i(3)so3-galactosylceramide and Inflammation

Topics: alpha 1-Antitrypsin; alpha-Macroglobulins; Animals; Antigen-Antibody Complex; Antithrombin III; Basement Membrane; Blood Coagulation; Blood Proteins; Cattle; Collagen; Complement C1 Inactivator Proteins; Factor XI; Factor XII; Humans; Inflammation; Kininogens; Leukocytes; Lipopolysaccharides; Partial Thromboplastin Time; Prekallikrein; Protein Binding; Rabbits; Sulfoglycosphingolipids; Uric Acid

Sulfatide is a sulfated glycosphingolipid that is present abundantly in myelin sheaths of the brain and spinal cord. It is synthesized by a cerebroside sulfotransferase encoded by Gal3st1, which catalyzes the transfer of sulfate from 3'-phosphoadenylylsulfate to galactosylceramide. We previously reported that Gal3st1 gene expression in the spinal cord is up-regulated 1 day after intraplantar injection of complete Freund's adjuvant (CFA), indicating that sulfatide is involved in inflammatory pain. In the present study, we found that intrathecal injection of sulfatide led to mechanical allodynia. Sulfatide caused levels of glial fibrillary acidic protein (GFAP) and nitric oxide in the spinal cord to increase. Mechanical allodynia induced by intrathecal injection of sulfatide was blocked by nitric oxide synthase inhibitors and by suppression of astrocyte activation by L-α-aminoadipate. These results suggest that sulfatide-induced mechanical allodynia involved glial activation and nitric oxide production. Blocking selectin, a sulfatide-binding protein, with bimosiamose attenuated sulfatide-induced allodynia and ameliorated CFA-induced mechanical allodynia during inflammatory pain. Finally, elevated levels of sulfatide concentration in the spinal cord were observed during CFA-induced inflammatory pain. The elevated sulfatide levels enhanced selectin activation in the spinal cord, resulting in mechanical allodynia. Our data suggest that sulfatide-selectin interaction plays a key role in inflammatory pain.

Topics: Humans; Hyperalgesia; Inflammation; Nitric Oxide; Pain; Spinal Cord; Sulfoglycosphingolipids

CD1d-restricted NKT cells can be divided into two groups: type I NKT cells use a semi-invariant TCR, whereas type II express a relatively diverse set of TCRs. A major subset of type II NKT cells recognizes myelin-derived sulfatides and is selectively enriched in the CNS tissue during experimental autoimmune encephalomyelitis (EAE). We have shown that activation of sulfatide-reactive type II NKT cells by sulfatide prevents induction of EAE. In this article, we have addressed the mechanism of regulation, as well as whether a single immunodominant form of synthetic sulfatide can treat ongoing chronic and relapsing EAE in SJL/J mice. We have shown that the activation of sulfatide-reactive type II NKT cells leads to a significant reduction in the frequency and effector function of myelin proteolipid proteins 139-151/I-A(s)-tetramer(+) cells in lymphoid and CNS tissues. In addition, type I NKT cells and dendritic cells (DCs) in the periphery, as well as CNS-resident microglia, are inactivated after sulfatide administration, and mice deficient in type I NKT cells are not protected from disease. Moreover, tolerized DCs from sulfatide-treated animals can adoptively transfer protection into naive mice. Treatment of SJL/J mice with a synthetic cis-tetracosenoyl sulfatide, but not α-galactosylceramide, reverses ongoing chronic and relapsing EAE. Our data highlight a novel immune-regulatory pathway involving NKT subset interactions leading to inactivation of type I NKT cells, DCs, and microglial cells in suppression of autoimmunity. Because CD1 molecules are nonpolymorphic, the sulfatide-mediated immune-regulatory pathway can be targeted for development of non-HLA-dependent therapeutic approaches to T cell-mediated autoimmune diseases.

