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PubMed1987

Interleukin 3-dependent mouse mast cells express the cholera toxin-binding acidic glycosphingolipid, ganglioside GM1, and increase their histamine content in response to toxin.

The acidic glycosphingolipid, ganglioside GM1, which is the binding site for cholera toxin on many cell types, was identified by chemical and by flow cytometric analyses of mouse interleukin 3-dependent, bone marrow culture-derived mast cells (BMMC). Ganglioside GM1 and other acidic glycosphingolipids were isolated from BMMC by chloroform/methanol extraction and chromatography on DEAE-Sephadex and were analyzed by thin layer chromatography. The presence of ganglioside GM1 in the BMMC extract was demonstrated by its co-migration with ganglioside GM1 standard in thin layer chromatography and by the binding of peroxidase-labeled cholera toxin B subunit to both molecules. As assessed by fluorescence flow cytometric analysis of the binding of fluorescein-conjugated cholera toxin B subunit, the majority of BMMC expressed ganglioside GM1 on their surface, and the total presentation per cell increased as cells progressed from the G1 to S to G2 + M phases of the cell cycle. The addition of increasing amounts of cholera toxin starting with 0.08 microgram/ml to BMMC cultured in 50% WEHI 3-conditioned medium containing IL 3 for 48 hr caused the adhesion of BMMC to the tissue culture flasks to increase in a dose-related manner, from less than 1% adherent cells in cultures without toxin to a plateau value of approximately 17% adherent in the presence of 1.25 micrograms/ml of toxin. The histamine content of BMMC increased from 26.7 +/- 3.59 ng/10(6) cells (mean +/- SD, n = 4) for control cultures to 201 +/- 17.4 ng/10(6) cells (mean +/- SD, n = 4) for nonadherent cells and to 588 +/- 89.4 ng/10(6) cells (mean +/- SD, n = 4) for adherent cells after 48 hr of culture in 0.31 microgram/ml cholera toxin, which was the optimal dose for nonadherent and adherent populations. The content of another preformed intragranular mediator, beta-hexosaminidase, did not increase appreciably in the presence of cholera toxin (n = 3). The increase in the histamine content of BMMC after the addition of 0.31 microgram/ml cholera toxin was detectable at 4 hr, plateaued by 24 to 48 hr, and gradually declined over the next 6 days. Cholera toxin also augmented the histamine content of BMMC in the presence of purified synthetic IL 3. Preincubation of whole cholera toxin with purified ganglioside GM1 inhibited the histamine-augmenting effects of cholera toxin on BMMC, indicating that the effect was not due to a contaminant, and neither the A nor B subunit of cholera toxin alone increased the histamine content of BMMC.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1988

Effect of gangliosides on primary and secondary plaque-forming cells responses and on suppressor T-cells induction.

This study was designed to determine the suppressive effect of bovine brain gangliosides on primary and secondary anti-sheep red blood cell (SRBC) plaque-forming cells (PFC) response. Spleen cells from inbred C57BL/6J mice were used as sources of lymphocytes. The result showed that levels of gangliosides at 50 and 100 micrograms/ml suppressed primary anti-SRBC PFC responses about 30% and 60%, respectively, of normal control. In contrast, secondary anti-SRBC PFC were not suppressed by the same levels of gangliosides. Likewise, PFC response from mixing SRBC-primed T-cells with naive B-cells was not suppressed by gangliosides. In addition, induction of suppressor T-cells was reduced by 50 and 100 micrograms/ml of gangliosides, 57% and 63%, respectively, versus 86% of suppression of PFC response by the control suppressor T-cells. These results further suggest the role of gangliosides as immunomodulators.

Animals↗
PubMed1985

Gangliosides in early interactions between vesicular stomatitis virus and CER cells.

In the present report an attempt was made to elucidate the role of gangliosides in early interactions between vesicular stomatitis virus (VSV) and CER cells. Research was carried out to test the ability of gangliosides from mammal brains and from CER cells to inhibit viral attachment to susceptible cells. The incubation of VSV in the presence of gangliosides decreased the subsequent infection of CER cells by the virus. When similar experiments were performed with gangliosides inserted in liposomes the inhibition of infection was enhanced. Since carbohydrate moieties could participate to rhabdovirus binding as a part of a glycolipid receptor, CER cells were subjected to the action of glycosidases and these produced a fall in the viral attachment. Deglycosilated CER cells reacquired their susceptibility to virus infection after coating with gangliosides immediately after enzyme treatment. Results obtained show the participation of gangliosides in the receptorial structure for vesiculovirus of susceptible CER cells.

