Evaluation of an experimental filter designed for improving the quality of red blood cells (RBCs) during storage by simultaneously removing white blood cells and immunomodulators and improving RBC viscoelasticity and Band 3 proteins.

BACKGROUND: Storage age of red blood cells (RBCs) has been reported to be associated with increased mortality and morbidity. During storage, RBCs undergo changes in biochemical and functional properties. Stored RBCs may also contain white blood cells (WBCs), activated platelets (PLTs), cytokines, immunoglobulin, and other bioactive proteins. Transfusion of these bioactive proteins and cells with RBCs has the potential to cause serious adverse effects. We evaluated the performances of an experimental filter (EF) designed to remove immunoglobulins, cytokines, and other bioactive proteins in RBCs. STUDY DESIGN AND METHODS: Sixteen sets, each containing 3 units of ABO-identical RBCs in AS-3 were obtained from a blood bank. Three units of RBCs were combined together and then split into three equal aliquots, A, B, and C. Unit A was unfiltered while Units B and C were filtered with a leukoreduction filter and the EF, respectively. All the units were stored at 4°C in a blood bank refrigerator for 42 days. We measured RBC viscoelasticity, hemolysis, RBC adenosine triphosphate, Band 3 proteins, cytokines, PLTs, WBCs, and immunoglobulin before and after filtration and on Days 21 and 42 of storage. Data were analyzed by repeated-measures analysis of variance with Newman-Keuls multiple comparison test. RESULTS: The EF significantly (p<0.05) reduced the levels of immunoglobulin (control IgG, 2.184 ± 1.918 mg/mL; BPF4, 2.216 ± 1.956 mg/mL; and EF, 0.363 ± 0.391 mg/mL), PLTs, cytokines, and improved viscoelastic properties when compared to either control or leukoreduced RBCs. CONCLUSION: The EF achieved lower levels of WBCs, improved viscoelastic properties, and reduced levels of immunoglobulins and cytokines but significance will require clinical evaluation.

Antibodies to the HLA-A2 antigen prime neutrophils and serve as the second event in an in vitro model of transfusion-related acute lung injury.

BACKGROUND: Transfusion-related acute lung injury (TRALI) is the most common cause of transfusion-related mortality and has been linked to the infusion of donor antibodies directed against recipient HLA class I antigens. We hypothesize that antibodies against HLA class I antigens bind to the antigens on the neutrophil (PMN) surface and induce priming and PMN cytotoxicity as the second event in a two-event in vitro model of PMN-mediated cytotoxicity. METHODS: Isolated PMNs from HLA-A2 homozygotes, heterozygotes and null donors were incubated with a monoclonal antibody to HLA-A2 and a human polyclonal IgG to HLA-A2 and priming of the oxidase was measured. The monoclonal antibodies and PMNs from these three groups were then used in a two-event model of PMN cytotoxicity. RESULTS: The antibodies to HLA-A2 both primed PMNs from HLA-A2 homozygotes but not from heterozygotes or nulls. Antibodies to HLA-A2 also served as the second event in a two-event model to induce PMN cytotoxicity of HLA-A2 homozygous PMNs. CONCLUSION: Antibodies to HLA class I antigens may directly prime/activate PMNs through the ligation of the antigen on the cell surface, and the antigen density appears to be important for these changes in PMN physiology.

A simple filtration system for red blood cell depletion and volume reduction in routine processing of human umbilical cord blood.

BACKGROUND AND OBJECTIVES: Currently, stem cells and other progenitor cells are obtained from human umbilical cord blood (HUCB) using a variety of methods that are designed primarily for red blood cell depletion and volume reduction prior to freezing and storage. Some of these methods are very cumbersome and involve several steps that may result in significant cell loss. Therefore, processes that minimize the loss of haematopoietic stem and progenitor cells (HSPC) remains very critical. In the present study, we describe a simple filtration process for achieving both volume reduction and red blood cell depletion in a 'closed sterile system' with significant recovery of viable HSPC. MATERIALS AND METHODS: About 80-100 ml of HUCB were collected into citrate-phosphate-dextrose-adenine 1 anticoagulant. Each HUCB was divided into 25-70-ml aliquots and then either diluted with isotonic saline or filtered without any prior dilution with an experimental Red Cell Volume Reduction System (RCVRS). The HSPCs were recovered by retrograde rinsing of the filter with an isotonic stem cell recovery solution. The viability, colony forming properties, leucocytes and CD34+ cells recoveries were determined. RESULTS: The mean volume of the HUCB before processing was reduced from 43.9 +/- 7.9 ml to 11.8 +/- 0.7 ml (n = 55) with red blood cell depletion of 85.2 +/- 3.7%. Diluting the HUCB with isotonic saline prior to processing with RCVRS increased the red blood cell depletion to 91.9 +/- 3.0% (n = 7) without any significant loss in viability or cell recovery. The mean viability of the RCVRS-processed HUCB was not significantly different from the control unprocessed blood (96.60 +/- 1.90 vs. 96.63 +/- 2.12%; P > 0.05). The mean recoveries of the CD34+ and the haematopoietic clonogenic progenitor cells with the filter were 83.9 +/- 26.8 (n = 40) and 99.9 +/- 27.9% (n = 35), respectively. CONCLUSION: The present results show that the RCVRS provides a simple and easy-to-use process for obtaining red blood cell depletion and volume reduction of HUCB with good cell viability and recoveries.

