Permeation of the luminal capillary glycocalyx is determined by hyaluronan

1999 ◽  
Vol 277 (2) ◽  
pp. H508-H514 ◽  
Author(s):  
Charmaine B. S. Henry ◽  
Brian R. Duling
Keyword(s):  
Blood Cell ◽  
Red Blood Cells ◽  
Red Blood Cell ◽  
Blood Cells ◽  
Enzyme Treatment ◽  
Apical Surface ◽  
Bright Field ◽  
Endothelial Surface ◽  
The Difference

The endothelial cell glycocalyx influences blood flow and presents a selective barrier to movement of macromolecules from plasma to the endothelial surface. In the hamster cremaster microcirculation, FITC-labeled Dextran 70 and larger molecules are excluded from a region extending almost 0.5 μm from the endothelial surface into the lumen. Red blood cells under normal flow conditions are excluded from a region extending even farther into the lumen. Examination of cultured endothelial cells has shown that the glycocalyx contains hyaluronan, a glycosaminoglycan which is known to create matrices with molecular sieving properties. To test the hypothesis that hyaluronan might be involved in establishing the permeation properties of the apical surface glycocalyx in vivo, hamster microvessels in the cremaster muscle were visualized using video microscopy. After infusion of one of several FITC-dextrans (70, 145, 580, and 2,000 kDa) via a femoral cannula, microvessels were observed with bright-field and fluorescence microscopy to obtain estimates of the anatomic diameters and the widths of fluorescent dextran columns and of red blood cell columns (means ± SE). The widths of the red blood cell and dextran exclusion zones were calculated as one-half the difference between the bright-field anatomic diameter and the width of the red blood cell column or dextran column. After 1 h of treatment with active Streptomyces hyaluronidase, there was a significant increase in access of 70- and 145-kDa FITC-dextrans to the space bounded by the apical glycocalyx, but no increase in access of the red blood cells or in the anatomic diameter in capillaries, arterioles, and venules. Hyaluronidase had no effect on access of FITC-Dextrans 580 and 2,000. Infusion of a mixture of hyaluronan and chondroitin sulfate after enzyme treatment reconstituted the glycocalyx, although treatment with either molecule separately had no effect. These results suggest that cell surface hyaluronan plays a role in regulating or establishing permeation of the apical glycocalyx to macromolecules. This finding and our prior observations suggest that hyaluronan and other glycoconjugates are required for assembly of the matrix on the endothelial surface. We hypothesize that hyaluronidase creates a more open matrix, enabling smaller dextran molecules to penetrate deeper into the glycocalyx.

scholarly journals Accounting for red blood cell accessibility reveals distinct invasion strategies inPlasmodium falciparumstrains

2019 ◽  
Author(s):  
Francisco Cai ◽  
Tiffany M. DeSimone ◽  
Elsa Hansen ◽  
Cameron V. Jennings ◽  
Amy K. Bei ◽  
...  
Keyword(s):  
Blood Cell ◽  
Red Blood Cells ◽  
Red Blood Cell ◽  
Blood Cells ◽  
Cell Structure ◽  
Clinical Manifestations ◽  
Parasite Growth ◽  
Invasion Pathways ◽  
Invasion Rate

AbstractThe growth of the malaria parasitePlasmodium falciparumin human blood causes all clinical manifestations of malaria, a process that begins with the invasion of red blood cells. Parasites enter red blood cells using distinct pairs of parasite ligands and host receptors that define particular invasion pathways. Parasite strains have the capacity to switch between invasion pathways. This flexibility is thought to facilitate immune evasion against particular parasite ligands, but may also reflect the fact that red blood cell surfaces are dynamic and composed of heterogeneous invasion targets. Different host genetic backgrounds affecting red blood cell structure have long been recognized to impact parasite growthin vivo, but even within a host, red blood cells undergo dramatic changes in morphology and receptor density as they age. The consequences of these heterogeneities for parasite growthin vivoremain unclear. Here, we measured the ability of laboratory strains ofP. falciparumrelying on distinct invasion pathways to enter red blood cells of different ages. We estimated invasion efficiency while accounting for the fact that even if the red blood cells display the appropriate receptors, not all are physically accessible to invading parasites. This approach revealed a tradeoff made by parasites between the fraction of susceptible cells and their invasion rate into them. We were able to distinguish between “specialist” strains exhibiting high invasion rate in fewer cells versus “generalist” strains invading less efficiently into a larger fraction of cells. We developed a mathematical model to predict that infection with a generalist strain would lead to higher peak parasitemiasin vivowhen compared with a specialist strain with similar overall proliferation rate. Thus, the heterogeneous ecology of red blood cells may play a key role in determining the rate of parasite proliferation between different strains ofP. falciparum.

