Skip header and navigation

Refine By

   MORE

6 records – page 1 of 1.

Ag-specific recognition, activation, and effector function of T cells in the conjunctiva with experimental immune-mediated blepharoconjunctivitis.

https://arctichealth.org/en/permalink/ahliterature50675
Source
Invest Ophthalmol Vis Sci. 2003 Oct;44(10):4366-74
Publication Type
Article
Date
Oct-2003
Author
Atsuki Fukushima
Akemi Ozaki
Kazuyo Fukata
Waka Ishida
Hisayuki Ueno
Author Affiliation
Department of Ophthalmology, Kochi Medical School, Nankoku City, Japan. fukusima@kochi-ms.ac.jp
Source
Invest Ophthalmol Vis Sci. 2003 Oct;44(10):4366-74
Date
Oct-2003
Language
English
Publication Type
Article
Keywords
Animals
Blepharitis - immunology
Blotting, Western
Chemokines - metabolism
Conjunctiva - immunology
Conjunctivitis - immunology
Cytokines - metabolism
Epitopes - immunology
Flow Cytometry
Fluoresceins
Immunoenzyme Techniques
Immunophenotyping
Lymphocyte Activation - physiology
Male
Mice
Mice, Inbred BALB C
Ovalbumin - immunology
Rats
Rats, Inbred BN
Receptors, Chemokine - metabolism
Research Support, Non-U.S. Gov't
Reverse Transcriptase Polymerase Chain Reaction
Succinimides
T-Lymphocytes - physiology
Abstract
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 (DO11.10) 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.
PubMed ID
14507881 View in PubMed
Less detail

Analysis of Ag-presenting cells in the conjunctiva during the development of experimental immune-mediated blepharoconjunctivitis.

https://arctichealth.org/en/permalink/ahliterature50604
Source
Curr Eye Res. 2004 Oct-Nov;29(4-5):277-86
Publication Type
Article
Author
Akemi Ozaki
Atsuki Fukushima
Waka Ishida
Olga Zinchuk
Kazuyo Fukata
Yoshihiro Hayashi
Hideaki Enzan
Masaru Takata
Hideo Yagita
Hisayuki Ueno
Author Affiliation
Department of Ophthalmology, Kochi Medical School, Kohasu, Oko-cho, Nankoku City 783-8505, Japan.
Source
Curr Eye Res. 2004 Oct-Nov;29(4-5):277-86
Language
English
Publication Type
Article
Keywords
Animals
Antigen-Presenting Cells - immunology
Antigens, CD - metabolism
Antigens, CD86
Blepharitis - immunology
Blotting, Western
Cell Count
Conjunctiva - immunology
Conjunctivitis, Allergic - immunology
Extracellular Signal-Regulated MAP Kinases - metabolism
Flow Cytometry
Histocompatibility Antigens Class II - metabolism
Immunoenzyme Techniques
Immunophenotyping
Lymphocyte Activation
Macrophages - immunology
Male
Membrane Glycoproteins - metabolism
Microscopy, Confocal
Models, Theoretical
Ovalbumin - immunology
Phosphorylation
Rats
Rats, Inbred BN
T-Lymphocytes - immunology
Abstract
PURPOSE: The aim of this study was to investigate the phenotypes of antigen (Ag) presenting cells (APCs) in the conjunctiva during the development of experimental immune-mediated blepharoconjunctivitis (EC), which serves as a model for investigation of severe types of human allergic conjunctivitis. METHODS: Brown Norway rats treated by ovalbumin (OVA) were used in this study. To confirm the restriction of MHC class II by OVA-specific T cells, monoclonal Abs against MHC class II were added to the conventional proliferation assay. To evaluate the MHC class II expression in the conjunctiva during the development of EC, an immunohistochemical analysis, either as the single or double staining, was performed. Conjunctival fibroblast cell lines were established from naive rats and the MHC class II expression was evaluated by flow cytometric analysis. To examine the roles of costimulatory molecules, OVA-specific T cells were stimulated with anti-TcR Ab and anti-CD28 Ab and then subjected for Western blotting to evaluate the ERK phosphorylation. Finally, in vivo expression of B7 molecules was examined immunohistochemically. RESULTS: OVA-specific T cells recognized OVA in the context of MHC class II. MHC class II was expressed in conjunctival macrophages but not in fibroblasts. EC induction was accompanied by abundant infiltration of macrophages positive for MHC class II. MHC class II was also expressed in conjunctival epithelial cells by EC induction. Stimulation from CD28 was necessary for ERK phosphorylation. B7-2, but not B7-1, was expressed in the conjunctiva. CONCLUSION: Conjunctival macrophages may represent a major source of APCs for the induction of EC in the conjunctiva.
PubMed ID
15590473 View in PubMed
Less detail