Pall leukotrap affinity prion-reduction filter removes exogenous infectious prions and endogenous infectivity from red cell concentrates.

BACKGROUND AND OBJECTIVES: Three recent probable cases of transmission of a variant of human Creutzfeldt-Jakob disease (vCJD) through blood transfusion suggest that the disease can be transmitted through transfusion of blood components from presymptomatic blood donors. In this study, we investigated the performance of a new filter for reducing the levels of infectious prions (PrP(Sc)) from red cell concentrates (RCC). MATERIALS AND METHODS: Endogenous Infectivity: A pool of 500 ml of whole blood was collected from 263K-strain scrapie-infected hamsters into an anticoagulant, processed into non-leucoreduced RCC (NL-RCC), and then passed through a prion-reduction filter. Pre- and postfiltration samples were tested for PrP(Sc) by Western blot and infectivity by inoculation of healthy hamsters. Results of the endogenous infectivity study after 200 days post-inoculation are discussed. Exogenous (Spiking) Study: Scrapie-infected hamster brain homogenates containing PrP(Sc) were added to human RCC and then filtered. Levels of PrP(Sc) were determined by Western blot assay. The effect of prior leucodepletion of 'spiked' RCC on PrP(Sc) removal by the prion-removal filter was also assessed. RESULTS: In the endogenous infectivity study, at 200-day observation time, the prefiltered RCC transmitted disease to six of the 187 hamsters, whereas the filtered RCC did not transmit disease to any of 413 animals, P = 0.001. The prion filter also significantly reduced the concentration of leucocytes in the RCC by about 4 logs, P < 0.05. In the exogenous (spiking) study, the level of PrP(res) was significantly reduced in RCC P < 0.05. Prior leucodepletion of the RCC with a leucoreduction filter did not significantly reduce the concentration of exogenously spiked PrP(Sc), P > 0.05. CONCLUSION: The use of this new prion-reduction filter should reduce the risk of vCJD transmission through transfusion of RCC, the most widely transfused blood component.

Prestorage leukoreduction and low-temperature filtration reduce hemolysis of stored red cell concentrates.

BACKGROUND: Universal prestorage leukoreduction in Canada created the perception that stored red cells (RBCs) are more hemolyzed than their unfiltered predecessors. A pool-split design tested the effects of leukoreduction on hemolysis of stored RBCs. STUDY DESIGN AND METHODS: Two ABO-matched units were pooled, divided, and then processed into leukoreduced (LR) and nonleukoreduced (NLR) units with the Pall LT-WB or RC-PL systems and sampled during standard processing and storage for testing of sterility, counts, hemolysis, and osmotic fragility. RESULTS: Room temperature (RT) filtration of 10 pairs of LT-WB-LR and -NLR units showed significantly different percentage of hemolysis (0.39%) and osmotic fragility (0.643%) at 42 days. Cold-stored and -filtered units (2 days at 4 degrees C before processing) were less hemolyzed, but showed a similar proportional decrease of hemolysis in LR units (0.13% vs. 0.25% at 42 days). RBCs from RC-PL systems showed the lowest hemolysis although there was a filtration effect (0.05% vs. 0.12%, 42 days). Osmotic fragility paralleled hemolysis. Segment samples gave inaccurate results. Two-day prefiltration cold storage reduced hemolysis from 0.36 to 0.07 percent (42 days, p < 0.001). RT-LR hemolysis became significantly higher by Day 10 and 4 degrees C LR by Day 12. NLR units showed hemolysis by Day 7. LR units filtered cold were less hemolyzed (p < 0.05) than RT-LR but osmotic fragility was unchanged. CONCLUSIONS: LR-RBCs prepared by any of three methods (LT-WB, RT or cold; RC-PL), filtered at 4 degrees C, were less hemolyzed during storage than nonfiltered concentrates: 4 degrees C leukoreduction is beneficial for RBCs and does not cause hemolysis or enhance fragility.