scholarly journals Biconcave shape of human red-blood-cell ghosts relies on density differences between the rim and dimple of the ghost's plasma membrane

2016 ◽  
Vol 113 (51) ◽  
pp. 14847-14851 ◽  
Author(s):  
Joseph F. Hoffman
Keyword(s):  
Blood Cell ◽  
Red Blood Cells ◽  
Red Blood Cell ◽  
Blood Cells ◽  
Theoretical Work ◽  
Centrifugal Field ◽  
Hypotonic Hemolysis ◽  
Osmotic Solution ◽  
Dense Band ◽  
The Difference

The shape of the human red blood cell is known to be a biconcave disk. It is evident from a variety of theoretical work that known physical properties of the membrane, such as its bending energy and elasticity, can explain the red-blood-cell biconcave shape as well as other shapes that red blood cells assume. But these analyses do not provide information on the underlying molecular causes. This paper describes experiments that attempt to identify some of the underlying determinates of cell shape. To this end, red-blood-cell ghosts were made by hypotonic hemolysis and then reconstituted such that they were smooth spheres in hypo-osmotic solutions and smooth biconcave discs in iso-osmotic solutions. The spherical ghosts were centrifuged onto a coated coverslip upon which they adhered. When the attached spheres were changed to biconcave discs by flushing with an iso-osmotic solution, the ghosts were observed to be mainly oriented in aflatalignment on the coverslip. This was interpreted to mean that, during centrifugation, the spherical ghosts were oriented by a dense band in its equatorial plane, parallel to the centrifugal field. This appears to be evidence that the difference in the densities between the rim and the dimple regions of red blood cells and their ghosts may be responsible for their biconcave shape.

scholarly journals The Effects of Concentrated Eluted Anti-Red Blood Cell Antibodies on the in Vivo Survival of Normal Red Blood Cells

Blood ◽  
1960 ◽  
Vol 15 (4) ◽  
pp. 525-533 ◽  
Author(s):  
NEIL W. CULP ◽  
HUGH CHAPLIN
Keyword(s):  
Blood Cell ◽  
Red Blood Cells ◽  
Hemolytic Anemia ◽  
Normal Control ◽  
Red Blood Cell ◽  
Blood Cells ◽  
Red Cells ◽  
Significant Alteration ◽  
Red Cell

Abstract 1. A method has been described for the preparation and sterilization of a concentrated eluate from human red cell stroma. 2. Red cells sensitized by such an eluate prepared from normal control red cells showed entirely normal in vivo survival, as did cells sensitized by eluate from anti-H coated cells. 3. Sensitization of red cells by concentrated eluates from a patient with Coombs-negative acquired hemolytic anemia and from a patient with Coombs-positive acquired hemolytic anemia did not cause significant alteration in the in vivo survival of the red cells. 4. Red cells sensitized by the concentrated eluate from anti-D sensitized cells disappeared from the recipient’s circulation very rapidly and were sequestered in the spleen, indicating preservation of the physiologic properties of the antibody throughout the elution, concentration and sterilization procedures.

scholarly journals Vesiculation of Red Blood Cells in the Blood Bank: A Multi-Omics Approach towards Identification of Causes and Consequences