Topics: Adoptive Transfer; Animals; Cattle; CD4-Positive T-Lymphocytes; Clonal Anergy; Dendritic Cells; Encephalomyelitis, Autoimmune, Experimental; Female; Humans; Inflammation; Mice; Mice, Inbred C57BL; Mice, Inbred Strains; Multiple Sclerosis, Relapsing-Remitting; Myelin Proteolipid Protein; Natural Killer T-Cells; Sulfoglycosphingolipids

To develop more effective vaccines and strategies to regulate chronic inflammatory diseases, it is important to understand the mechanisms of immunological memory. Factors regulating memory CD4(+) T helper (Th)-cell pool size and function remain unclear, however. We show that activation of type I invariant natural killer T (iNKT) cells with glycolipid ligands and activation of type II natural killer T (NKT) cells with the endogenous ligand sulfatide induced dramatic proliferation and expansion of memory, but not naïve, CD4 T cells. NKT cell-induced proliferation of memory Th1 and Th2 cells was dependent largely on the production of IL-2, with Th2-cell proliferation also affected by loss of IL-4. Type II NKT cells were also required for efficient maintenance of memory CD4 T cells in vivo. Activation of iNKT cells resulted in up-regulation of IFN-γ expression by memory Th2 cells. These IFN-γ-producing memory Th2 cells showed a decreased capability to induce Th2 cytokines and eosinophilic airway inflammation. Thus, activated NKT cells directly regulate memory CD4 T-cell pool size and function via the production of cytokines in vivo.

Topics: Animals; Antigens, CD1d; CD4-Positive T-Lymphocytes; Cell Proliferation; Glycolipids; Immunologic Memory; Inflammation; Interferon-gamma; Interleukin-2; Interleukin-4; Killer Cells, Natural; Mice; Mice, Knockout; Sulfoglycosphingolipids; Th2 Cells

Sulfatides are known to be a native ligand of P-selectin. Platelet-leukocyte interaction via the cross-talk between P-selectin and Mac-1 (CD11b/CD18) plays an important role in the mechanism of neointimal thickening after vascular injury such as that seen in post-stent restenosis. However, the roles of sulfatides on restenosis have not been elucidated.. Serum sulfatide levels, P-selectin expression on the surface of platelets, and activated Mac-1 on the surface of neutrophils were serially measured using both coronary sinus and peripheral blood samples in 21 patients who underwent coronary stent implantation.. The trans-cardiac gradient (coronary sinus minus peripheral blood) of the sulfatide levels significantly increased at 15 min (-1.47+/-2.87 to 0.59+/-1.44 nmol/ml, p<0.001), compared to baseline levels. The maximum response of the trans-cardiac gradient of P-selectin expression on the surface of platelets at 15 min after stent implantation (R=0.55, p<0.01), and that of activated Mac-1 on the surface of neutrophils at 48 h (R=0.59, p<0.01), were both positively correlated with that of serum sulfatide levels at 15 min. The angiographic late lumen loss was correlated with the trans-cardiac gradient of sulfatide levels at 15 min (R=0.48, p<0.05), platelet P-selectin expression at 15 min (R=0.42, p<0.05) and activated neutrophil Mac-1 expression at 48 h (R=0.46, p<0.05), but not with values at other sampling points.. Sulfatides may play a physiological role on inflammation in vascular injury and the development of neointimal thickening.

Topics: Aged; Angioplasty, Balloon, Coronary; Female; Humans; Inflammation; Macrophage-1 Antigen; Male; Middle Aged; P-Selectin; Stents; Sulfoglycosphingolipids; Tunica Intima

Sulfatide, a major lipid component of myelin sheath, participates in diverse cellular events of the CNS, and its cellular level has recently been implicated in many inflammation-associated neuronal diseases. Herein, we report that sulfatide alone can trigger pathological inflammatory responses in glia, brain-resident immune cells. We show that sulfatide changed the morphology of primary microglia to their activated form, and it significantly induced the production of various inflammatory mediators in primary microglia and astrocytes. Moreover, sulfatide rapidly triggered the phosphorylation of p38, ERK, and JNK within 30 min, and it markedly enhanced the NF binding activity to NF-kappaB and AP-1 binding elements. However, nonsulfated galactocerebroside, another major lipid component of myelin, had no effect on activation of glia. We further reveal that CD1d did not contribute to sulfatide-stimulated activation of MAPKs, although its expression was enhanced by sulfatide and sulfatide-treated microglial cells actually stimulated type II NKT cells. Sulfatide significantly stimulated the phosphorylation of MAPKs in glia from CD1d-deficient mice, and the phosphorylation levels were similar to those in wild-type littermates. Sulfatide-triggered inflammatory events appear to occur at least in part through an L-selectin-dependent mechanism. L-selectin was dramatically down-regulated upon exposure to sulfatide, and inhibition of L-selectin resulted in suppression of sulfatide-triggered responses. Collectively, these results show that abnormally released sulfatide at demyelinated regions may act as an endogenous stimulator in the brain immune system, thus causing and further exacerbating pathological conditions in the brain.

