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Immunoplatelet counting: potential for reducing the use of platelet transfusions through more accurate platelet counting.

Research is required to determine the optimal approach for prophylactic platelet transfusions in patients with haematological malignant disorders. It has been suggested that thresholds for prophylactic platelet transfusions of platelet counts below 10 x 109/l should be investigated, as these may be equivalent in clinical effectiveness and associated with lower costs and fewer complications. An important concern in such investigation is the accurate estimation of platelet counts below 10 x 109/l. This study aimed to further examine the potential reduction in platelet usage that could be made if a lowered platelet transfusion threshold of 5 x 109/l was used in conjunction with an immunoplatelet counting method. Clinical and laboratory data from 130 haematology patients were used. Standard platelet counting was performed using Bayer H3 and ABX Argos analysers. Immunoplatelet counting was performed by flow cytometry using anti-CD61. The potential for reducing platelet transfusions included consideration of clinical criteria that influence prophylactic platelet transfusion use. The results indicated that the use of an immunoplatelet count with a 5 x 109/l platelet transfusion threshold would potentially reduce the number of transfusions by 10.4% in comparison with a 10 x 109/l threshold and standard automated platelet counting with the ABX Argos analyser, and increase the number of transfusions by 5.4% in comparison with the same threshold using the Bayer H3 analyser. The immunoplatelet count may aid the clinical decision to transfuse platelets, but would not necessarily lead to a reduced use of platelet transfusions.

Flow Cytometry↗

Comparison of manual and automated methods for determining platelet counts in dogs with macrothrombocytopenia.

Platelet counts were performed in 43 Cavalier King Charles Spaniels (CKCS, a breed predisposed to macrothrombocytopenia) and in 10 control dogs using 3 automated systems and 3 manual methods (erythrocyte-lysing agents + counting chamber or evaluation of blood smear). Good correlations were found between platelet counts using all methods (all P < 0.0001; R2 = 0.71-0.85). Best correlations were found between the manual methods. Significantly larger platelets were found in CKCS with platelet count < or = 100,000/microl when compared with control dogs and CKCS with platelet count > 100,000/microl (both P < 0.0001). All platelet counts--except when made with the 2 counting chamber methods--were underestimated at platelet counts < or = 100,000/microl.

Animals↗

Determination of the rate of reduction in platelet counts in recipients of hematopoietic stem and progenitor cell transplant: clinical implications for platelet transfusion therapy.

BACKGROUND: To investigate how a delay between pretransfusion platelet count measurement and actual platelet transfusion affects the assessment of platelet transfusion responses, the rate of reduction in platelet counts was determined in 30 patients with relatively uncomplicated thrombocytopenia. STUDY DESIGN AND METHODS: Fifteen adult and 15 pediatric patients admitted for hematopoietic stem and progenitor cell transplantation were studied. Platelet counts before and after myeloablative conditioning and after prophylactic platelet transfusions were determined and studied as a function of time. The rates of reduction in platelet counts were determined by linear regression analysis. RESULTS: Platelet counts were reduced at linear rates after myeloablative conditioning or prophylactic platelet transfusion in all 30 patients. The average rates of reduction in platelet counts after myeloablation were 1261 +/- 583 and 1070 +/- 492 platelets per microL per hour (mean +/- SD) for adult and pediatric patients, respectively. The average rate of reduction after platelet transfusions during the thrombocytopenic phase was 740 +/- 280 and 820 +/- 288 platelets per microL per hour (mean +/- SD) for adult and pediatric patients, respectively. The rates of reduction in platelet counts between the two phases were significantly different in the two age groups (adult, p < 0.0001; pediatric, p < 0.015) and were proportionally correlated with initial platelet counts immediately before myeloablation and after prophylactic platelet transfusions. CONCLUSION: The rate of reduction in platelet count can have a significant impact on the evaluation of platelet transfusion responses when there is a delay between pretransfusion measurement of platelet count and the initiation of platelet transfusion. In addition, the rate of platelet reduction determined from this study can be used to confirm an accelerated rate of platelet consumption in thrombocytopenic patients.

Adolescent↗

Effects of phototherapy on platelet counts in low-birthweight infants and on platelet production and life span in rabbits.

