The isolation and partial characterization of glycolipids of normal human leucocytes

Miras, C. J.; Mantzos, J.; Levis, Gabriel M.

doi:10.1042/bj0980782

Cited by 62 publications

(13 citation statements)

References 23 publications

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“…The presence of a high proportion of 2-hydroxy fatty acids indicated a major difference from the fatty acid compositions of human leucocytes glycosphingolipids (1). The fatty acid compositions of the total glycosphingolipids of blood lymphocytes, thymus and lymph nodes, and thymus ceramides from one small male pig are shown in Table III.…”

Section: Resultsmentioning

confidence: 97%

“…Even so, there are wide variations in the amounts of these minor components in different tissues, and, compared with most tissues, the white blood cells are relatively rich in glycosphingolipids. The white cells, which have been examined in a limited number of species (1)(2)(3), contained predomi- nantly either dihexosyl or trihexosyl ceramides (GL-2, GL-3). It has been shown that glycolipid biosynthesis occurs in bone marrow cells (2), as well as in normal human leucocytes (4,5), and that in immature leucemic cells it occurs at an accelerated rate (5).…”

Section: Introductionmentioning

confidence: 99%

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Glycosphingolipids of white cells lymphoid tissue and bone marrow

Levis¹,

Kesse‐Elias²

1974

Lipids

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Glycosphingolipids of lymphoid tissues, bone marrow cells, mixed blood leucocytes, separated lymphocytes, and granulocytes from large white pigs were analyzed by thin layer and gas liquid chromatographies. The composition of the glycosphingolipids of thymus and blood leucocytes (mixed population) was similar, and trihexosyl ceramide (galactosyl‐[1–4]‐galactosyl‐[1–4]‐glucosyl‐[1–1]‐ceramide) was the major glycosphingolipid component of both tissues. The fatty acid fractions of all glycolipids from the two tissues were analyzed, and gross differences are discussed. Blood lymphocytes had a higher content of glycosphingolipids/mole phospholipid or mg protein than thymus lymphocytes obtained by a gentle washing of sliced tissue. Similar and more pronounced differences were obtained when the glycosphingolipid content of bone marrow cells (>50% polymorphonuclear neutrophils) was compared with that of blood polymorphonuclear neutrophils. In general, most of the blood leucocytes were richer in glycosphingolipids than most of the cells of the lymphoid tissues and bone marrow. These results indicate a marked difference in lipid composition between blood polymorphonuclear neutrophils and bone marrow cells. It is possible that certain of the biophysical properties which characterize blood polymorphonuclear neutrophils and which derive from changes in the cell periphery of immature granulocytes are connected with these differences in membrane lipid composition.

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Section: Resultsmentioning

confidence: 97%

Section: Introductionmentioning

confidence: 99%

Glycosphingolipids of white cells lymphoid tissue and bone marrow

Levis¹,

Kesse‐Elias²

1974

Lipids

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“…The acidic fractions of heart GSL were carefully examined for the presence of su!fatides, and it was concluded either that they are not present or that they constitute a very minor component. This is not surprising since sulfatides have not been detected in a variety of extraneural human tissues including skeletal muscle (2,13,(20)(21)(22). They are major components of the human kidney (24) and the thyroid (7).…”

Section: Discussionmentioning

confidence: 99%

“…The results of GSL were expressed in nmoles cal-Culated from the area of the glucose peaks (6,7). Sphingosine was determined (12) as described previously (13). The sialic acid content of the gangliosides was determined by the modified (14) method of Svennerholm (15).…”

Section: Analytical Proceduresmentioning

confidence: 99%

Isolation and partial characterization of the neutral glycosphingolipids and gangliosides of the human heart

Levis¹,

Karli²,

Moulopoulos³

1979

Lipids

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The glycosphingolipids (GSL) of the human heart muscle have been isolated from total lipids by column and thin layer chromatography and their sugars and fatty acids analyzed by gas liquid chromatography. Hearts from traffic victims were obtained at autopsy between 12 and 16 hr after death and dissected into parts (left and right ventricular walls, intraventricular septum and papillary muscle). The neutral GSL content for those parts of the hearts of two males aged 22 and one female aged 14 ranged from about 90 to 160 nmoles/g wet weight. Trihexosyl ceramide and globoside were the most abundant neutral GSL. Total ganglioside content was about 50 nmoles/g wet weight, and the most abundant gangliosides were partially characterized as GM3 and GM1; other mono-, di- and trisialogangliosides were also present. Differences in the content and composition of neutral GSL and gangliosides between the heart and other human tissues are discussed. It is concluded that the patterns of these two GSL fractions of the heart are more complex than those of the extraneural human tissues.

