Reduction of Prion Infectivity in Packed Red Cells by Separation of Whole Blood and Washing of the Red Cell Fraction.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 2890-2890
Author(s):  
Rodrigo Morales ◽  
Kimberley A. Buytaert-Hoefen ◽  
Eric T. Hansen ◽  
Dennis Hlavinka ◽  
Raymond Goodrich ◽  
...  
Keyword(s):  
Blood Cell ◽  
Red Blood Cell ◽  
Whole Blood ◽  
Red Cells ◽  
Cell Fraction ◽  
Health Concern ◽  
Red Cell ◽  
New Variant

Abstract Although prion diseases are rare in humans, the established link between a new variant form of CJD (vCJD) and the consumption of cattle meat contaminated by BSE have raised concerns about a possible outbreak of a large epidemic in the human population. Over the past few years, BSE has become a significant health concern in several countries, and it now seems apparent that vCJD can also be iatrogenically transmitted from human to human by blood transfusion. Exacerbating this state of affairs is the lack of a reliable test to identify individuals incubating the disease during the long and silent period from the onset of infection to the appearance of clinical symptoms. The purpose of this research study was to evaluate the effectiveness of separation of whole blood and washing of the red cell fraction for the removal of infectious scrapie prion protein (PrPSc) from red blood cell (RBC) suspensions. Samples of human, whole blood were spiked with 5 × 106 LD50 263K PrPSc. Analysis of the treated sample supernatants by Western blot revealed that approximately >88% of the PrPSc was removed with the initial plasma expression and the equivalent of 6% was detected in a saline wash (300 mL; 0.9% saline). The final sample of RBCs revealed no detectable levels of PrPSc by Western blots. Further analysis of the treated RBCs using the PMCA assay indicated detectable amounts of PrPSc only after 2 consecutive amplification rounds. Semi-quantitative analysis of PMCA amplification enabled us to estimate that the treated RBCs contained less than 1 × 104 LD50 PrPSc. This corresponded to removal levels exceeding ≥99% of spiked material in whole blood. These in vitro estimations were confirmed by in vivo infectivity studies in a hamster model of disease transmission. Results from in vivo studies displayed significant differences in the incubation periods of the spiked blood inoculated hamsters (100.1 ± 1.7) versus washed RBCs (135.8 ± 6.7). Moreover, a substantial difference in the attack rate (6/15: 40% in washed RBC, versus 13/13: 100% in spiked blood) further indicated a substantial removal of infectious prions. Comparison of this data with results of animals inoculated with different dilutions of infectious material, indicated a >99.94% reduction of infectivity. Washed, packed human red cells produced by this procedure were able to be stored in standard additive solutions (AS-3) for 42 days while still meeting all in vitro blood bank standards for acceptable red cell quality. Conclusion This data suggests that separation of plasma coupled with a simple, low volume wash of red cells may represent an efficient method to remove prions from red blood cell fractions, thus reducing possible infectivity of these products.

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 Regulatory T-cell status in red cell alloimmunized responder and nonresponder mice

Blood ◽  
2009 ◽  
Vol 113 (22) ◽  
pp. 5624-5627 ◽  
Author(s):  
Weili Bao ◽  
Jin Yu ◽  
Susanne Heck ◽  
Karina Yazdanbakhsh
Keyword(s):  
T Cells ◽  
Blood Cell ◽  
Red Blood Cell ◽  
Major Complication ◽  
Glycophorin A ◽  
Red Cell ◽  
Suppressive Activity ◽  
Human Glycophorin

Abstract Red blood cell alloimmunization remains a major complication for transfusion-dependent patients, but immune factors governing risk for alloimmunization are unknown. We hypothesized that CD4+ regulatory T cells (Tregs), which we have shown control the rate and the frequency of red blood cell alloimmunization in mouse models, may dictate responder/nonresponder status. Using a transfusion regimen in which more than 50% of mice develop alloantibodies to human glycophorin A antigen, we found reduced in vitro and in vivo Treg-suppressive activity in responders compared with nonresponders that was the result of impaired Treg suppressor function. Moreover, responders were prone to developing additional alloantibodies to strong immunogens, whereas nonresponders were resistant to alloimmunization. Altogether, our data raise the possibility that Treg activity may be used as a marker for identifying responder/nonresponder status in transfusion recipients.