Differential expression and signaling of IFN-gamma in the conjunctiva between Lewis and Brown Norway rats.

https://arctichealth.org/en/permalink/ahliterature50668
Source
Microbiol Immunol. 2003;47(10):785-96
Publication Type
Article
Date
2003
Author
Atsuki Fukushima
Akemi Ozaki
Kazuyo Fukata
Hisayuki Ueno
Author Affiliation
Laboratory of Immunology, Department of Ophthalmology, Kochi Medical School, Nankoku, Kochi 783-8505, Japan. fukusima@kochi-ms.ac.jp
Source
Microbiol Immunol. 2003;47(10):785-96
Date
2003
Language
English
Publication Type
Article
Keywords
Animals
Blepharitis - immunology
CD4-Positive T-Lymphocytes - immunology
Cell Line
Conjunctiva - immunology
Conjunctivitis, Allergic - immunology
DNA-Binding Proteins - metabolism
Humans
Interferon Type II - immunology - metabolism
Male
Ovalbumin - immunology
Phosphorylation
Rats
Rats, Inbred BN
Rats, Inbred Lew
Receptors, Interferon - metabolism
Research Support, Non-U.S. Gov't
STAT1 Transcription Factor
Signal Transduction
Species Specificity
Trans-Activators - metabolism
Abstract
Genetic background determines the histological features of experimental immune-mediated blepharoconjunctivitis (EC) in rats, which is a model for human allergic conjunctivitis (AC). A great number of lymphocytes predominate in EC of Lewis rats, while less lymphocytes and more eosinophils are present in that of Brown Norway (BN) rats. Although this difference could be attributed to their systemic Th1/Th2 dominancy, it remains unclear whether some regulatory mechanisms may exist in the inflammatory site in the conjunctiva. Here, we aim to investigate this hypothesis by comparing the expression levels of inflammatory mediators in the conjunctiva between the two strains. EC was induced in Lewis and BN rats by transfer of ovalbumin (OVA)-specific CD4(+) T-cell lines followed by eye drops of OVA as antigen challenge, and then was clinically and histologically evaluated. Reverse-transcription (RT)-PCR was performed to compare the expressions of cytokines and cytokine receptors (Rs) in conjunctivas of both strains of rats either with or without EC. To confirm the biological significance of interferon (IFN)-gamma R expression, phosphorylation of signal transducers and activators of transcription (STAT)-1 was examined in the conjunctivas, followed by subconjunctival injection of IFN-gamma. BN T cells contained interleukin (IL)-4 and IFN-gamma, while Lewis T cells expressed no IL-4. Transfer of those cells induced more severe EC in Lewis rats. RTPCR using naive conjunctivas detected more IL-4, IFN-gamma, and IFN-gamma R beta-chain RNA expression in BN rats. After the EC induction, BN rats expressed significantly higher amounts of IFN-gamma R beta-chain, and upregulation of interferon regulatory factor (IRF)-1 was observed. Phosphorylation of STAT-1 was more remarkable in BN rats. The findings demonstrate differential expression of IFN-gamma R and signaling through IFN-gamma in the conjunctiva between the two strains. This may be due to differences in histopathological character between the two strains.
PubMed ID
14605445 View in PubMed
Less detail