Animals↗
PubMedDate not supplied

[Concentration and composition of gangliosides in portions of the vertebrate brain].

Studies have been made on gangliosides from various parts of the brain in 5 species of cartilaginous fishes, 2 species of teleosts, 2 amphibian species, 2 avian species and 3 species of mammals. Almost in all the species investigated, the highest content of gangliosides per weight unit of the brain were found in the forebrain (determinations were carried out by sialic acid, sometimes by sphingosine). Ganglioside content of brain structure in warm-blooded animals is higher that in corresponding structures of fishes and amphibia. Comparative studies on ganglioside composition in the brain of fishes and mammals indicate that brain structures exhibit the same pattern of molecular organization, as the whole brain of the species studied (high polysialoganglioside content in teleosts, predominance of gangliosides with a short carbon chain in cartilaginous fishes, etc.). Besides this fact, peculiarities of composition typical of the brain structures were found as well (more polar composition of gangliosides of the cerebellum and some other ones).

Animals↗
PubMed1988

Analysis of the expression of N-glycolylneuraminic acid-containing gangliosides in cells and tissues using two human monoclonal antibodies.

The specificities of two human monoclonal antibodies (2-39M and 32-27M), produced by hybridomas derived from the lymphocytes of melanoma patients (Yamaguchi, H., Furukawa, K., Fortunato, S. R., Livingston, P. O., Lloyd, K. O., Oettgen, H. F., and Old, L. J. (1987) Proc. Natl. Acad. Sci. U.S.A. 84, 2416-2420) have been elucidated. Using a large panel of glycolipids, it has been shown that the two monoclonal antibodies (mAbs) identified a number of N-glycolylneuraminic acid (NeuGc)-containing gangliosides. mAb 2-39M reacted with (NeuGc)GM3, (NeuGc)sialylparagloboside, and (NeuGc)sialylhexaglycosylceramide; no reactivity was observed with gangliosides containing only N-acetylneuraminic acid (NeuAc) or with disialogangliosides. These reactive species have the NeuGc alpha 2----3Gal- sequence in common. mAb 32-27M reacted strongly with (NeuGc)2 GD3 and (NeuGc)2disialylparagloboside, and moderately with (NeuAc-NeuGc-)GD3 and (NeuAc-NeuGc-)disialylparagloboside. The reactive species have sialic acid alpha 2----8NeuGc alpha 2----3Gal- sequences in common. These two antibodies were used to demonstrate the species-related presence of different NeuGc-containing gangliosides in various animal erythrocytes by thin layer chromatography immunostaining. No reactivity of either mAb was observed with gangliosides isolated from fresh human colon cancer, melanoma specimens, or some normal tissues, including brain. On the other hand, it was shown that mAb 32-27M reacted with gangliosides isolated from human melanoma and astrocytoma cells grown in fetal bovine serum but not from those grown in synthetic medium. Within the sensitivities of the methods used, these data, and related chemical analyses, do not support the presence of NeuGc-containing gangliosides in human tumors.

Animals↗
PubMedDate not supplied

[The content and composition of gangliosides in brain subcellular fractions of the frog Rana temporaria].

Ganglioside distribution in various frog brain subcellular fractions (myelin, microsomes, mitochondria, synaptosomes, plasma membranes of nerve endings and synaptic vesicles) was investigated. The synaptosomes and plasma membranes of nerve endings were found to be the main places of ganglioside localization, ganglioside concentration being 2.42 and 1.79 times higher than that in homogenates. Gangliosides were shown to be present in synaptic vesicles. The characteristic features of gangliosides from frog brain and its subcellular fractions are the predominance of polysialogangliosides with 3-5 sialic acid residues (up to 57.4%), low content of monosialogangliosides (not more than 7%) and the presence of disialogangliosides with short carbohydrate chain. The increase of ganglioside content per one nerve cell during phylogenetic development of vertebrates is discussed.