scholarly journals Resistance of Reticulocytes and Young Erythrocytes to the Action of Specific Hemolytic Serum

Blood ◽  
1952 ◽  
Vol 7 (6) ◽  
pp. 602-606 ◽  
Author(s):  
W. O. CRUZ ◽  
P. C. JUNQUEIRA
Keyword(s):  
Blood Cell ◽  
Red Blood Cell ◽  
Normal Animal ◽  
Red Cells ◽  
Normal Adult ◽  
Severe Anemia ◽  
Red Cell ◽  
Photoelectric Method ◽  
The Relationship

Abstract A photoelectric method is used to determine the degree of hemolysis produced by specific hemolytic serum on erythrocytes of dogs, taking as a standard the amount required for 50 per cent hemolysis. The requirement of the normal dog red blood cell is 1.2 cu. ml. (±0.7 cu. ml.) of hemolytic serum, under the conditions described in the method. The red cells of dogs given a sufficient dose of acetylphenylhydrazine to produce a severe anemia, followed by an intense period of blood regeneration, required an amount of hemolytic serum about two to four times greater than in the normal animal (values from 3.0 to 4.8 cu. ml. of hemolytic serum are recorded). This result shows that the reticulocytes and young erythrocytes are much more resistant than the normal adult red cell to the action of hemolytic serum. It was possible by this technic to concentrate the reticulocytes in vitro, destroying selectively the adult erythrocytes of a sample of blood by an appropriate amount of specific hemolytic serum. The relationship between these results and the high reticulocytosis observed during crisis in hemolytic jaundice is discussed.

Characterization of red blood cell metabolism in rainbow trout

1990 ◽  
Vol 154 (1) ◽  
pp. 475-489 ◽  
Author(s):  
P. J. Walsh ◽  
C. M. Wood ◽  
S. Thomas ◽  
S. F. Perry
Keyword(s):  
Rainbow Trout ◽  
Blood Cell ◽  
Red Blood Cell ◽  
Whole Blood ◽  
Cell Metabolism ◽  
Plasma Concentrations ◽  
Co2 Production ◽  
Production Rates

Red blood cell metabolism was studied in vitro using whole blood obtained by catheter from resting rainbow trout (Oncorhynchus mykiss). Preparations were viable as shown by stable NTP, metabolite and catecholamine levels and acid-base status, all of which remained at in vivo levels over the 2 h incubation period. Enzymes diagnostic of glycolysis, the tricarboxylic acid (TCA) cycle and phosphagen metabolism were all present in significant amounts in red blood cells. In direct comparisons of 14C-labelled substrates at normal resting plasma concentrations, rates of CO2 production were in the order: glucose greater than lactate greater than alanine greater than oleate. Total CO2 production rates from these four oxidative substrates did not equal directly measured O2 consumption rates, indicating that other substrates may also be important in vivo. Oxidative pathway Km values for glucose (8.4 mmol l-1), lactate (3.3 mmol l-1) and alanine (0.8 mmol l-1) were well within the normal physiological ranges of plasma concentrations. Glucose concentration did not affect lactate oxidation rates, but there was some inhibition (27%) of glucose oxidation by high lactate concentrations (20 mmol l-1). The observed Km values and competitive interactions suggest that changes in plasma concentrations associated with environmental stresses can considerably alter the relative rates of oxidation of glucose and lactate in vivo. Considerable pentose-phosphate shunt activity was detected in red cells, as indicated by high activities of glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase and high CO2 production rates from (1–14C)-labelled glucose. Even in the presence of normal O2 levels, a significant percentage (28%) of glucose metabolism was directed to lactate production. Taken together, these results demonstrate that rainbow trout whole blood incubated in vitro constitutes a dynamic and viable system for metabolic studies at the pathway level.

