim of this work is to develop a more complete qualitative and quantitative understanding of the in vivo histology of the human bulbar Conjunctiva. Methods: Laser scanning confocal microscopy (LSCM) was used to observe and measure morphological characteristics of the bulbar Conjunctiva of 11 healthy human volunteer subjects. Results: The superficial epithelial layer of the bulbar Conjunctiva is seen as a mass of small cell nuclei. Cell borders are sometimes visible. The light grey borders of basal epithelial cells are clearly visible, but nuclei can not be seen. The Conjunctival stroma is comprised of a dense meshwork of white fibres, through which traverse blood vessels containing cellular elements. Orifices at the epithelial surface may represent goblet cells that have opened and expelled their contents. Goblet cells are also observed in the deeper epithelial layers, as well as Conjunctival microcysts and mature forms of Langerhans cells. The bulbar Conjunctiva has a mean thickness of 32.9 ± 1.1 µm, and a superficial and basal epithelial cell density of 2212 ± 782 and 2368 ± 741 cells/mm2, respectively. Overall goblet and mature Langerhans cell densities are 111 ± 58 and 23 ± 25 cells/mm2, respectively. Conclusions: LSCM is a powerful technique for studying the human bulbar Conjunctiva in vivo and quantifying key aspects of cell morphology. The observations presented here may serve as a useful marker against which changes in Conjunctival morphology due to disease, surgery, drug therapy or contact lens wear can be assessed.
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In Vivo Confocal Microscopy of the Bubar Conjunctiva
Clinical and Experimental Ophthalmology, 2009Co-Authors: Nathan Efron, Munira Al-dossari, Nicola PritchardAbstract:Background: The aim of this work is to develop a more complete qualitative and quantitative understanding of the in vivo histology of the human bulbar Conjunctiva. Methods: Laser scanning confocal microscopy (LSCM) was used to observe and measure morphological characteristics of the bulbar Conjunctiva of 11 healthy human volunteer subjects. Results: The superficial epithelial layer of the bulbar Conjunctiva is seen as a mass of small cell nuclei. Cell borders are sometimes visible. The light grey borders of basal epithelial cells are clearly visible, but nuclei can not be seen. The Conjunctival stroma is comprised of a dense meshwork of white fibres, through which traverse blood vessels containing cellular elements. Orifices at the epithelial surface may represent goblet cells that have opened and expelled their contents. Goblet cells are also observed in the deeper epithelial layers, as well as Conjunctival microcysts and mature forms of Langerhans cells. The bulbar Conjunctiva has a mean thickness of 32.9 1.1 mm, and a superficial and basal epithelial cell density of 2212 782 and 2368 741 cells/ mm2, respectively. Overall goblet and mature Langerhans cell densities are 111 58 and 23 25 cells/mm2, respectively. Conclusions: LSCM is a powerful technique for studying the human bulbar Conjunctiva in vivo and quantifying key aspects of cell morphology. The observations presented here may serve as a useful marker against which changes in Conjunctival morphology due to disease, surgery, drug therapy or contact lens wear can be assessed.
Atsuki Fukushima - One of the best experts on this subject based on the ideXlab platform.
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suppression of macrophage infiltration into the Conjunctiva by clodronate liposomes in experimental immune mediated blepharoconjunctivitis
Cell Biology International, 2005Co-Authors: Atsuki Fukushima, Akemi Ozaki, Waka Ishida, Kazuyo Fukata, Nico Van Rooijen, Hisayuki UenoAbstract:Macrophages infiltrate the Conjunctiva in severe cases of allergic conjunctivitis (AC) such as atopic keratoconjunctivitis (AKC). We established experimental immune-mediated blepharoconjunctivitis (EC) in Brown Norway (BN) rats as a model for severe types of AC. We investigated whether macrophage infiltration in the Conjunctiva in this EC model is inhibited by clodronate liposomes (CL2MDP-lip). The numbers of ED1-positive but not ED2-positive macrophages in the Conjunctivas were increased by the induction of EC. SubConjunctival injection of CL2MDP-lip decreased the number of ED2-positive but not ED1-positive macrophages in the Conjunctivas of naive rats. CL2MDP-lip did not affect macrophages in the spleen. SubConjunctival injection of CL2MDP-lip into EC-developing BN rats decreased the number of ED2-positive macrophages at all the time points. ED1-positive cell infiltration was inhibited when treatment was administered just prior to OVA challenge. Intravenous injection of CL2MDP-lip decreased the number of ED2-positive cells in the Conjunctiva. Thus, we conclude that CL2MDP-lip inhibits infiltration of macrophages into the Conjunctiva within 24 h of antigen challenge.
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ag specific recognition activation and effector function of t cells in the Conjunctiva with experimental immune mediated blepharoconjunctivitis
Investigative Ophthalmology & Visual Science, 2003Co-Authors: Atsuki Fukushima, Akemi Ozaki, Waka Ishida, Kazuyo Fukata, Hisayuki UenoAbstract:PURPOSE. To investigate antigen (Ag) specificity, activation, and effector function of the Ag-specific T cells involved in the development of experimental immune-mediated blepharoconjunctivitis (EC), an experimental conjunctivitis. METHODS. EC was induced in Brown Norway rats by injection of ovalbumin (OVA)-specific T cells followed by OVA challenge with eye drops. Eyes, including the Conjunctivas, were harvested at different time points after challenge. The dependence of EC onset on the challenging Ag was assessed by challenge with an irrelevant Ag or stimulatory OVA peptides. To show the infiltration of transferred T cells into the Conjunctiva, T cells were labeled with 5-(and-6)-carboxyfluorescein diacetate succinimidyl ester (CFSE) before transfer. The activation of T cells in the Conjunctiva was assessed by measuring phosphorylation of Lck-associated molecules by Western blot analysis. Conjunctivas were also examined by immunohistochemistry and used for reverse transcription-polymerase chain reaction to determine the phenotype of the infiltrating cells and cytokine, chemokine, and chemokine receptor expression. To investigate infiltration of non Ag-specific T cells into the Conjunctiva, ragweed (RW)-primed lymphocytes were transferred into OVA-specific T-cell receptor transgenic (DOI1.10) mice. The mice were then challenged with RW and the Conjunctivas were harvested for immunohistochemistry to detect T cells derived from DO11.10 mice. RESULTS. EC was induced only when challenged with OVA protein or stimulatory OVA peptides, and CFSE-labeled transferred cells were found in the Conjunctiva. Phosphorylation of Lck and an 85-kDa Lck-associated molecule were observed in the Conjunctiva 6 hours after challenge. Many cytokines and chemokines began to be expressed at 6 hours, and individual expression patterns over time correlated well with the infiltration patterns of different inflammatory cells. In DO11.10 mice that received RW-primed lymphocytes, T cells derived from the recipient mice infiltrated the Conjunctiva after RW challenge. CONCLUSIONS. Ag-specific T cells initiate EC by first infiltrating the Conjunctiva, where they become activated by the specific Ag in the Conjunctiva.