White cell subsets in apheresis and filtered platelet concentrates.

BACKGROUND: White cell (WBC)-reduced platelet concentrates (PCs) are defined by their absolute WBC count, a criterion which provides no information regarding the various WBC subsets contained in the PC. These heterogeneous cells are known to mediate different physiologic and pathophysiologic functions and account for distinct adverse transfusion responses. This study describes a method which allows the detection and quantification of these subsets and characterizes their presence in a variety of platelet components. STUDY DESIGN AND METHODS: Random-donor pooled PCs (RD PCs) and single-donor apheresis PCs (SD PCs) were studied. RD PCs consisting of 6 units of 2- to 3-day old PCs were randomly assigned to be filtered with one of four WBC-reduction filters from three different manufacturers (n=34). The residual WBCs were pelleted by centrifugation and isolated on a density gradient. The various WBC subsets were quantified by flow cytometry in unfiltered and filtered PCs using fluorescence and two-angle light scatter. SD PCs obtained with two manufacturer's systems and three processing protocols (n=30) were studied in like manner. RESULTS: WBC counts for non-WBC-reduced PCs averaged 3 x 10(8) in RD PCs and ranged from 8.6 to 9.6 x 10(6) per SD PC. Residual WBC counts in filtered PCs ranged from 2.3 x 10(4) to 2.2 x 10(5) and those in WBC-reduced SD PCs averaged 2.2 x 10(5) per unit. The data demonstrate significant phenotypic differences among PCs produced with various procedures. All SD PCs and two of four filtered RD PCs contained five WBC populations including granulocytes and monocytes, while RD PCs filtered with the remaining manufacturer's devices contained only lymphocytes. CONCLUSION: The data confirm that distinct phenotypic differences exist among PCs prepared with different devices and/or procedures. It is suggested that as for non-generic pharmaceuticals, the clinical benefits of these various PCs should be individually proved.

Survival of lyophilized and reconstituted human red blood cells in vivo.

To assess the viability of human red blood cells that have been lyophilized and reconstituted to the hydrated state, we phlebotomized a unit of whole blood from six healthy male volunteers. Their packed red blood cells were lyophilized at -40 degrees C and stored at 4 degrees C. Upon rehydration, recovery of erythrocytes was 85.2 +/- 2.79%. Aliquots of 20 ml were labeled with 51Cr and re-infused into the original donors for red cell survival studies. The red cells retained ABO and Rh identity upon rehydration. There were no adverse clinical affects of re-infusion. The half time of 51Cr disappearance from the circulation was 31 +/- 8.19 days, and there was no evidence of significant splenic sequestration on the day of reinfusion. Red cell indices of the rehydrated erythrocytes were normal, oxyhemoglobin content was 98.58 +/- 1.46%, and P50 was 27.25 +/- 1.84 mmHg. Although deformability was slightly decreased, the osmotic fragility and filterability of the red cells were normal. These data demonstrate that human erythrocytes can be lyophilized and reconstituted to the hydrated state and survive normally in the circulation. Metabolic, osmotic, hematological and rheological function remains intact.

Refrigerated storage of lyophilized and rehydrated, lyophilized human red cells.

Human red cells (RBCs) were collected in CPDA-1 and then freeze-dried in lyoprotective solution. The lyophilized RBCs were then stored at -20 degrees C for 7 days. At the end of the storage period, the lyophilized RBCs were rehydrated and washed in dextrose saline. The washed, reconstituted, lyophilized RBCs were resuspended in final wash solutions of ADSOL, CPDA-1, or a special additive solution containing glucose, citrate, phosphate, adenine, and mannitol, and then they were stored at 4 degrees C for an additional 7 days. The main purpose of this study was to determine whether human RBCs can be lyophilized in such a manner that normal metabolic, rheologic, and cellular properties are maintained during rehydration and subsequent storage in standard blood bank preservative solutions. Our results show that reconstituted, lyophilized RBCs maintained levels of ATP, 2,3 DPG, lactate, and cellular properties that are equal to or better than those in control nonlyophilized RBCs stored for a comparable period in CPDA-1. Reconstituted, lyophilized RBCs stored at 4 degrees C after rehydration also show better maintenance of ATP, 2,3 DPG, and lactate than do control RBCs stored in the same preservative solutions for comparable periods.