Animals↗
PubMed1988

Complex ganglioside expression and tetanus toxin binding by PC12 pheochromocytoma cells.

Ganglioside expression and tetanus toxin binding were studied in the rat pheochromocytoma cell line PC12. Seven ganglioside species were readily detected in extracts of PC12 cells; two were identified as tri- and tetrasialogangliosides, which are common brain constituents but unusual components of neuronal cell lines. Carbohydrate composition, acid and enzyme hydrolyses, and mass spectral analysis revealed that the major species is GT 1b, a predominant mammalian brain ganglioside previously reported to support high affinity tetanus toxin binding (Rogers, T. B., and Snyder, S. H. (1981) J. Biol. Chem. 256, 2402-2407). Direct binding of 125I-tetanus toxin to PC12 gangliosides on TLC plates revealed selective binding to the tri- and tetrasialogangliosides. Radioiodinated toxin also bound with high affinity to intact PC12 cells or their isolated membranes. The binding affinity (Kd = 1.25 nM), density of receptors (Bmax = 238 pmol/mg of membrane protein), and dependence on pH, ionic strength, and temperature were similar to those previously reported for toxin binding to rat brain synaptic membranes. Differentiation of PC12 cells caused an increase in expression of the tri- and tetrasialogangliosides and a closely matched increase in tetanus toxin binding to cell membranes. These data provide evidence that complex gangliosides may act as tetanus toxin receptors, and demonstrate the utility of the PC12 cell line for studies of tetanus toxicity and complex ganglioside expression.

Adrenal Gland Neoplasms↗
PubMedDate not supplied

Ganglioside changes associated with temporal lobe epilepsy in the human hippocampus.

To understand better the molecular and cellular events associated with status epilepticus, a multifaceted analysis has begun on hippocampal tissues therapeutically removed from patients with temporal lobe epilepsy. In this first study, quantitative changes in major ganglioside species are reported, as well as the immunocytochemical localization on the ganglioside GD3 in epileptic human hippocampus. Although significant variations were found between patients, the pattern of change was consistent when compared to normal values obtained from an autopsied specimen and the literature. Total ganglioside content was reduced in epileptic hippocampi, which was attributable, in part, to pyramidal cell loss found in CA1 and CA3. In each case, the percentage of ganglioside GD3 was increased significantly, while ganglioside GD1a decreased. The former change is probably associated with reactive astrocytosis and the latter with loss of neuronal dendrites. Immunocytochemical localization revealed GD3 in the stratum radiatum and the subgranular layer of the dentate gyrus. In these areas, GD3 was present in punctate structures and astrocytes. These findings indicate that GD3 increases in selected areas of the sclerotic hippocampus and is presumably related to localized accumulation of reactive glial cells. Since gangliosides have a high affinity for calcium and localized increase in extracellular calcium could disrupt normal neuronal function, the localized increase in GD3 may not only denote reactive glial cells but may contribute directly to the altered, hyperexcitable condition of epilepsy.

Chromatography, Thin Layer↗
PubMed1986

Human influenza A virus hemagglutinin distinguishes sialyloligosaccharides in membrane-associated gangliosides as its receptor which mediates the adsorption and fusion processes of virus infection. Specificity for oligosaccharides and sialic acids and the sequence to which sialic acid is attached.