LACK OF EFFECT OF CORTICOSTEROIDS ON THE IN VIVO DISAPPEARANCE OF SULFHYDRYL-INHIBITED AND ANTIBODY-COATED ERYTHROCYTES

1964 ◽  
Vol 47 (3_Suppl) ◽  
pp. S28-S36
Author(s):  
Kailash N. Agarwal
Keyword(s):  
Red Cells ◽  
List Type ◽  
Red Cell

ABSTRACT Red cells were incubated in vitro with sulfhydryl inhibitors and Rhantibody with and without prior incubation with prednisolone-hemisuccinate. These erythrocytes were labelled with Cr51 and P32 and their disappearance in vivo after autotransfusion was measured. Prior incubation with prednisolone-hemisuccinate had no effect on the rate of red cell disappearance. The disappearance of the cells was shown to take place without appreciable intravascular destruction.

In vitro effects of temperature on red blood cell deformability and membrane stability in human and various vertebrate species

2021 ◽  
pp. 1-10
Author(s):  
Adam Attila Matrai ◽  
Gabor Varga ◽  
Bence Tanczos ◽  
Barbara Barath ◽  
Adam Varga ◽  
...  
Keyword(s):  
Blood Cell ◽  
Red Blood Cell ◽  
Whole Blood ◽  
Mechanical Stability ◽  
Cell Deformability ◽  
Blood Samples ◽  
Effects Of Temperature ◽  
The Way ◽  
Rbc Deformability

BACKGROUND: The effects of temperature on micro-rheological variables have not been completely revealed yet. OBJECTIVE: To investigate micro-rheological effects of heat treatment in human, rat, dog, and porcine blood samples. METHODS: Red blood cell (RBC) - buffer suspensions were prepared and immersed in a 37, 40, and 43°C heat-controlled water bath for 10 minutes. Deformability, as well as mechanical stability of RBCs were measured in ektacytometer. These tests were also examined in whole blood samples at various temperatures, gradually between 37 and 45°C in the ektacytometer. RESULTS: RBC deformability significantly worsened in the samples treated at 40 and 43°C degrees, more expressed in human, porcine, rat, and in smaller degree in canine samples. The way of heating (incubation vs. ektacytometer temperation) and the composition of the sample (RBC-PBS suspension or whole blood) resulted in the different magnitude of RBC deformability deterioration. Heating affected RBC membrane (mechanical) stability, showing controversial alterations. CONCLUSION: Significant changes occur in RBC deformability by increasing temperature, showing inter-species differences. The magnitude of alterations is depending on the way of heating and the composition of the sample. The results may contribute to better understanding the micro-rheological deterioration in hyperthermia or fever.

scholarly journals Absence of the Adaptor Protein PEA-15 Is Associated with Altered Pattern of Th Cytokines Production by Activated CD4+ T Lymphocytes In Vitro, and Defective Red Blood Cell Alloimmune Response In Vivo

PLoS ONE ◽  
2015 ◽  
Vol 10 (8) ◽  
pp. e0136885 ◽  
Author(s):  
Stéphane Kerbrat ◽  
Benoit Vingert ◽  
Marie-Pierre Junier ◽  
Flavia Castellano ◽  
François Renault-Mihara ◽  
...  
Keyword(s):  
T Lymphocytes ◽  
Blood Cell ◽  
Red Blood Cell ◽  
Adaptor Protein

scholarly journals Effect of oxalate and malonate on red cell metabolism

Blood ◽  
1987 ◽  
Vol 70 (5) ◽  
pp. 1389-1393
Author(s):  
E Beutler ◽  
L Forman ◽  
C West
Keyword(s):  
Pyruvate Kinase ◽  
Cell Metabolism ◽  
Inhibitory Effect ◽  
Red Cells ◽  
Red Cell ◽  
Acid Analogue ◽  
2,3 Dpg