Effects of normal and sickle erythrocytes on prostacyclin release by perfused human umbilical cord veins.

We compared the effects of normal (AA) and sickle (SS) erythrocytes (RBC) on endothelial cell release of prostacyclin by perfused human umbilical cord veins. Two equal-length segments of fresh umbilical cords were perfused first with serum-free Dulbecco's modified Eagle's medium (DMEM) to establish the basal prostacyclin production rate for each segment; then one segment was perfused with SS RBC and/or plasma, while the other segment was simultaneously perfused with AA RBC and/or plasma. Aliquots of perfusate were removed at intervals for measurement of the stable prostacyclin metabolite 6-keto-prostaglandin F1 alpha (6-keto-PGF). Basal prostacyclin production by segments from the same cord was very similar, but it varied considerably among segments from different cords. Therefore, the ratio of prostacyclin release with RBC and/or plasma to basal prostacyclin release for each segment was used to compare prostacyclin release among segments from different cords. Mean prostacyclin release was significantly higher from segments perfused with SS RBC in autologous plasma than from segments perfused with AA RBC in autologous plasma at 15, 30, and 60 min. However, no significant differences in mean prostacyclin production were observed between segments perfused with SS vs. AA RBC in DMEM or between segments perfused with SS vs. AA plasma alone. No significant correlations were observed between prostacyclin production and either the viscosity of SS and AA RBC in autologous plasma or DMEM or the adhesiveness of SS and AA RBC to cultured human umbilical vein endothelial cells. We conclude that SS RBC in autologous plasma cause increased prostacyclin release from perfused human umbilical cord veins. The perfused human umbilical cord vein system may be a useful model for comparing the response of vascular endothelium to SS and AA RBC and plasma under controlled-flow conditions.

Preservation of metabolic activity in lyophilized human erythrocytes.

Normal human erythrocytes (RBC) were freeze-dried under conditions that caused minimal modification in normal RBC metabolic activities. Because of the known effects of long-term storage on metabolic activities, we studied the effects of our lyophilization process on RBC metabolism. Of all the metabolic enzymes studied, only triosephosphate isomerase (D-glyceraldehyde-3-phosphate ketol-isomerase, EC 5.3.1.1), enolase (2-phospho-D-glyceratehydro-lyase, EC 4.2.1.11), and pyruvate kinase (ATP:pyruvate O2-phosphotransferase, EC 2.7.1.40) were decreased when compared with fresh control nonlyophilized RBC. The activities of these enzymes did not differ significantly from those of blood bank RBC. Concentrations of high-energy intermediates, ATP, and 2,3-diphosphoglycerate, along with lactate and ATP production were decreased in lyophilized RBC. No enzymes of the pentose phosphate shunt were altered during lyophilization. In addition, our data show that lyophilized RBC possess an intact capacity to (i) synthesize adenine nucleotides and (ii) reduce MetHb to Hb and, thus, maintain the Hb in a functional physiologic state similar to fresh nonlyophilized RBC. The present study demonstrates the possibility of lyophilizing RBC in a manner that maintains normal metabolic and enzymatic function upon rehydration.

Red cell aggregation and viscoelasticity of blood from seals, swine and man.

RBC aggregation and viscoelasticity parameters were determined for 40% suspensions of washed cells in autologous plasma from elephant seals (ES), Mirounga angustirostris, ringed seals (RS), Phoca hispida, and swine, (SS), Sus scrofa. Interspecific comparisons including human (HS) blood data revealed unusual rheological properties of seal blood relative to that from pigs or man: 1) RBC aggregation extent, rate and sedimentation were lower for seals (AI = 0, ZSR = .40, ESR = 0 for RS blood) relative to humans; 2) Viscous (n') and elastic (n") components of complex viscosity (OCRD) were lower for both seal species relative to SS blood, but only at shear rates less than or equal to 10 sec-1 (P less than 0.05), while n"/n' ratios for RS blood were lower than HS blood at all shear rates (P less than 0.01); 3) Blood viscosity measurements for RS and SS blood from rotational viscometry (Contraves) were consistent with OCRD data; 4) Seal plasma fibrinogen levels were low compared to pigs or humans (RS fibrinogen = -43% v. HS and -57% v. SS; ES fibrinogen = -58% v. HS and -69% v. SS). Electrophoretic mobility of RS red cells was +25% relative to those of humans. These results demonstrate differences in hemorheological indices among mammalian species and suggest the value of comparative rheologic studies.