Proteomes ◽  
2020 ◽  
Vol 8 (2) ◽  
pp. 6
Author(s):  
Joames K. Freitas Leal ◽  
Edwin Lasonder ◽  
Vikram Sharma ◽  
Jürgen Schiller ◽  
Giuseppina Fanelli ◽  
...  
Keyword(s):  
Blood Cell ◽  
Red Blood Cells ◽  
Red Blood Cell ◽  
Blood Cells ◽  
Blood Bank ◽  
Cell Aging ◽  
Lipid Organization ◽  
Membrane Components

Microvesicle generation is an integral part of the aging process of red blood cells in vivo and in vitro. Extensive vesiculation impairs function and survival of red blood cells after transfusion, and microvesicles contribute to transfusion reactions. The triggers and mechanisms of microvesicle generation are largely unknown. In this study, we combined morphological, immunochemical, proteomic, lipidomic, and metabolomic analyses to obtain an integrated understanding of the mechanisms underlying microvesicle generation during the storage of red blood cell concentrates. Our data indicate that changes in membrane organization, triggered by altered protein conformation, constitute the main mechanism of vesiculation, and precede changes in lipid organization. The resulting selective accumulation of membrane components in microvesicles is accompanied by the recruitment of plasma proteins involved in inflammation and coagulation. Our data may serve as a basis for further dissection of the fundamental mechanisms of red blood cell aging and vesiculation, for identifying the cause-effect relationship between blood bank storage and transfusion complications, and for assessing the role of microvesicles in pathologies affecting red blood cells.

scholarly journals CATECHOLAMINE-ACTIVATED SODIUM/PROTON EXCHANGE IN THE RED BLOOD CELLS OF THE MARINE TELEOST GADUS MORHUA

1994 ◽  
Vol 192 (1) ◽  
pp. 253-267 ◽  
Author(s):  
M Berenbrink ◽  
C Bridges
Keyword(s):  
Blood Cell ◽  
Red Blood Cells ◽  
Red Blood Cell ◽  
Blood Cells ◽  
Gadus Morhua ◽  
Atlantic Cod ◽  
Proton Exchange ◽  
Buffer System ◽  
Sodium Proton Exchanger

The effects of catecholamines on the pH and the cellular ion and water content were investigated in red blood cells from the Atlantic cod (Gadus morhua). Noradrenaline induced a rapid decrease in the extracellular pH (pHe) of red blood cells suspended in a CO2/bicarbonate or in a CO2/bicarbonate-free buffer system. The noradrenaline-induced changes in pHe were a saturable function of the external sodium ion concentration and were inhibited by amiloride but not by DIDS (4,4'-diisothiocyanatostilbene-2,2'-disulphonic acid, final concentration of both 10(-4) mol l-1). The catecholamine-induced extracellular acidification was accompanied by an intracellular alkalization and protons were moved from their electrochemical equilibrium. Proton extrusion was associated with an increase in the red blood cell sodium and chloride concentrations. In the presence of DIDS, the chloride movements were blocked and the net proton efflux under these conditions matched the net sodium influx. The results strongly suggested the activation of a sodium/proton exchanger by catecholamines in the red blood cells of the Atlantic cod. The red blood cell receptor affinity for adrenaline was three times higher than that for noradrenaline. Comparison with data in the literature for in vivo catecholamine concentrations indicated that adrenaline was more effective than noradrenaline in activating the red blood cell sodium/proton exchanger in the Atlantic cod in vivo.

scholarly journals Quantitation of immunoglobulin associated with senescent erythrocytes from the rabbit

Blood ◽  
1991 ◽  
Vol 77 (5) ◽  
pp. 1096-1099 ◽  
Author(s):  
GL Dale ◽  
RB Daniels
Keyword(s):  
Blood Cell ◽  
Red Blood Cells ◽  
Red Blood Cell ◽  
Blood Cells ◽  
Antibody Binding ◽  
Biochemical Processes ◽  
Senescent Erythrocytes

