The Distribution of Fe59 Tagged Human Erythrocytes in Centrifuged Specimens as a Function of Cell Age

Borun, E.Raymond; Figueroa, William G.; Perry, Seymour

doi:10.1172/jci103468

Cited by 186 publications

(75 citation statements)

References 10 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Antigen sensitive cells 10 hr after stimulation are much lighter than prior to antigenic stimulation (31) but plasma cells are, on the average, denser than either 19S PFC or total spleen cells (30). Erythrocytes become progressively more dense as they age (32).…”

Section: Discussionmentioning

confidence: 99%

Immunological Memory in Mice

L’age-Stehr

Herzenberg

1970

The Journal of Experimental Medicine

View full text Add to dashboard Cite

Plaque forming cells (PFC) of different immunoglobulin classes producing antibodies against sheep erythrocytes were separated according to their buoyant densities by means of equilibrium centrifugation in a stepwise BSA gradient. In the period of 7–10 days after immunization γM PFC are markedly enriched in fractions of low density and relatively depleted in fractions of high density. The distribution of total γG PFC shows less enrichment in the lower density fractions and less depletion in the higher density fractions. The density profile for γG2a PFC is even flatter, with a significant difference (depletion) relative to the unseparated spleen cells only in the highest density fraction. The density gradient distributions of cells able to transfer an adoptive immune response of the various immunoglobulin classes are markedly different from the PFC distribution. Cells obtained 7–10 days after immunization able to transfer an IgM response are present in the same proportions across the density gradient, whereas memory cells for γG2a obtained at this time are markedly enriched in fractions of low density and virtually depleted from high density fractions. With increasing time after primary immunization, the γG2a memory cells increase progressively in density and by 6 weeks the higher and lower density fractions have the same proportions of γG2a memory cells. The total γG (mainly γG1) memory cells by 7–10 days show slight enrichment in low density fractions and no depletion in high density fractions. The conclusions were reached that (a) memory for γG1 develops earlier than memory for γG2a and (b) that memory for anti-SRBC antibodies of different classes is carried in separate cells. When gradient fractions enriched for PFC and memory cells for all classes were completely depleted of PFC using glass bead columns, the ability of this fraction to transfer memory for all classes was not diminished. This shows that memory cells are not identical with cells secreting antibodies.

show abstract

Section: Discussionmentioning

confidence: 99%

Immunological Memory in Mice

L’age-Stehr

Herzenberg

1970

The Journal of Experimental Medicine

View full text Add to dashboard Cite

show abstract

“…Erythrocytes of subjects with the G6PD-.B isoenzyme were separated into young and old cells by centrifugation [3] and washed in 0.01 M Tris HC1 buffer, pH 7.4, that contained 2.5 mM MgCl 2 , 0.14 M KC1, 0.14 mM 2-mercaptoethanol, and either 2.5, 10, or 15 fjM NADP. Cells were suspended in 9 volumes of the buffer in which they were washed, and they were lysed by freezing and thawing.…”

Section: Methodsmentioning

confidence: 99%

Change in Electrophoretic Mobility of Glucose-6-Phosphate Dehydrogenase with Aging of Erythrocytes

1972

View full text Add to dashboard Cite

ExtractElectrophoretic properties of glucose-6-phosphate dehydrogenase (G6PD) were studied in mother-infant pairs and in young and old erythrocytes of adults. G6PD isoenzymes of erythrocytes of newborn infants migrated at rates 3-4% faster than the corresponding isoenzymes of maternal erythrocytes. Analysis of hemolysates of young and old erythrocytes of adults revealed that G6PD of young cells also moved about 3% faster than that of old cells. In contrast, active enzyme sedimentation analysis revealed that the G6PD enzyme proteins of old and young erythrocytes of adults, of newborns, and of their mothers sedimented with essentially identical rates (S 2 o,w = 6.8). Migration in gels of different acrylamide concentrations and velocity sedimentation analysis indicated that the G6PD of old and of young erythrocytes was in the same state of aggregation. Extensive equilibration of the hemolysates with nicotinamide-adenine dinucleotide phosphate (NADP) by filtration through Sephadex G-25 induced a clear increase in the migration rate of the G6PD of old cells of adults and of mothers at term to the rate characteristic of the enzyme of neonates and of young cells of adults. Experiments in which NADP was labeled with 14 G indicated that, in the process of gel nitration, the NADP content of the aged G6PD was restored. Thus the rate of migration of G6PD in acrylamide gel seems to be a function of the amount of NADP bound.