Mast-cell activation augments the late phase reaction in experimental immune-mediated blepharoconjunctivitis.

https://arctichealth.org/en/permalink/ahliterature50704
Source
Graefes Arch Clin Exp Ophthalmol. 2003 May;241(5):394-402
Publication Type
Article
Date
May-2003
Author
Akemi Ozaki
Atsuki Fukushima
Kazuyo Fukata
Hisayuki Ueno
Author Affiliation
Laboratory of Immunology, Department of Ophthalmology, Kochi Medical School, Kohasu, Oko-cho, 783-8505 Nankoku-city, Japan.
Source
Graefes Arch Clin Exp Ophthalmol. 2003 May;241(5):394-402
Date
May-2003
Language
English
Publication Type
Article
Keywords
Animals
Blepharitis - chemically induced - immunology - pathology
Cell Movement
Conjunctivitis, Allergic - chemically induced - immunology - pathology
Enzyme-Linked Immunosorbent Assay
Eosinophilia - immunology
Eosinophils - physiology
Flow Cytometry
Hypersensitivity, Delayed - immunology - pathology
Lymphocyte Activation
Male
Mast Cells - physiology
Ovalbumin - immunology
RANTES - metabolism
Rats
Rats, Inbred BN
Research Support, Non-U.S. Gov't
Reverse Transcriptase Polymerase Chain Reaction
T-Lymphocytes - immunology
p-Methoxy-N-methylphenethylamine
Abstract
BACKGROUND: How the early phase allergic reaction affects the late phase reaction remains unclear. We examined this issue with an experimental model of allergic conjunctivitis that permits the two reactions to be disconnected from each other. METHODS: Experimental immune-mediated blepharoconjunctivitis (EC) was initiated in Brown Norway rats by transferring ovalbumin (OVA)-specific T cells and then challenging with OVA-containing eye drops. To induce early phase reaction, a mast-cell activator, C48/80, was challenged together with or without OVA. Rats were evaluated clinically and eyes were harvested for histologic examination and for evaluation of chemokine expression by reverse-transcriptase PCR. RESULTS: The rats challenged with OVA alone developed the T-cell-mediated late phase reaction histologically, but not clinically, in the absence of early phase reaction. While rats challenged with C48/80 with or without OVA exhibited clinical signs of the early phase reaction, the clinical late phase reaction was observed only in the OVA+C48/80 group. Eosinophilic infiltration into the conjunctiva during the late phase reaction of the OVA+C48/80 group markedly exceeded that of rats challenged with either OVA or C48/80 alone. RANTES (regulated on activation, normal T-cell expressed and secreted), an eosinophil attractant, was expressed both in the OVA+C48/80 and OVA groups, while eotaxin was expressed at equivalent levels in all three groups. CONCLUSION: The mast-cell-mediated early phase reaction potentiates the T-cell-mediated late phase reaction, and RANTES is involved in eosinophilic infiltration induced by antigen-specific T cells. Other molecules induced by allergen-specific T cells activated in an as yet unknown manner by the mast cells may be responsible for the infiltration of eosinophils.
PubMed ID
12682842 View in PubMed
Less detail

Phenotypic changes and inflammatory cell distribution in the cornea during development of experimental immune-mediated blepharoconjunctivitis.