The biochemical processes that determine the lifespan of mammalian erythrocytes are unknown; one prominent theory suggests that antibody binding to the senescent red blood cell identifies it for removal from the circulation. To address this question, we have used a newly developed procedure for the isolation of aged erythrocytes that involves the biotinylation of rabbit red blood cells, in vivo aging of these cells, and the eventual in vitro recovery of the aged red blood cells by their affinity for an avidin support. Erythrocytes isolated with this method were found to have near-normal levels of cell- associated Ig throughout their 60-day lifespan. These data suggest that IgG accumulation is not part of the normal senescence process for erythrocytes in rabbits.

scholarly journals Validation of Daraex to Resolve Daratumumab-Induced Interferences in Pre-Transfusion Screen Tests

Blood ◽  
2019 ◽  
Vol 134 (Supplement_1) ◽  
pp. 4983-4983
Author(s):  
Maria Tenorio ◽  
Gemma Moreno Jiménez ◽  
Valentín García Gutiérrez ◽  
Ana Jiménez ◽  
Maria Jesús Blanchard ◽  
...  
Keyword(s):  
Blood Cell ◽  
Red Blood Cells ◽  
Red Blood Cell ◽  
Blood Cells ◽  
Research Funding ◽  
Screen Test ◽  
New Method ◽  
Negative Controls

Daratumumab is a CD38-directed antibody increasingly used for the treatment of adult patients with multiple mieloma. The membrane of red blood cells express CD38 and thus samples from patients treated with daratumumab show agglutination in red blood cell antibody screen tests performed prior to transfusion. This interference hinders the detection of red blood cell alloantibodies. Published literature has described a method to eliminate CD38 in red blood cells with DTT (Chapuy, 2016). This technique is cumbersome, requires positive and negative controls as DTT destroys Kell antigens and can produce in vitro hemolysis. The increasing number of multiple myeloma patients treated with daratumumab poses the need for a simple and straightforward technique with applicability in standard transfusion centers. DaraEx (Inno-Train) is a new anti-CD38 neutralizing agent that overcomes daratumumab-induced interferences detected in pre-transfusion tests without the major drawbacks associated with the DTT technique. Our aim was to validate and implement DaraEx as the method of choice to solve daratumumab interferences detected in pre-transfusion screen tests in a tertiary care center. A two-step approach using in vitro and in vivo samples was designed to validate the new method. First, we compared DaraEx efficacy in vitro to the reference DTT method in two samples spiked with daratumumab to achieve a concentration of 10mg/mL (Sample A: serum from a patient without known red blood cell alloantibodies; Sample B: serum from a patient with alloantibody anti-c). Red blood cells in the screen test (3 red blood cell screen; ID-DiaCell I-II-III) as well as positive (E+ red blood cells) and negative controls (K+ red blood cells) were treated with DTT 0.2M solution for 30 minutes at 37ºC and then washed four times with saline. In parallel, red blood cells in the screen test were incubated during 30 minutes at room temperature in a shaker (600rpm) with DaraEx. Red blood cells treated with each of these methods were used for indirect antiglobulin test with our gel card system (BioRad; IH-1000). Preference of method in terms of time needed and result interpretation was evaluated by three hematologists specialized in blood banking and four different technicians. Secondly, we tested pre-transfusion samples from patients treated with daratumumab with the DaraEx technique to check in vivo efficacy. There was a 100% concordance between both techniques (DDT reference method and DaraEx new method) in both in vitro samples. All hematologists and technicians found the DaraEx technique less cumbersome in terms of processing and time to result (2 hours with DTT versus 1 hour with DaraEx) and the interpretation straightforward. Twelve samples with daratumumab-induced interference in pre-transfusion screen tests belonging to 5 patients were tested between January and July 2019. All the interferences detected resolved with DaraEx regardless of time from last daratumumab administration (range: 7-145 days; mean: 57 days). Figure 1 shows screen test with and without treatment with DaraEx in a patient sample. In our experience, DaraEx technique is a simple, fast and efficacious method, regardless of time from last daratumumab administration, to resolve interferences secondary to daratumumab administration without the major disadvantages associated with DTT. Figure 1 Disclosures García Gutiérrez: Pfizer: Honoraria, Research Funding; Incyte: Honoraria, Research Funding; Novartis: Honoraria, Research Funding; BMS: Honoraria, Research Funding.