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“…The acidic lipid fraction was dried by evaporation of solvents under reduced pressure, followed by lyophilization, and the dry residue was submitted to mild alkaline treatment [ 5 ] , followed by Folch partition [26].…”

Section: Isolation and Purificationmentioning

confidence: 99%

Studies on Glycosphingolipid Biosynthesis by Lectin-Stimulated Human Lymphocytes

et al. 1980

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1. Human peripheral lymphocytes, normal and stimulated by phytohemagglutinin or concanavalin A, were investigated with respect to their ability to biosynthesize neutral and acidic glycosphingolipids from D‐[U‐14C]glucose, D‐[U‐14C]galactose and D‐[U14C]glucosamine as precursors. 2. Galactose and glucosamie are taken up selectively, in the presence of excessive glucose concentrations. Labeling of total neutral glycosphingolipids from D‐[U‐14C]galactose in normal cells decreases after the first 6 h while in stimulated cells there is a fourfold and a sevenfold increase after 6 and 12 h of incubation respectively. Under similar conditions stimulation with concanavalin A gives a fourfold increase after 12 h of incubation. 3. Analysis of individual glycosphingolipids biosynthesized from [U‐14C]galactose shows that lactosylceramide is the major radioactive neutral glycosphingolipid and that stimulation by phytohemagglutinin yields an almost sevenfold and 13‐fold increase in the radioactivity incorporated within 6 and 12 h of incubation respectively. Glucosylceramide shows about an eightfold increase, globotetraosylceramide a threefold increase and globotriaosylceramide a fourfold increase. The rate of incorporation into glucosylceramide of stimulated lymphocytes declines after 6 h of incubation, accompanied by a concomitant increase of incorporation into lactosylceramide. 4. At 12 h of incubation the ratios of radioactivities incorporated into neutral glycosphingolipids of phytohemagglutinin‐stimulated cells compared to normal cells were 1.0 for D‐[U‐14C]glucose 7.0 for D‐[U‐14C]galactose and 2.5 for D‐[U‐14C]glucosamine. Respective ratios for lactosylceramide are 1.0 for [U‐14C]glucose and 13.0 for [U‐14C]galactose. These differences did not arise from changes of the uptake of the glycosyl precursor by the cell due to stimulation. 5. Incorporation of D‐[U‐14C]galactose into II3‐N‐acetylneuraminosyllactosylceramide by cells stimulated by phytohemagglutinin and concanavalin A is enhanced 14‐times and 15‐times respectively. With [U‐14C]glucosamine as precursor, this increase in the labeling is much more impressive, 80‐fold after 12 h of incubation by the phytohemagglutinin‐stimulated lymphocytes. Neuraminidase treatment and gas radiochromatographic analysis of the labeled compound derived from [U‐14C]‐galactose as precursor indicated that 89% of the radioactivity was incorporated into the glucosyl and galactosyl moieties, in a ratio 1:1. With [U‐14C]glucosamine as precursor, a selective labeling of the sialyl moiety of the II3‐N‐acetylneuraminosyllactosylceramide was indicated. 6. The pattern of complex gangliosides (more complex than II3‐N‐acetylneuraminosyllactosylceramide) which are biosynthesized after phytohemagglutinin stimulation of the cells, show no significant differences when compared to the patterns obtained from normal cells.

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The isolation and partial characterization of glycolipids of normal human leucocytes

Cited by 62 publications

References 23 publications

Glycosphingolipids of white cells lymphoid tissue and bone marrow

Glycosphingolipids of white cells lymphoid tissue and bone marrow

Isolation and partial characterization of the neutral glycosphingolipids and gangliosides of the human heart

Studies on Glycosphingolipid Biosynthesis by Lectin-Stimulated Human Lymphocytes

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