Human influenza A virus isolates bearing antigenically different H1 (A/PR/8/34), H2 (A/Japan/305/57), and H3 (A/Aichi/2/68, A/X-31) hemagglutinin serotypes caused extensive hemagglutination, low pH fusion, and hemolysis of asialoerythrocytes reconstituted with gangliosides. Sialylparaglobosides (IV3NeuAc-nLc4Cer, IV6NeuAc-nLc4Cer), I-active and i-active (VI3NeuAc-nLc6Cer) gangliosides, and GM3-NeuAc commonly exhibited significant specific receptor activity toward the viruses. A/PR/8/34 recognized IV3NeuAc-nLc4Cer containing the NeuAc alpha 2-3Gal sequence preferentially over IV6NeuAc-nLc4Cer containing NeuAc alpha 2-6Gal, whereas the other two recognized the NeuAc alpha 2-6Gal sequence preferentially over NeuAc alpha 2-3Gal. Responsiveness of erythrocytes labeled with gangliosides containing NeuGc to the viruses used was considerably lower than that of erythrocytes labeled with gangliosides containing NeuAc. The activities of GM1a, GM2, and GD1b bearing NeuAc on inner galactose of the ganglio series core were also very low. These results indicate that sialyloligosaccharides of IV3NeuAc-nLc4Cer, IV6NeuAc-nLc4Cer, I-active ganglioside, and VI3NeuAc-nLc6Cer in addition to GM3-NeuAc and GM1b-NeuAc (Suzuki, Y., Matsunaga, M., and Matsumoto, M. (1985), J. Biol. Chem. 260, 1362-1365; Suzuki, Y., Matsunaga, M., Nagao, Y., Taki, T., Hirabayashi, Y., and Matsumoto, M. (1985) Vaccine 3, 201-203) are functional receptor determinants toward hemagglutinin of human influenza A viruses, and the viruses differentiate microdomains of the gangliosides, such as the sialic acid species (NeuAc, NeuGc) and the sequence of sialic acid linkages (NeuAc alpha 2-3Gal, NeuAc alpha 2-6Gal).

Animals↗
PubMed1986

Monoclonal antibody 18B8, which detects synapse-associated antigens, binds to ganglioside GT3 (II3 (NeuAc)3LacCer).

By immunofluorescence, mouse monoclonal antibody 18B8 detects developmentally regulated antigens in chick neural retina. In older embryos and in adults these antigens are localized in discrete laminae within the inner and outer synaptic layers. The antibody binds to several gangliosides that undergo both qualitative and quantitative changes during neuronal development (Grunwald, G.B., Fredman, P., Magnani, J.L., Trisler, D., Ginsburg, V., and Nirenberg, M. (1985) Proc. Natl. Acad. Sci. U.S.A. 82, 4008-4012). The simplest of these gangliosides was isolated from lipid extracts of 10-day chick embryonic retinas by DEAE-Sepharose and silicic acid column chromatography. About 300 micrograms was obtained from 9.3 g (wet weight) of retina. The isolated ganglioside was identified as GT3 by enzymatic analysis and by a comparison of its properties with the authentic ganglioside. By immunostaining thin-layer chromatograms with antibody 18B8, GT3 was detected in gangliosides from human neural tissue including cerebellum, optic nerve, and spinal cord, but not in gangliosides from human liver, pancreas, small intestine, adrenals, thyroid, or erythrocytes. GT3 was also found in five of seven human melanoma cell lines.

Animals↗
PubMed1985

Identification of a novel ganglioside on erythrocytes with blood group Cad specificity.

The blood group Cad antigen is a carbohydrate structure well characterized on the sialoglycoproteins of the red cell membrane from some rare individuals (Blanchard, D., Cartron, J. P., Fournet, B., Montreuil, J., Van Halbeck, H., and Vliegenthart, J.F.G. (1983) J. Biol. Chem. 258, 7691-7695). However, protease treatment of whole cells did not destroy their antigenic activity which indicated that glycolipid might also be involved in the antigenic reaction. A crude ganglioside fraction was prepared from Cad cells and found to inhibit the hemagglutination reaction, whereas neutral glycolipids were inactive. Further analysis of the ganglioside extract from Cad erythrocytes by thin layer chromatography revealed an unusual profile characterized by a lower content of sialosylparagloboside and the presence of a novel ganglioside of slower mobility. Immunochemical studies demonstrate that this ganglioside binds Helix pomatia lectin and inhibits human anti-Sda antibody. In addition, a ganglioside with identical chromatographic mobility can be obtained by the enzymatic transfer of GalNAc from UDP-GalNAc to sialosylparagloboside using a microsomal preparation from human kidney. These results together with cell surface labeling experiments suggest that the major ganglioside of Cad erythrocytes might be derived from sialosylparagloboside by substitution with an additional N-acetylgalactosamine residue.

Animals↗
PubMed1966

Changes in sphingosine and fatty acid components of the gangliosides in developing rat and human brain.