The addition of oxalate to blood stored in Citrate-phosphate-dextrose (CPD) produces a marked improvement in 2,3-diphosphoglycerate (2,3-DPG) preservation; an increase in 2,3-DPG levels can also be documented in short-term incubation studies. Oxalate is a potent in vitro inhibitor of red cell lactate dehydrogenase, monophosphoglycerate mutase, and pyruvate kinase (PK). In the presence of fructose 1,6-diphosphate the latter inhibitory effect is competitive with phospho(enol)pyruvate (PEP). Determination of the levels of intermediate compounds in red cells incubated with oxalate suggest the presence of inhibition at the PK step, indicating that this is the site of oxalate action. Apparent inhibition at the glyceraldehyde phosphate dehydrogenase step is apparently due to an increase in the NADH/NAD ratio. Oxalate had no effect on the in vivo viability of rabbit red cells stored in CPD preservatives for 21 days. Greater understanding of the toxicity of oxalate is required before it can be considered suitable as a component of preservative media, but appreciation of the mechanism by which it affects 2,3-DPG levels may be important in design of other blood additives. Malonate, the 3-carbon dicarboxylic acid analogue of oxalate late did not inhibit pyruvate kinase nor affect 2,3-DPG levels.

scholarly journals Desensitization of Adrenaline-Induced Red Blood Cell H+ Extrusion in Vitro After Chronic Exposure of Rainbow Trout to Moderate Environmental Hypoxia

1991 ◽  
Vol 156 (1) ◽  
pp. 233-248 ◽  
Author(s):  
S. THOMAS ◽  
R. KINKEAD ◽  
P. J. WALSH ◽  
C. M. WOOD ◽  
S. F. PERRY
Keyword(s):  
Rainbow Trout ◽  
Blood Cell ◽  
Red Blood Cell ◽  
Whole Blood ◽  
Inverse Correlation ◽  
Adrenergic Stimulation ◽  
Plasma Adrenaline ◽  
Arterial Blood Gas ◽  
Arterial Blood

The sensitivity of red blood cell Na+/H+ exchange to exogenous adrenaline was assessed in vitro using blood withdrawn from catheterized rainbow trout (Oncorhynchus mykiss) maintained under normoxic conditions [water PO2, (PwO2)=20.66 kPa] or after exposure to moderate hypoxia (PwO2=6.67-9.33 kPa) for 48 h, which chronically elevated plasma adrenaline, but not noradrenaline, levels. Peak changes in whole-blood extracellular pH over a 30 min period after adding 50–1000 nmoll−1 adrenaline were employed as an index of sensitivity; the blood was pre-equilibrated to simulate arterial blood gas tensions in severely hypoxic fish (PaO2=2.0 kPa, PaCO2=0.31 kPa). Blood pooled from normoxic fish displayed a dose-dependent reduction in whole-blood pH after addition of adrenaline. Blood pooled from three separate groups of hypoxic fish, however, displayed diminished sensitivity to adrenaline, ranging from complete desensitization to a 60%reduction of the response. Subsequent experiments performed on blood from individual (i.e. not pooled) normoxic or hypoxic fish demonstrated an inverse correlation between the intensity of H+ extrusion (induced by exogenous adrenaline addition) and endogenous plasma adrenaline levels at the time of blood withdrawal. However, acute increases in plasma adrenaline levels in vitro did not affect the responsiveness of the red blood cell to subsequent adrenergic stimulation. The intensity of H+ extrusion was inversely related to the PaO2in vivo between 2.67 and 10.66 kPa, and directly related to the logarithm of the endogenous plasma adrenaline level. The results suggest that desensitization of Na+/H+ exchange in chronically hypoxic fish is related to persistent elevation of levels of this catecholamine. This desensitization can be reversed in vitro as a function of time, but only when blood is maintained under sufficiently aerobic conditions.

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