Biomedical subjects

Kazuyo Fukata

Publications and source records attributed to Kazuyo Fukata.

23 records · Page 2Linked to original sources

PubMed2002

Genetic background determines sensitivity to the inhibitory function of NO on T cell proliferation and the amounts of NO production mediated through IFN-gamma.

Abstract: Nitric oxide (NO) inhibits T cell proliferation. We demonstrate that the action of NO on T cell proliferation is different for Lewis and Brown Norway (BN) rats. Splenocytes from Lewis rats consistently showed higher proliferation against concanavalin A than splenocytes from BN rats did. In contrast, NO production was higher in BN rats than in Lewis rats. A depletion of adherent cells increased proliferation in BN rats to a level similar to that in Lewis rats. Thus NO produced by adherent splenocytes could be considered to inhibit proliferation. The addition of NG-monomethyl-L-arginine, a potent inhibitor of NO production, increased proliferation in Lewis rats, but much less so in BN rats. Similar results were obtained by the addition of anti-interferon (IFN)-gamma. It is surprising that, low doses of sodium nitroprusside, an NO donor, increased proliferation in BN rats but not in Lewis rats. To investigate the mechanism of differential NO production between the two strains, splenocytes were stimulated with IFN-gamma. The early signaling event evaluated by the phosphorylation of Stat-1 was similar in both strains, whereas inducible NO synthase (iNOS) mRNA expression seemed more sustained in BN rats. Thus the differential production of NO might be related to the differential transcriptional regulation of iNOS. Altogether, genetic background might be involved in sensitivity to the inhibitory function of NO for T cell proliferation and NO production.

Animals↗

PubMedDate not supplied

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

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.

Animals↗

PubMedDate not supplied

Suppression or exacerbation of experimental autoimmune uveoretinitis in Lewis rats by pretreatment with or without an autoantigenic peptide in aluminum hydroxide.

PURPOSE: Aluminum hydroxide (Al) is an adjuvant to induce Th2 immune responses. The aim of this study is to investigate the effects of pretreatment with Al on experimental autoimmune uveoretinitis (EAU), a Th1 disease model. METHODS: Lewis rats were pretreated with an uveitogenic peptide (#29)/Al, phosphate-buffered saline (PBS)/Al or PBS. EAU was then induced by immunization of #29 in complete Freund's adjuvant (CFA) with intravenous injection of Bordetella pertussis. Three weeks later, EAU was evaluated histologically and antigen-specific cellular immune responses were assessed. RESULTS: EAU was exacerbated in the PBS/Al group and attenuated in the #29/Al group, compared to the control PBS group. Antigen-specific cellular proliferation and interferon-gamma production were augmented in the PBS/Al group and suppressed in the #29/Al group. CONCLUSIONS: Suppression or exacerbation of EAU by the pretreatment in this study is related to inhibition or augmentation of antigen-specific Th1 immunity.

Adjuvants, Immunologic↗

PubMedDate not supplied

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

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.

Animals↗

PubMedDate not supplied

Cyclosporin A inhibits eosinophilic infiltration into the conjunctiva mediated by type IV allergic reactions.

BACKGROUND: Eosinophils are important effector cells in severe allergic conjunctivitis such as vernal keratoconjunctivitis. Infiltration of eosinophils into the conjunctiva is mediated by type I and type IV allergic reactions. Cyclosporin A (CsA) eye drops are administered therapeutically for severe allergic conjunctivitis, but the mechanism by which CsA acts, that is, by inhibiting type I, type IV or both types of allergic reactions, is not known. We investigated whether CsA eye drops inhibit type I, type IV or both types of allergic reactions in the conjunctiva. METHODS: Experimental immune-mediated blepharoconjunctivitis (EC) was induced in BALB/c mice by either active immunization or passive immunization by transfer of ragweed (RW)-primed splenocytes and RW-specific IgE, followed by RW challenge to the conjunctiva. These mice were treated in eye drops with vehicle, 0.1% CsA, 0.5% CsA or 0.1% betamethasone five times (1 and 2 h before RW challenge and 1, 2 and 3 h after RW challenge). Twenty-four hours after the challenge, the conjunctivas were harvested for histological analysis to evaluate eosinophilic infiltration. To evaluate effects of CsA eye drops on systemic immune responses, sera and spleens were collected from actively immunized mice at the time of sacrifice to examine serum IgE levels and cellular immune responses, respectively. RESULTS: CsA eye drops significantly inhibited eosinophilic infiltration into the conjunctiva in actively immunized EC-developing mice compared with vehicle-treated mice. The CsA-induced inhibition was similar to inhibition induced by 0.1% betamethasone. Serum IgE levels and splenocyte responses in CsA-treated mice were equivalent to those in vehicle-treated mice. Betamethasone treatment inhibited eosinophilic infiltration into the conjunctiva induced by both splenocyte transfer and IgE transfer, while CsA treatment inhibited infiltration induced by splenocyte transfer. CONCLUSIONS: CsA eye drops inhibited eosinophilic infiltration into the conjunctiva without affecting systemic immune responses. CsA predominantly inhibits eosinophilic infiltration by interfering with the type IV allergic reaction in the conjunctiva.

Animals↗