show abstract

“…This provides a way to see what the relationship is between oharbumet~,Na and MCV within the cell population distribution of the different individual bloods, and whether a subpopulation of cells with very high or low values of o~b,met,,,Na exists. The fractionation of red cells according to density is based on changes in the MCHC at almost constant MCH (Borun et al, 1957;Hoffman, 1958;Murphy et al, 1974;Lee et al, 1984). The increase in MCHC with cell density presumably reflects a loss of solutes (e.g., Ki) and cell water, causing a decrease in Ouabain-sensitive Na/K fluxes were measured at V,..x in cells after modification of Nai and I~ with the nystatin technique as described in Methods.…”

Section: Characteristics Of Fractionated Red Cellsmentioning

confidence: 99%

Internal magnesium, 2,3-diphosphoglycerate, and the regulation of the steady-state volume of human red blood cells by the Na/K/2Cl cotransport system.

Mairbäurl

Hoffman

1992

The Journal of General Physiology

View full text Add to dashboard Cite

This study is concerned with the relationship between the Na/K/CI cotransport system and the steady-state volume (MCV) of red blood cells. Cotransport rate was determined in unfractionated and density-separated red cells of different MCV from different donors to see whether cotransport differences contribute to the difference in the distribution of MCVs. Cotransport, studied in cells at their original MCVs, was determined as the bumetanide (10 ~.M)-sensitive 2~Na efflux in the presence of ouabain (50 I~M) after adjusting cellular Na (Nai) and K~ to achieve near maximal transport rates. This condition was chosen to rule out MCV-related differences in Nai and Ki that might contribute to differences in the net chemical driving force for cotransport. We found that in both unfractionated and density-separated red cells the cotransport rate was inversely correlated with MCV. MCV was correlated directly with red cell 2,3-diphosphoglycerate (DPG), whereas total red cell Mg was only slightly elevated in cells with high MCV. Thus intraceUular free Mg (Mg~ e) is evidently lower in red cells with high 2,3-DPG (i.e., high MCV) and vice versa. Results from flux measurements at their original MCVs, after altering Mg[ r~ with the ionophore A23187, indicated a high Mgi sensitivity of cotransport: depletion of M~ r~ inhibited and an elevation of M~ tee increased the cotransport rate. The apparent K05 for Mg~ e" was ~ 0.4 mM. Maximizing Mig~ ~ at optimum Nai and I~ minimized the differences in cotransport rates among the different donors. It is concluded that the relative cotransport rate is regulated for cells in the steady state at their original cell volume, not by the number of copies of the cotransporter but by differences in Mig~ ~. The interindividual differences in Mg~ ~, determined primarily by differences in the 2,3-DPG content, are responsible for the differences in the relative cotransport activity that results in an inverse Address reprint requests to Dr.

show abstract

The Distribution of Fe59 Tagged Human Erythrocytes in Centrifuged Specimens as a Function of Cell Age

Cited by 186 publications

References 10 publications

Immunological Memory in Mice

Immunological Memory in Mice

Change in Electrophoretic Mobility of Glucose-6-Phosphate Dehydrogenase with Aging of Erythrocytes

Internal magnesium, 2,3-diphosphoglycerate, and the regulation of the steady-state volume of human red blood cells by the Na/K/2Cl cotransport system.

Contact Info

Product

Resources

About