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PubMed1998

The early phase of engraftment after murine blood cell transplantation is mediated by hematopoietic stem cells.

Blood cells transplantation is largely replacing bone marrow transplantation because engraftment is more rapid. This accelerated engraftment is thought to be mediated by relatively mature committed hematopoietic progenitor cells. Herein, we have used a modified rhodamine (Rho) staining procedure to identify and purify Rho+/++ (dull/bright) and Rho- (negative) subpopulations of hematopoietic progenitor cells in murine cytokine-mobilized blood. The Rho+/++ cell population contained > 99% of committed progenitor cells with in vitro colony-forming ability. The Rho- cell population contained the majority of hematopoietic stem cells with in vivo marrow repopulating ability. The rate of hematopoietic reconstitution was identical in recipients of grafts containing only purified Rho- stem cells or purified Rho- stem cells in combination with large numbers of Rho+/++ committed progenitor cells. In contrast, transplantation of 3-fold more hematopoietic stem cells resulted in accelerated reconstitution, indicating that the reconstitution rate was determined by the absolute numbers of Rho- stem cells in the graft. In addition, we observed a 5- to 8-fold reduced frequency of the subset of hematopoietic stem cells with long-term repopulating ability in cytokine-mobilized blood in comparison to steady-state bone marrow. Our results indicate that hematopoietic stem cells and not committed progenitor cells mediate early hematopoietic reconstitution after blood cell transplantation and that relative to bone marrow, the frequency of stem cells with long-term repopulating ability is reduced in mobilized blood.

Animals↗
PubMed1991

[Morphologic and functional properties of blood cells and blood rheologic parameters after plasmapheresis in patients with unstable angina pectoris].

The state of surface membrane of red blood cells and blood platelets, blood platelet adhesion to glass, red blood cell deformability, viscosity of the whole blood and plasma were studied in patients with coronary heart disease who had been subjected to blood gravitation surgery correction in the combined treatment. Normalization of all the parameters studied was recorded after inclusion of the methods of extracorporeal regulation of blood aggregation into the combined treatment. An attempt has been made to explain the mechanisms of gravitation surgery effect on the morphological and functional properties of blood cells and rheologic parameters.

Angina, Unstable↗
PubMedDate not supplied

IgA content of frozen-thawed-washed red blood cells and blood products measured by radioimmunoassay.

The IgA concentration of various blood products was examined by a radioimmunoassay, and the lowest IgA concentrations were seen in frozen-thawed-washed red blood cells. The routine Regional Blood Transfusion Centre preparation contained a mean IgA concentration of 0.117 mg/l (n=21) in the frozen-thawed-washed red blood cell supernatant fluid. Further washing of frozen-thawed-washed red blood cells with saline reduced supernatant IgA concentrations to below the limit of IgA detection (less than 0.003 mg/l). Ordinary washed red blood cells and other blood plasma fractions contained substantial quantitites of IgA, supporting the view that frozen-thawed-washed red blood cells should be used for transfusion of patients at risk of developing reactions to IgA, and the need for caution in the use of other blood products in such patients.

Anaphylaxis↗
PubMedDate not supplied

Comparison of methods to wash liquid-stored red blood cells and red blood cells frozen with high or low concentrations of glycerol.

The efficiency of washing liquid-stored red blood cells and red blood cells frozen with high or low glycerol concentrations was evaluated by measuring the recovery of red blood cells in vitro, supernatant hemoglobin, extracellular potassium and red blood cell potassium levels, supernatant osmolality, residual 125I albumin, glycerol, hypoxanthine, and di-2-ethylhexyl phthalate (DEHP) levels. Four commercial washing systems were studied, three which used sodium chloride solutions with serial or continuous-flow centrifugation and one which used sugar solutions and dilution/agglomeration. Washing was most efficient using sodium chloride solutions in the IBM Blood Processor, an automated serial centrifugation procedure and in the Fenwal Elutramatic, a continuous-flow centrifugation procedure. Less efficient washing was achieved in the Haemonetics Processor 15, a continuous-flow centrifugation procedure and the least efficient washing occurred using the original and modified dilution/agglomeration procedures. To achieve the most efficient washing, three principles must be utilized: concentration of the red blood cells to hematocrit values of 90 per cent, prior to washing or freezing. Liquid-stored red blood cells concentrated to hematocrit values of 90V per cent should be diluted with hypertonic sodium chloride solutions prior to recovery and washing. Red blood cells containing 20 per cent or 40 per cent W/V glycerol should be diluted with hypertonic sodium chloride solutions before recovery and washing. Finally, on-line dilution should be achieved in the washing systems that use continuous-flow centrifugation.