https://arctichealth.org/en/permalink/ahliterature50616
Source
Jpn J Ophthalmol. 2004 Jul-Aug;48(4):333-9
Publication Type
Article
Author
Akemi Ozaki
Waka Ishida
Kazuyo Fukata
Atsuki Fukushima
Hisayuki Ueno
Author Affiliation
Laboratory of Immunology, Department of Ophthalmology, Kochi Medical School, Nankoku, Japan.
Source
Jpn J Ophthalmol. 2004 Jul-Aug;48(4):333-9
Language
English
Publication Type
Article
Keywords
Animals
Antigens, CD - metabolism
Blepharitis - immunology - pathology
Conjunctivitis, Allergic - immunology - pathology
Cornea - pathology
Disease Models, Animal
Eosinophils - pathology
Immunoenzyme Techniques
Macrophages - pathology
Male
Ovalbumin - immunology
Phenotype
Rats
Rats, Inbred BN
T-Lymphocytes - pathology
Abstract
PURPOSE: To investigate the morphological changes in the cornea during the development of experimental immune-mediated blepharoconjunctivitis (EC). METHODS: EC was induced in Brown Norway (BN) rats by active immunization with ovalbumin (OVA) emulsified in complete Freund's adjuvant and a subsequent challenge by OVA eyedrops. The corneas were analyzed immunohistochemically. RESULTS: Before the induction of EC, cells stained with OX6 (rat MHC class 2, RT1B), ED1 (tissue macrophages), ED2 (resident macrophages), CD4, or major basic protein were present in the peripheral corneal stroma. ED1- and OX6-stained cells were also observed in the central corneal stroma, and their number increased after the antigen challenge. Infiltration of cells stained with ED1, ED2, OX62 (dendritic cells), CD4, or CD3 (T cells) from the limbus to the peripheral corneal stroma started 6 h after the antigen challenge. Expression of MHC class 2 molecules was induced on the corneal epithelium by the antigen challenge. CONCLUSIONS: The present study demonstrates for the first time the phenotypic changes and distribution of inflammatory cells in the cornea during the development of EC.
PubMed ID
15295657 View in PubMed
Less detail

Suppression of experimental immune-mediated blepharoconjunctivitis in Brown Norway rats by topical application of FK506.

https://arctichealth.org/en/permalink/ahliterature57458
Source
Graefes Arch Clin Exp Ophthalmol. 2002 Feb;240(2):137-43
Publication Type
Article
Date
Feb-2002
Author
Koji Nishino
Atsuki Fukushima
Shigeki Okamoto
Yuichi Ohashi
Kazuyo Fukata
Akemi Ozaki
Hiasyuki Ueno
Author Affiliation
Department of Ophthalmology, Kochi Medical School, Nankoku, Japan.
Source
Graefes Arch Clin Exp Ophthalmol. 2002 Feb;240(2):137-43
Date
Feb-2002
Language
English
Publication Type
Article
Keywords
Administration, Topical
Animals
Blepharitis - drug therapy - immunology - pathology
Conjunctivitis - drug therapy - immunology - pathology
Immunization, Passive
Immunosuppressive Agents - therapeutic use
Male
Ophthalmic Solutions
Ovalbumin - immunology
Rats
Rats, Inbred BN
Research Support, Non-U.S. Gov't
Tacrolimus - administration & dosage - therapeutic use
Abstract
BACKGROUND: Experimental immune-mediated blepharoconjunctivitis (EC) in Brown Norway (BN) rats, which is inducible by transfer of antigen-specific T cells, is a model for human allergic conjunctivitis. We investigated the possible inhibition of EC in BN rats by topical application of FK506, which is an immunosuppressive agent that mainly targets T cells. METHODS: To induce EC by active immunization, ovalbumin (OVA) adsorbed to alum was injected into the hind footpads of BN rats. Three weeks after the initial immunization, rats were challenged with OVA by eye drops. Twenty-four hours later, lids including conjunctivas, lymph nodes (LNs), and sera were harvested for histology or reverse transcriptase PCR, proliferation assays, and measurement of IgE titer, respectively. For passive immunization, rats were intravenously injected with 10 million of in vitro-stimulated OVA-primed LN cells. Four days after the transfer, rats were challenged with OVA and evaluated as above. The rats were divided into two groups. One group received topical FK506 treatment three times per day from 15 to 21 days after active immunization or from 1 to 4 days after transfer. The other group was treated with vehicle as above. RESULTS: FK506 treatment suppressed infiltration of both lymphocytes and eosinophils in the conjunctiva either by active or passive immunization (P
PubMed ID
11931080 View in PubMed
Less detail

6 records – page 1 of 1.