Effect of procaine hydrochloride on the electrophoretic mobility of human red blood cells.

The interaction of cationic anesthetics with biological membranes and the resulting alterations of membrane electrokinetic properties continue to be of current interest. The present study was designed to examine the effects of procaine hydrochloride (PRHCL) on the mobility of human red blood cells (RBC); electrophoretic measurements were made on RBC suspended in phosphate-buffered saline (PBS, pH = 5.0, 7.4, or 9.2), autologous plasma or 3 g% dextran T70/PBS (pH = 7.4), with PRHCL concentrations from 8 x 10(-6) to 8 x 10(-2) M. Low concentrations of PRHCL (8 x 10(-5)-8 x 10(-3) M) significantly (p less than 0.001) increased RBC mobility, with a maximal increase of 8.2% at 8 x 10(-4) M. Conversely, a higher PRHCL concentration (8 x 10(-2) M significantly (p less than 0.001) decreased RBC mobility. Both glutaraldehyde fixation and lipid extraction abolished any PRHCL-induced increase in RBC mobility; the observed increases in mobility for normal cells are, thus, consistent with a mechanism based on expansion of the RBC membrane glycocalyx. Microelectrophoretic methods were also used to study the effect of PRHCL (8 x 10(-4) and 8 x 10(-2) M) on RBC membrane calcium binding, with the results indicating that PRHCL competes with calcium for neuraminate binding sites. We conclude that the observed changes in RBC electrokinetic properties reflect incorporation of PRHCL into the RBC membrane; such changes may be of importance in modulating cell-cell interactions.

Increased circulating endothelial cells in sickle cell crisis.

To determine whether increased numbers of circulating endothelial cells, a possible indicator of endothelial injury, are present in subjects with sickle cell disease, we measured circulating endothelial cells in 30 normal subjects and in 23 subjects with sickle cell anemia. Mean circulating endothelial cells were significantly higher (P less than 0.025) in the sickle cell subjects than in the normal subjects. Circulating endothelial cells were significantly higher than normal in 10 sickle cell subjects studied during painful crisis (P less than 0.01) but not in 13 sickle cell subjects studied while in the steady state. To control for the known stimulatory effect of cigarette smoking on circulating endothelial cells, we analyzed the results for smokers and nonsmokers separately. Mean circulating endothelial cells were not significantly higher in sickle cell subjects who smoked (n = 10) than in normal subjects who smoked (n = 8), but were significantly higher (P less than 0.05) in sickle cell nonsmokers (n = 13) than in normal nonsmokers (n = 22). Among nonsmoking sickle cell subjects, mean circulating endothelial cells were significantly higher than normal (P less than 0.01) during painful crisis (n = 7), but not in the steady state (n = 6). We conclude that circulating endothelial cells are significantly increased in sickle cell crisis, and may indicate the occurrence of acute endothelial injury during episodes of microvascular occlusion.

Effect of procaine hydrochloride on the aggregation behavior and suspension viscoelasticity of human red blood cells.

The present study examined the effects of procaine hydrochloride (PRHCL), a cationic local anesthetic, on the aggregation behavior of human red blood cells (RBC); the effects of PRHCL on RBC suspension viscoelasticity, cell shape, volume and density were also investigated. Four indices of RBC aggregation, induced by autologous plasma or 3 g% dextran T70, were evaluated by a computerized light transmission method, and the viscous and elastic components of the complex viscosity were determined by oscillatory viscometry. Low concentrations of PRHCL (8 x 10(-5) to 8 x 10(-4) M) significantly (p less than 0.05 or better) reduced the extent of aggregation (maximal decrease of 22% at 8 x 10(-4) M), but did not alter the viscoelastic components, cell shape, volume or density. The anti-aggregating effect of PRHCL (8 x 10(-4) M) in plasma significantly (p less than 0.005) decreased with time; this temporal effect was abolished by addition of eserine (1 x 10(-4) M). High concentrations of PRHCL (8 x 10(-2) M) caused: 1) increased extent of aggregation and decreased strength of the aggregates (p less than 0.01 or better); 2) elevation of both viscoelastic components for cells in plasma or buffer; 3) a discocyte-stomatocyte shape change; 4) decreased cell density (p less than 0.001) without alteration of cell volume. Our results at low concentrations of PRHCL suggest a mechanism based on an increase of RBC negative surface potential; at the highest concentration, the effects are most likely due to altered cell shape and deformability, and to decreased RBC negative surface potential.