Rat brain increases in weight after birth in three stages: (I) rapidly for the first 2 weeks, (II) at a lower rate from 2 to 5 weeks, and (III) at a still lower rate from 5 weeks to 5 months. During the succeeding period, designated IV, it maintains constant weight up to 1 year of age. Brain ganglioside content increased linearly during I and II, more slowly during III, and diminished during IV. The appearance of measurable amounts of brain sphingomyelin and cerebroside succeeded that of ganglioside. Ceramide with C18-sphingosine and C18 fatty acid was found in a large proportion of all three sphingolipids upon their first appearance in measurable quantity. C18 fatty acid in cerebroside rapidly declined to a negligible level, while in gangliosides and sphingomyelin it declined slowly but remained the major fatty acid component. Cerebrosides and sphingomyelin contained C18-sphingosine almost exclusively at all stages of rat brain growth. Gangliosides contained C18-sphingosine almost exclusively at birth, but subsequently accumulated C20-sphingosine until they had nearly equal quantities of each base type. Changes in human brain gangliosides resemble those in rat. In Tay-Sachs disease, gangliosides have C18-sphingosine predominantly, and a high content of C18 fatty acid.

Adult↗
PubMed1983

Ganglioside composition is regulated during differentiation in the neuroblastoma X glioma hybrid cell line NG108-15.

Cellular differentiation of the neuroblastoma X glioma hybrid cell line NG108-15 was measured and correlated with quantitative changes in the cells' ganglioside composition. The degree of differentiation was measured using an enzymatic marker, choline acetyltransferase (CAT), which is responsible for neurotransmitter synthesis in this cell line. Differentiation of these cells is commonly induced by agents such as dibutyryl cyclic adenosine 3':5'-monophosphate (Bt2cAMP). However, in our studies, we observed that these cells "self-differentiated," in the absence of chemical inducers, when the cells became dense in culture. The differentiation marker, CAT specific activity, rose from 150 to more than 400 pmol/min/mg of protein as cell density increased, attaining a level higher than that achieved by treatment with Bt2cAMP. Differentiation of sparse cultures could be induced by conditioned medium removed from dense cultures. This effect was not due to depletion of a serum component from the medium by the cells, since it was not mimicked by serum depletion or inhibited by addition of fresh serum to the conditioned medium. These data suggest that cell density-dependent differentiation was caused by release of a factor from the cells which induced differentiation in a concentration-dependent manner. Gangliosides, therefore, were purified from sparse control cultures, dense cultures, and cultures treated with the differentiating agents Bt2cAMP, prostaglandin E1 (plus theophylline), or butyric acid. Quantitative thin layer chromatography revealed that all of the cultures contained the four gangliosides GM3, GM2, GM1, and GD1a. The concentration of one of the gangliosides, GM2, increased markedly (up to 12-fold) during differentiation. The GM2 concentration correlated closely with the level of CAT activity in the different cultures (r = 0.99). These data demonstrate that the ganglioside concentration in these cells is regulated during differentiation, a finding consistent with a possible role for gangliosides in the differentiated phenotype.

Alprostadil↗
PubMedDate not supplied

[Role of gangliosides in the processes of calcium ion transport across the membranes of nerve cells].

It has been demonstrated that the content of extracellular Ca in the nervous system is inversely related to the content of gangliosides. The results obtained on invertebrates, lower and higher vertebrates indicate that the highest content of extracellular Ca is typical of the nervous tissue of invertebrates, whereas the lower one--of the nervous tissue of higher vertebrates (mammals). Ganglioside content, on the contrary, is the highest in the brain tissue of the higher vertebrates (mammals and birds), being significantly lower in lower vertebrates; no gangliosides was found at all in the nervous tissue of protostomian invertebrates. The highest ganglioside content in the organism of vertebrates is characteristic to the surface membranes of the nervous cells, especially in the region of synapses. Functional significance of the inverse relationship between the content of extracellular Ca and gangliosides is discussed from the standpoint of one of the authors (R. Veh) who postulated the existence of calcium--ganglioside buffer in the vicinity of the surface of the nervous cells.

Amphibians↗
PubMed1984

Inhibition of mitogen- and antigen-induced lymphocyte activation by human leukemia cell gangliosides.