Blood Preservation↗
PubMedDate not supplied

Squirrel monkey (Saimiri sciureus) B lymphocytes: secretion of IgG directed to Plasmodium falciparum antigens, by primed blood B lymphocytes restimulated in vitro with parasitized red blood cells.

Blood B lymphocytes obtained from Plasmodium falciparum-immune Saimiri monkeys were assayed for their in vitro differentiation in immunoglobulin-secreting cells upon restimulation with P. falciparum-parasitized Saimiri red blood cells. Selected culture conditions enabled appropriately stimulated blood B cells to secrete 3F11/G10+ IgG, detected in the supernatants by means of a dot immunobinding assay. Primed blood B lymphocytes from P. falciparum-immune Saimiri monkeys were thus able to secrete IgG when restimulated by parasitized red blood cells in the presence of T cell- and monocyte-derived cytokines (recombinant human cytokines). These primed blood B cells, which were able to differentiate, were shown to secrete antibodies reactive with P. falciparum-infected red blood cells, as detected by means of an indirect immunofluorescence assay, and reactive with P. falciparum-infected red blood cell extracts, as detected by means of Western blot analysis. Furthermore, due to the possibility of discriminating between IgG subtypes in the squirrel monkey (3F11/G10+::3A2/G6+ IgG [associated with protection against the blood stages of P. falciparum] vs. 3F11/G10+::3E4/H8+ IgG [usually not functionally associated with protection]), we have attempted to estimate the respective proportions of each IgG subtype. In defined culture conditions, Saimiri monkey blood B cells preferentially secrete 3F11/G10+::3E4/H8+ IgG in response to parasitized red blood cells. We therefore discuss the conditions that would render this assay suitable for the selection, among P. falciparum blood stage antigens, of those that have major B-cell epitopes.

Animals↗
PubMed1992

Mobilization of circulating haemopoietic cells (blood stem cells) by GM-CSF in patients with malignancies.

Autologous reinfusion of circulating haemopoietic progenitor and stem cells (blood stem cell transplantation) has emerged as an alternative to autologous bone marrow transplantation in a variety of malignant diseases. Major obstacles associated with harvest of blood stem cells by leukapheresis are: 1. relatively high costs, and 2. discomfort caused to the patient, as generally five to ten settings of leukapheresis are necessary to harvest a number of blood stem cells sufficient for haemopoietic restitution following myeloablative therapy. GM-CSF recently has been shown to effectively increase circulating haemopoietic cells, when given subsequent to even highly-toxic therapy. This report summarizes our data on mobilization of blood stem cells by GM-CSF cells in multiple myeloma patients.

Combined Modality Therapy↗
PubMed1988

NMR water-proton spin-lattice relaxation time of human red blood cells and red blood cell suspensions.

NMR water-proton spin-lattice relaxation times were studied as probes of water structure in human red blood cells and red blood cell suspensions. Normal saline had a relaxation time of about 3000 ms while packed red blood cells had a relaxation time of about 500 ms. The relaxation time of a red cell suspension at 50% hematocrit was about 750 ms showing that surface charges and polar groups of the red cell membrane effectively structure extracellular water. Incubation of red cells in hypotonic saline increases relaxation time whereas hypertonic saline decreases relaxation time. Relaxation times varied independently of mean corpuscular volume and mean corpuscular hemoglobin concentration in a sample population. Studies with lysates and resealed membrane ghosts show that hemoglobin is very effective in lowering water-proton relaxation time whereas resealed membrane ghosts in the absence of hemoglobin are less effective than intact red cells.

Adult↗
PubMed1978

Blood volume measurements in gopher snakes, using autologous 51Cr-labeled red blood cells.