scholarly journals Quantitation of immunoglobulin associated with senescent erythrocytes from the rabbit

Blood ◽  
1991 ◽  
Vol 77 (5) ◽  
pp. 1096-1099 ◽  
Author(s):  
GL Dale ◽  
RB Daniels
Keyword(s):  
Blood Cell ◽  
Red Blood Cells ◽  
Red Blood Cell ◽  
Blood Cells ◽  
Antibody Binding ◽  
Biochemical Processes ◽  
Senescent Erythrocytes

Abstract The biochemical processes that determine the lifespan of mammalian erythrocytes are unknown; one prominent theory suggests that antibody binding to the senescent red blood cell identifies it for removal from the circulation. To address this question, we have used a newly developed procedure for the isolation of aged erythrocytes that involves the biotinylation of rabbit red blood cells, in vivo aging of these cells, and the eventual in vitro recovery of the aged red blood cells by their affinity for an avidin support. Erythrocytes isolated with this method were found to have near-normal levels of cell- associated Ig throughout their 60-day lifespan. These data suggest that IgG accumulation is not part of the normal senescence process for erythrocytes in rabbits.

scholarly journals The Entry of Sodium into Human Red Blood Cells in Vivo

1966 ◽  
Vol 50 (1) ◽  
pp. 75-88 ◽  
Author(s):  
L. J. Beilin ◽  
D. Eyeions ◽  
G. Hatcher ◽  
G. J. Knight ◽  
A. D. Munro-Faure ◽  
...  
Keyword(s):  
Blood Cell ◽  
Red Blood Cells ◽  
Red Blood Cell ◽  
Blood Cells ◽  
Compartmental Analysis ◽  
Analogue Computer ◽  
Packed Red Blood Cells ◽  
Human Red Blood Cells ◽  
In Series

The kinetics of sodium, movement into human red blood cells has been studied in vivo with 24Na. When human serum albumin-131I is used to measure the percentage of plasma trapped in the packed red blood cells after centrifugation, approximately 30 % of red blood cell sodium is found to equilibrate immediately with plasma. It is concluded that this immediately exchangeable compartment of red blood cell sodium is an experimental artefact, associated with the use of labeled albumin for measuring plasma trapping. This immediately exchangeable fraction disappears when sucrose-14C is used to measure plasma trapping. The experimental results were examined by compartmental analysis, using an analogue computer. The results obtained, when plasma trapping was measured with sucrose-14C could be simulated by the use of models containing two compartments, arranged in series or in parallel. The errors of the techniques used and the possible physical basis for the results are discussed.

scholarly journals The Measurement of Sodium Concentration in Human Red Blood Cells

1966 ◽  
Vol 50 (1) ◽  
pp. 61-74 ◽  
Author(s):  
L. J. Beilin ◽  
G. J. Knight ◽  
A. D. Munro-Faure ◽  
J. Anderson
Keyword(s):  
Blood Cell ◽  
Red Blood Cells ◽  
Red Blood Cell ◽  
Blood Cells ◽  
Sodium Concentration ◽  
Plasma Sodium ◽  
Human Red Blood Cells ◽  
Relative Centrifugal Force ◽  
The Difference ◽  
Extracellular Sodium

Experiments are described which indicate that iodinated human serum albumin underestimates the amount of extracellular sodium trapped in the packed layer of red blood cells, when cells and plasma are separated by centrifugation. Sucrose-14C also underestimates the amount of trapped extracellular sodium, but the difference between the percentages of sucrose-14C and extracellular sodium trapped is constant and independent of mean relative centrifugal force. It is concluded that human red blood cell sodium concentration can be measured with accuracy (a) if trapped plasma sodium is estimated with radioisotopes of sodium and a correction made for entry of sodium into the cells, providing cells and plasma can be separated rapidly; (b) by the use of sucrose as a standard plasma marker to derive the amount of trapped plasma sodium; (c) by washing the cells with sodium-free solutions. Reported values for red blood cell sodium concentration in healthy adults are critically reviewed.

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