Differences in red cell behaviour between patients with Raynaud's phenomenon and systemic sclerosis and patients with Raynaud's disease.

The 'filterability' and electrophoretic mobility of erythrocytes from 42 patients with systemic sclerosis and Raynaud's phenomenon were studied and compared with the findings from 24 patients with Raynaud's disease and 26 normal controls. Red blood cells from patients with systemic sclerosis and Raynaud's phenomenon were less filterable (P less than 0.0001) and had decreased electrophoretic mobility (P less than 0.001) compared with erythrocytes from patients with Raynaud's disease and the controls. There was no significant difference between the values from the patients with Raynaud's disease and the controls. These results indicate that measurement of erythrocyte filterability and electrophoretic mobility may be useful in the differentiation of patients with Raynaud's disease who have no underlying collagen disease from those who have Raynaud's phenomenon in association with systemic sclerosis.

Ex vivo effects of nifedipine, nisoldipine and nitrendipine on filterability of red blood cells from healthy volunteers.

In a balanced, randomised and double-blind trial, the effects of single oral dose of nifedipine, nitrendipine and nisoldipine were compared with placebo in eight healthy volunteers. Red cell filterability, measured with a gravity driven filtration technique, was not significantly altered by any of the three calcium antagonists compared with placebo, when RBCs were filtered within 2 h of venepuncture. Storage of RBCs for 24 h at 25 degrees C, however, significantly reduced RBC filterability compared with 2 h (P less than 0.05), but the reduction after nifedipine and nitrendipine was significantly (P less than 0.05) less than after placebo. The above results demonstrate an effect of calcium antagonists on filterability of stored RBCs.

Plasma or serum from patients with systemic sclerosis alters behaviour of normal erythrocytes.

Incubation of washed normal erythrocytes with fresh or frozen plasma or serum from patients with systemic sclerosis (SS) significantly decreased the filterability of the cells, whereas the incubation of homologous normal serum or plasma with washed erythrocytes did not alter the filterability of these cells. The endothelial cell adherence of normal erythrocytes was increased by 3.5- and 2.6-fold (p less than 0.01) respectively when plasma or serum from patients with SS rather than that from normal controls was added to the incubation medium. Further investigation and isolation of the reactive material(s) present in the serum and plasma of patients with SS, which affect the deformability and endothelial cell adherence of erythrocytes, may be helpful in understanding the pathogenesis of the disease.

Effects of verapamil on calcium-induced rigidity and on filterability of red blood cells from healthy volunteers and patients with progressive systemic sclerosis.

The effects of verapamil on calcium-induced decrease deformability of red blood cells (RBCs) and on the filterability of RBCs from healthy volunteers and patients with progressive systemic sclerosis (PSS) were investigated in vitro using a gravity driven filtration technique. The filterability of RBCs was increased by verapamil 1 microgram/ml (P less than 0.01) in healthy volunteers (P less than 0.05) and in patients with PSS (P less than 0.05). A high concentration of verapamil (200 micrograms/ml) caused an 80% reduction (P less than 0.05) in the filterability of RBCs from healthy volunteers. The filterability of RBCs stored at 4 degrees C for 24 h was increased by 1 microgram/ml verapamil (P less than 0.05). Verapamil (1 microgram/ml) prevented the decrease in deformability of RBCs due to an increase in either extracellular or intracellular calcium concentrations (P less than 0.05). By increasing red cell filterability verapamil may be useful in the treatment of PSS and other peripheral vascular diseases where decreased red cell deformability may play an important role in the pathogenesis.

The effect of pentoxifylline on filterability of normal red blood cells and their adhesiveness to cultured endothelial cells.

The effects of pentoxifylline on filterability of normal red blood cells (RBCs) and their adhesiveness to cultured endothelial cells were investigated. In a balanced randomized and double blind trial, six healthy volunteers received 400 mg pentoxifylline or matching placebo 2 h before blood samples were taken. Filterability of RBCs of the subject while on pentoxifylline was significantly increased at 25 degrees C and 18 degrees C. Lowering of the filteration temperature to 18 degrees C significantly decreased filterability of RBCs. In vitro studies showed that 12 micrograms/ml pentoxifylline significantly increased RBC filterability and also partially prevented the effect of decreasing temperature on RBC filterability. 12 micrograms/ml pentoxifylline significantly decreased the adherence of normal RBCs to cultured endothelial cells. Our results suggest that in addition to increasing filterability of RBCs, pentoxifylline also decreased the adherence of RBCs to endothelial cells and this may contribute to its therapeutic effect.