Blood volume determinations were performed in 5 anesthetized gopher snakes (Pituophis melanoleucus catenifer) by means of a 51Cr-labeled red blood cell (RBC) method. The mean blood volume was 52.8 ml/kg of body weight (+/- 6.21 SE). Previous blood volume measurements have not been reported for this species. The RBC survival rate was estimated to be greater than 660 days. The RBC survival rate is long, but it cannot be determined accurately by this method.

Animals↗
PubMed1994

Differential localization of myosin-II isozymes in human cultured cells and blood cells.

We used purified polyclonal antibodies to human cytoplasmic myosin-IIA and myosin-IIB directly labeled with fluorescent dyes to localize these myosin-II isozymes in HeLa cells, melanoma cells and blood cells. Both antibodies react strongly with myosin-II isozymes in HeLa cells, melanoma cells and blood eosinophils, but only anti-myosin-IIA antibodies stain platelets, lymphocytes, neutrophils and monocytes in smears of human blood. Both antibodies stain small spots along the stress fibers of interphase HeLa cells and melanoma cells, but double staining revealed that the detailed distributions of myosin-IIA and myosin-IIB differ. A low concentration of diffuse myosin-IIB is present in the cortex, both in lamellar regions around the periphery of the cell and over the free surface. Myosin-IIB is also concentrated in spots along perinuclear stress fibers. Myosin-IIA is absent from the cortex but is concentrated in spots along stress fibers located near the basal surface of cultured cells. This population of peripheral stress fibers is highly enriched in myosin-IIA relative to myosin-IIB, but both are found together in centrally located stress fibers. In prophase and metaphase both isozymes are concentrated in the cortex in small spots less than 04.micron in size, similar to those in stress fibers. As the chromosomes begin the separate at anaphase, most of the myosin-II spots become concentrated in the outer 0.7 micron of the equatorial cortex in 100% of cells. This concentration of myosin-II isozymes in the cleavage furrow is maintained until the daughter cells separate. The superimposition of these small spots concentrated in the cleavage furrow produces the intense, uniform staining observed in conventional micrographs of whole cells.

Actins↗
PubMed2006

Do placental sections accurately reflect umbilical cord nucleated red blood cell and white blood cell differential counts?

OBJECTIVE: Elevated levels of umbilical cord nucleated red blood cells (nRBCs) have been used to assess in utero hypoxia. Although the umbilical nRBC value has been the 'gold standard', umbilical blood may not be obtained at delivery. We determined if the levels of nRBCs and white blood cell (WBC) counts in fixed placental sections might serve as a proxy for cord blood values. STUDY DESIGN: Umbilical blood and placenta were collected from 25 deliveries at Harbor-UCLA Medical Center. Umbilical blood and placental sections were analyzed for nRBCs (per 100 WBC) and WBC differential, and compared with the t-test or the Mann-Whitney rank sum test. RESULTS: nRBC counts were equivalent in umbilical cord and placental sections (5 vs. 4/100 WBC). Umbilical lymphocyte and polymorphonuclear leukocyte (PMN) counts were normally distributed, averaging 35 +/- 9 and 56 +/- 2/100 WBC, respectively. Placental lymphocyte (33 +/- 2/100 WBC) and PMN (60 +/- 2/100 WBC) counts were equivalent to cord blood values. CONCLUSION: WBC differentials and nRBC counts are equivalent in umbilical cord blood and processed placental pathology sections. For infants in whom cord blood cell counts are desired though umbilical cord samples are unavailable, fixed placental sections may serve as a proxy.

Adult↗
PubMed1999

Quantitative PCR for counting residual white blood cells in blood products.

Leukocyte depleted blood components are frequently used to reduce alloimmunization and the risk of transfusion transmitted infection. Counting residual white blood cells in filtered blood products requires sensitive and reliable techniques. After separation of white blood cells from 500 microliters of 20 non-filtered and 54 filtered blood products we used polymerase chain reaction (PCR) and fluorimetric detection for the quantification of genomic DNA. The results were compared with results from Nageotte chamber counting. The accurate limit of detection of PCR was determined at 1 WBC/microliter (intra-assay coefficient of variation: 16.3%). PCR correlated well with Nageotte chamber counts (r = 0.77, p < 0.001, n = 74). Concordant results were obtained in 51 filtered and 20 non-filtered blood products. Discrepant results were obtained in 3 filtered whole blood units: In these blood products > 12 WBC/microliters were counted in Nageotte chamber and PCR gave a negative result. After component preparation fresh-frozen plasma and red cell concentrates of these units contained < 1 WBC/microliter using both methods. In conclusion we describe a quantitative PCR method which had about the same sensitivity and specificity as Nageotte chamber testing. However, PCR is more laborious than the standard method. As well, as reliable PCR testing requires expensive instruments and staff experienced in molecular biology, the standard method is more cost effective.