Topics: Animals; Antigens, CD1d; Brain Diseases; Down-Regulation; Enzyme Activation; Extracellular Signal-Regulated MAP Kinases; Inflammation; Inflammation Mediators; L-Selectin; MAP Kinase Kinase 4; Mice; Microglia; Myelin Sheath; Natural Killer T-Cells; NF-kappa B; p38 Mitogen-Activated Protein Kinases; Sulfoglycosphingolipids; Time Factors; Transcription Factor AP-1

Sulfatides highly expressed in the normal gastric mucosa play important roles in gastric mucosal protection. However, it is unknown whether the sulfatides expression changes in chronic gastritis.. Sulfatides expression levels were examined with immunohistochemical staining in 77 specimens obtained from resected human stomachs, followed by the measurement of optical density of the staining under digital microscopy. For mRNA expression analysis of GalCer (galactosylceramide)-sulfotransferase and arylsulfatase A using a quantitative real time reverse transcription-polymerase chain reaction, 64 biopsy specimens were endoscopically taken from the gastric corpus of out patients. Mucosal changes were scored under microscopic observations according to the updated Sydney System classification.. The sulfatides expression decreased along with the progression of mucosal atrophy and lymphocytes infiltration, and was barely observed in intestinal metaplasia. The mRNA expression of GalCer-sulfotransferase, a biosynthetic enzyme for sulfatides, also decreased along with the progression of mucosal atrophy and lymphocytes infiltration. In contrast, mRNA expression of arylsulfatase A, which degrades sulfatides, was not altered.. Expression of sulfatides and GalCer-sulfotransferase in the gastric mucosa of chronic gastritis is selectively reduced according to the mucosal atrophy and inflammation. Decreased expression of sulfatides may attribute to the deterioration of mucosal protection in chronic gastritis.

Topics: Atrophy; Biopsy; Cerebroside-Sulfatase; Chronic Disease; Down-Regulation; Gastric Mucosa; Gastritis; Humans; Immunohistochemistry; Inflammation; Lymphocytes; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Sulfoglycosphingolipids; Sulfotransferases

There is increasing evidence that the ligation of adhesion molecules such as L-selectin can activate phagocytes to their full inflammatory potential. Sulfatide has been established as ligand for L-selectin and shown to trigger intracellular signals in human neutrophils. However, it remains unclear whether the ligation of L-selectin with sulfatide affects neutrophil phagocytosis. We studied the effects of sulfatide upon Fc gamma R- and CR3-mediated human neutrophil phagocytosis. Adhesion of the cells to a sulfatide-coated surface resulted in a dose-dependent enhancement of phagocytosis mediated via Fc gamma R or CR3, or both receptors. Galactocerebroside, but not glucocerebroside, also enhanced phagocytosis by neutrophils; therefore, galactose residue is thought to be required on ceramide molecules for the activation. Chymotrypsin-treated neutrophils, from which most L-selectin had been removed, reacted with sulfatide and galactocerebroside to enhance phagocytosis. These results suggest that an unidentified receptor for these cerebrosides exists on neutrophils and participates in the enhancement of phagocytosis.

Topics: Cell Adhesion; Cell Membrane; Cell Separation; Cells, Cultured; Ceramides; Cerebrosides; Chymotrypsin; Dose-Response Relationship, Drug; Flow Cytometry; Galactosylceramides; Glucosylceramides; Humans; Inflammation; L-Selectin; Macrophage-1 Antigen; Neutrophils; Phagocytes; Phagocytosis; Receptors, IgG; Sulfoglycosphingolipids