Human leukemia cell gangliosides were found to inhibit in vitro activation of human lymphocytes by lectins and the soluble antigen Candida albicans at concentrations as low as 2 micrograms/2 X 10(5) lymphocytes/0.2-ml cultures. The inhibition was not due to a reduction in the number of viable cells, but to an inhibition of blast formation. Three gangliosides were purified from the original mixture and tested for their ability to inhibit in vitro activation of lymphocytes. All 3 gangliosides inhibited lymphocyte activation to a similar degree when tested over a concentration range of 5 to 100 micrograms per ml of culture. Removal of sialic acid from the gangliosides by neuraminidase treatment significantly reduced or abolished their inhibitory effect. The gangliosides used in our analyses are not unique to leukemia cells. However, their concentration is increased in patients with cancers. Therefore, our results demonstrate that gangliosides isolated from human leukemia cells can inhibit lymphocyte activation when added in concentrations equivalent to those found in sera of patients with leukemia and other cancers.

Candida albicans↗
PubMed1983

[Dependence of ganglioside composition of leukemic lymphocytes on density cell population density].

A comparative study of gangliosides from bovine normal lymph lymphocytes and leukemic blood lymphocytes and from calf thymocytes incubated at different cell densities was carried out. The ganglioside composition of thymocytes and normal lymphocytes was insensitive to the variations in cell density. In the case of leukemic lymphocytes the decrease of cell density resulted in a reduction of the absolute and relative content of one of the major ganglioside components, disialosyllactosylceramide, and in a concomitant increase of its precursor, N-glycoloylneuraminosyllactosylceramide. The content of the third major ganglioside component, sialosylparagloboside, did not depend on cell density. The cell density dependent changes in the ganglioside composition of leukemic lymphocytes are qualitatively coincident with those observed at variable concentrations of blood lymphocytes in leukemic animals. A possible mechanism underlying the differences in ganglioside metabolism during cell density changes in leukemic lymphocytes is discussed.

Animals↗
PubMed1983

Gangliosides of hog skeletal muscle.

The ganglioside content of hog skeletal muscle was 27 nmol/g wet weight when calculated as lipid-bound sialic acid. The elution profile on DEAE-Sephadex A-25 column chromatography showed that the monosialoganglioside fraction (61% of total lipid-bound sialic acid) contained at least seven major gangliosides and that the disialoganglioside fraction (25%) contained four major ones. These gangliosides were purified by latrobeads column chromatography, and from the results of sugar analysis, specific enzymatic hydrolysis, a permethylation study and CrO3 oxidation, the structures of these gangliosides were determined to be as follows: A, GM3(NeuAc); B, GM3(NeuGc); C, sialosylparagloboside(NeuAc); D sialosylparagloboside (NeuGc); E, GM1(NeuAc); F, GM1(NeuGc); G, sialosyllactosaminylparagloboside(NeuAc) (monosialogangliosides) and H, GD1a(NeuAc, NeuAc); I, GD1a(NeuAc, NeuGc); J, GD1a(NeuGc, NeuGc); K, GD1b(NeuAc, NeuAc) (disialogangliosides), where NeuAc is N-acetylneuraminic acid and NeuGc is N-glycolylneuraminic acid as sialyl groups. The major fatty acids of gangliosides A, D and G were mainly C16:0, but those of the others, E, F, H, J and K, were C16:0, C18:0, C22:0 and C24:0. The long-chain bases were predominantly C18:1 sphingosine in all gangliosides.

Animals↗
PubMed1982

[Gangliosides of neural and extraneural tissues of various species of mammals].

The ganglioside patterns of the forebrain, cerebellum and brain stem from horse, donkey, mule and goat have been determined by thin-layer chromatography. GM1, GD1a, GD1b and GT1b are the four major brain gangliosides. N-acetylneuraminic acid as the predominant sialic acid (congruent to 97%) and traces of N-glycolyneuraminic acid were found. The four above mentioned major gangliosides were also found in the forebrain, cerebellum and brain stem of adult rats. This pattern is not modified in rats under stress situation (at 4 degrees C for 3 months). In other extraneural organs from rats such as liver and kidney the patterns differ from those of the neural organs. Pig spleen lymphocyte gangliosides were studied by thin-layer chromatography. Their patterns show remarkable differences in comparison to those of neural organs. These gangliosides were also separated by DEAE Sephadex chromatography into two peaks, I and II. Peak I contains mainly monosialogangliosides and peak II contains disialogangliosides. No trisialogangliosides were practically detected in the material. Sheep platelets contain GM3 as predominant ganglioside.

Animals↗