Fluorometry↗
PubMed1986

Influence of dystocia on white blood cell and blood neutrophil counts in mares.

A retrospective study was done on total white blood cell (WBC) and blood neutrophil counts of 41 mares referred to one of two veterinary hospitals for correction of dystocia. The mares were 2 to 19 years of age and included draft, light, and pony breeds. The WBC and neutrophil counts were performed at varying intervals from time of admission to 10 d after delivery of the feti. Retrospective analyses of WBC and neutrophil counts from 10 normal foaling mares from two Pennsylvania breeding farms (Thoroughbred and Trakehner) and from 14 normal foaling pony mares were done as controls. Mean WBC (10446 +/- 2296 cells/mul) and neutrophil (6850 +/- 2136 cells/mul) counts on the day of delivery in mares with normal parturition were slightly elevated over values reported as normal in the literature. The mean blood cell counts gradually declined to 6124 +/- 1255 WBC/mul and 3692 +/- 409 neutrophils/mul on Day 2 postpartum and returned to normal baseline values by Day 3 postpartum (8868 +/- 2693 WBC/mul, 4298 +/- 1966 neutrophils/mul). No toxic neutrophils were present in mares with normal delivery. Mean WBC (11346 +/- 3298 cells/mul) was elevated on the day of delivery in mares with dystocia as a result of neutrophilia with a left shift (9297 +/- 3298 neutrophils/mul). An apparently faster decline occurred in WBC and neutrophil counts of mares with dystocia than in mares with normal delivery, until a marked leukopenia (3905 +/- 1292 WBC/mul) and neutropenia (1570 +/- 1340 neutrophils/mul) occurred on Day 3 postpartum. The leukopenia and neutropenia persisted until Day 5 postpartum. Toxic neutrophils were present in several mares with dystocia.

Journal Article↗
PubMed2002

Automated counting of nucleated red blood cells in blood samples of newborns.

Nucleated red blood cells (NRBC) in blood samples interfere with the white blood cell (WBC) count on many types of automated haematology analysers. This makes it necessary to correct the WBC count by counting NRBC microscopically. This report describes the evaluation of two analysers, the Cell-Dyn 4000 and the Sysmex XE-2100, which use new techniques to recognize and enumerate NRBC. We conclude that both the Cell-Dyn 4000 and the Sysmex XE-2100 give an accurate WBC count in the presence of NRBC. Furthermore, they can enumerate NRBC correctly when compared with microscopic observation.

Artifacts↗
PubMed1978

Effect of centrifugation and subsequent storage on red blood cells.

Blood drawn into CPD solution from 33 normal donors was divided into four groups: (I) centrifuged (at 5,000 g for 7 min) after 7 days of storage, (II) centrifuged after 14 days storage, (III) centrifuged after 21 days storage and (IV) uncentrifuged. After 21 days of storage, aliquots of all units were labeled with chromium-51, reinjected into the donor from which they were drawn and erythrocyte survival was measured. Red blood cell recovery and survival for all four groups was essentially the same; 24-hour recovery was 85%; T 1/2 was 28.2--31.6 days. Our results suggest that blood can be centrifuged and stored at any time during its 21-day shelf life without detrimental effect on erythrocyte survival.

Adult↗
PubMed1997

[Advances in molecular biological investigation of blood group-active substances on the human red blood cell].

Blood group antigens on the human red blood cell are originally of serological significance. Recent advances in molecular biology and genetics have greatly increased our knowledge of the chemical structures, functions, and genetic backgrounds of these antigens. Carbohydrate antigens are widely expressed in various tissues, whereas protein(polypeptide) antigens are generally specific to the surface molecules of erythroid cells, suggesting their possible roles in membrane structure and function (transporters, receptors, adhesion molecules, and enzymes). This paper reviewed the recent topics with special regard to clinical significance including blood transfusion, and discussed.

