g(m1)-ganglioside and Uveitis

Exogenous ganglioside GM1 has been reported to exert an immunomodulatory effect. We investigated the anti-inflammatory effect of GM1 ganglioside on endotoxin-induced uveitis (EIU) in rats and RAW 264.7 macrophages.. EIU was induced in Lewis rats by administering a subcutaneous injection of lipopolysaccharide (LPS). GM1 was injected intraperitoneally for three consecutive days prior to the LPS injection. Twenty-four hours after the LPS injection, the integrity of the blood-aqueous barrier was evaluated by determining the protein concentration and number of infiltrating cells in the aqueous humor (AqH). Immunohistochemical and Western blot analyses of the iris-ciliary body (ICB) were performed to evaluate the effect of GM1 on the LPS-induced expression of cyclooxygenase-2 (COX-2) and intercellular adhesion molecule-1 (ICAM-1). The effect of GM1 on proinflammatory mediators and signaling cascades was examined in LPS-stimulated RAW 264.7 cells using Western blotting and immunofluorescence staining to further clarify the underlying anti-inflammatory mechanism.. GM1 significantly reduced the protein concentration and number of infiltrating cells in the AqH of rats with EIU. GM1 also decreased the LPS-induced expression of the ICAM-1 and COX-2 proteins in the ICB. In RAW 264.7 cells, GM1 inhibited the proinflammatory mediators induced by LPS, including inducible nitric oxide synthase (iNOS), COX-2, tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6), and this inhibitory effect was potentially mediated by suppressing transforming growth factor-β-activated kinase 1 (TAK1) and reactive oxygen species (ROS)-mediated activation of nuclear factor-κB (NF-κB) and mitogen-activated protein kinases (MAPKs).. Based on this study, GM1 may be a potential anti-inflammatory agent for ocular inflammatory diseases.

Topics: Animals; Anti-Inflammatory Agents; Cyclooxygenase 2; G(M1) Ganglioside; Intercellular Adhesion Molecule-1; Interleukin-6; Lipopolysaccharides; Mice; Nitric Oxide; Rats; Rats, Inbred Lew; RAW 264.7 Cells; Uveitis

A potent cytolytic pore-forming protein (PFP, perforin, or cytolysin) is associated with the cytoplasmic granules of cytotoxic T lymphocytes (CTL) and natural killer (NK) cells. The role of PFP/perforin in cytolytic reactions carried out in vivo is still unclear. Here, the authors performed immunohistochemical analysis using antibodies monospecific for perforin and made use of a murine uveitis model produced by intracameral inoculation of herpes simplex virus I (HSV-I). The main cell infiltrate found in the anterior segment of virus-inoculated eyes consisted of Thy-1+/asialo GM1+/CD8-/CD4- cells, presumably representing NK cells. Perforin staining was detected mainly in cells bearing this phenotype. Perforin was only detected in cells displaying the large granular lymphocyte morphology. A small number of perforin-positive cells (less than 5%) colabeled as CD8+, indicating that these cells could have belonged to the CTL lineage. These observations show for the first time the presence of perforin-containing NK cells in tissues of animals undergoing acute viral infections.

Topics: Animals; Antibody Specificity; Fluorescent Antibody Technique; G(M1) Ganglioside; Glycosphingolipids; Herpes Simplex; Immune Sera; Killer Cells, Natural; Lymphocytes; Membrane Glycoproteins; Membrane Proteins; Mice; Mice, Inbred BALB C; Perforin; Pore Forming Cytotoxic Proteins; T-Lymphocytes, Cytotoxic; Uveitis; Virus Diseases

One of the most important components in the pathogenesis of lens opacification related to uveitis appears to be lens permeability changes consisting of potassium loss, sodium and water intake and late leakage of cytoplasmic constituents. These changes may be induced by 'Lens Permeability Factors' present and active in the aqueous humor during inflammation, factors that include antigen-antibody complexes, antilens and antiuveoretinal antibodies, phospholipase A and lysophosphatidylcholine (LPC). The damage caused by at least one of these factors, LPC, may be counteracted in vitro by a monosialoganglioside, (GM1). An in vivo experiment, still in progress, suggests that GM1 has a positive effect even in cases of incipient lens opacification related to uveitis.

Topics: Animals; Cataract; Cell Membrane Permeability; Cells, Cultured; Crystallins; G(M1) Ganglioside; Humans; Lens, Crystalline; Lysophosphatidylcholines; Phospholipases A; Rabbits; Uveitis