ABO Blood-Group System↗
PubMed1977

Chimeric drift in allophenic mice: analysis of changes in red blood cell and white blood cell populations in C57Bl/6 in equilibrium (A X SJL)F1, C57Bl/6 in equilibrium (CBA X CBA/H-T6)F1, and C57Bl/6 in equilibrium DBA/1 mice.

Forty-seven allophenic mice of three different types (C57BL/6 in equilibrium (A X SJL), C57BL/6 in equilibrium (CBA X CBA/H-T6), and C57BL/6 in equilibrium DBA/1) were analyzed for changes in their peripheral white blood cell composition and hemoglobin composition with age. It was found that 10 of the 47 mice showed significant changes termed "chimeric drift" in one or the other or both of these parameters. These 10 mice were classified as unstable chimeras, as opposed to the 37 stable chimeras, which showed no apparent chimeric drift. There was an excellent correlation of peripheral white blood cell and hemoglobin compositions of the stable chimeras. However, the unstable chimeras showed little or no correlation of these two markers. Possible mechanisms of chimeric drift are discussed.

Aging↗
PubMed2005

Basic investigation of the lectin method for separation and recovery of nucleated red blood cells in maternal blood, and a study into the frequency of nucleated red blood cells in fetomaternal disorders.

We previously reported the separation and recovery of nucleated red blood cells (NRBCs) in maternal blood using the lectin method. In the present study, we verified the lectin method and investigated the appearance of NRBCs during pregnancy. For the concentration of lectin soy bean agglutinin, 7 mL of maternal peripheral blood was collected from 20 subjects, and the relative fluorescence intensity was measured using flowcytometry; 50 mg/mL, used in previous studies, was the optimal concentration. The number of cells recovered at each step of the lectin method was also investigated by FACS using fluorescence-labeled CD11a and CD33, and the results showed the usefulness of the method. Next, 7 mL of maternal peripheral blood was collected from 292 women with a normal single pregnancy (389 specimens), and NRBCs were separated and recovered using the lectin method. NRBCs slightly increased over the course of pregnancy (y = 4.29x + 5.03, r2 = 0.11). When blood was collected multiple times in the same subjects, NRBCs increased in 63 of 77 subjects (83.1%, percent change: 2.4 +/- 19.0). No NRBCs were recovered in 17 subjects (4.7%). Regarding the relationship between fetomaternal disorders and the frequency of NRBCs, 89.4 +/- 92.6 cells appeared per 10 mL of maternal blood in the normal group, but NRBCs increased in patients with 18 trisomy, placenta previa, pre-eclampsia, intrauterine fetal death, and 21 trisomy. NRBC examination may play an assisting role not only in fetal diagnosis but also in fetomaternal diagnosis.

Cell Nucleus↗
PubMed2005

Cremophor EL releases cyclosporin A adsorbed on blood cells and blood vessels, and increases apparent plasma concentration of cyclosporin A.

We examined the influence of cremophor EL (crEL) on the disposition kinetics of CyA in rats. A dose of 10mg/kg of CyA in a volume of 750 microL containing 4.3, 16 or 30% concentration of crEL was intravenously administered over 1 min to rats. The values of distribution volume at the steady-state (Vd(ss)) and total clearance (CL(tot)) of CyA in the presence of increasing amounts of crEL were decreased to about 1/3-1/5 of those with 4.3% crEL, in a crEL concentration-dependent manner. The values of blood to plasma concentration ratio (RBP) and the apparent tissue to plasma concentration ratio (K(p,app)) of CyA with 30% crEL were both only about 1/2 of those of CyA with 4.3% crEL. Next, rats were intravenously given 30% crEL solution at 30 min after an intravenous administration of CyA (10 mg/kg) with 4.3% crEL. Subsequently, the blood and plasma concentrations of CyA rose significantly to 2.4 and 4.7 times those seen when i.v. 30% crEL was not given, respectively. In an in vitro study, we found that the uptake of CyA by red blood cells is inhibited by crEL, and that CyA adsorbed on the inner surface of blood vessels after the administration of CyA is released by crEL. The disposition kinetics of CyA is altered by i.v. administration in combination with the surfactant vehicle crEL, in a crEL concentration-dependent manner.

Adsorption↗