The effects of conventional phototherapy on platelets in vivo were studied in rabbits and in low-birthweight infants. Rabbits were divided into three groups and individually treated continuously for 96 hours, with blue light, daylight, or no phototherapy. Incorporation of selenomethionine Se75 was used to evaluate platelet production and life span. Platelet turnover was significantly increased in the blue-light phototherapy group, and was also increased in the daylight group but to a lesser degree. Daily platelet counts and hematocrits were performed in infants weighing less than 2,000 gm at birth randomized at 24 +/- 12 hours of life to receive daylight phototherapy continuously for 96 hours (31 infants) or no phototherapy (26 infants). In 12 treated infants (38.7%) platelet counts fell below 150,000/cu mm whereas this occurred in only 3 (11.5%) of the controls. These two lower platelet count subgroups differed significantly in mean gestational age and birthweight with the phototherapy group being the more mature. Pre-phototherapy, the mean platelet count of these 12 infants was lower than the mean of the entire control group, suggesting that these infants had borderline marrow reserve before phototherapy was given. These results suggest that phototherapy increases the rate of platelet turnover. When bone marrow compensation is inadequate, the platelet count may fall.

Animals↗

Automated platelet counts: accuracy, precision, and range.

The automation of platelet counting is essential in laboratories performing a large number of procedures with high precision. Maintaining this precision and establishing accuracy require an understanding of the special problems of counting platelets, including: (1) the large dynamic range of the measurement, (2) the variable size and aggregability of platelets, and (3) the specific requirements of a quality-control method. In evaluating these problems, we designed experiments that measured the linearity and precision of two types of platelet counters, light scattering and electronic aperture impedance, and that evaluated the suitability of commercially available reference materials. The results show that the instrumentation is excellent and the reference materials are good; such that, given a well-planned quality-control method as presented here, the automated platelet count gives a rapid result with precision and accuracy.

Calibration↗

Analytic inaccuracy resulting from hematology specimen characteristics. Three cases of clinically misleading artifacts affecting white blood cell and platelet counts.

Sporadically, hematology analyzers generate grossly erroneous results for one or several parameters of the complete blood count (CBC) because of a characteristic or peculiarity of an individual patient specimen. This article presents three cases of relatively less well-known analytic errors of this sort, falsely elevating white blood cell count and platelet count and decreasing total white blood cell count, as determined on a Coulter Aperture Impedence Analyzer. The type of alteration in the CBC parameters and the instrument histogram abnormalities seen in each of these different types of analytic inaccuracy are discussed.

Adult↗

The platelet count in pregnancy.

The platelet count was measured at approximately monthly intervals during the course of 44 normal pregnancies. There was no evidence of any fall in the platelet count during pregnancy. Any significant change in the platelet count in pregnant women is unlikely to be the result of a normal pregnancy.

Analysis of Variance↗

The peripheral platelet count and ADP-induced platelet aggregation in response to metoprolol and propranolol as studied in young healthy male volunteers.

In previous studies, we have demonstrated that the oral administration of one single dose of metoprolol as well as of propranolol causes a significant increase in the venous platelet concentration. In the present study, we investigated the effect of metoprolol, administered twice daily over a period of 1 week, on the venous platelet count in a group of healthy volunteers. The results showed that the drug significantly raised the venous platelet concentration over the whole study period. Further, the effect of 1 week's metoprolol and propranolol medication on ADP-induced platelet aggregation was investigated in another group of healthy subjects; the results were compared to those obtained during 1 week of placebo medication. It was demonstrated that neither of the two beta-blocking drugs affected ADP-induced platelet aggregability; this was true on the first as well as on the last day of study.

Adenosine Diphosphate↗

Suspension of non-viable fungal spores as a standard for platelet counting.

In the search among biological particles for a standard for counting human platelets, a strain of Absidia corymbifera was found to have spores that resembled platelets. After fixing in formalin and autoclaving the spores had a similar mean cell volume to that of human platelets. A suspension of these killed spores was tested in 100 laboratories and gave consistent results as a standard for human platelet counting. The Absidia corymbifera standard can be used in electronic counting methods but not in sedimentation methods, as the spores will be removed by centrifugation.

Blood Cell Count↗

Multivariate QTL linkage analysis suggests a QTL for platelet count on chromosome 19q.

Platelet count is a highly heritable trait with genetic factors responsible for around 80% of the phenotypic variance. We measured platelet count longitudinally in 327 monozygotic and 418 dizygotic twin pairs at 12, 14 and 16 years of age. We also performed a genome-wide linkage scan of these twins and their families in an attempt to localize QTLs that influenced variation in platelet concentrations. Suggestive linkage was observed on chromosome 19q13.13-19q13.31 at 12 (LOD = 2.12, P = 0.0009), 14 (LOD = 2.23, P = 0.0007) and 16 (LOD = 1.01, P = 0.016) years of age and multivariate analysis of counts at all three ages increased the LOD to 2.59 (P = 0.0003). A possible candidate in this region is the gene for glycoprotein VI, a receptor involved in platelet aggregation. Smaller linkage peaks were also seen at 2p, 5p, 5q, 10p and 15q. There was little evidence for linkage to the chromosomal regions containing the genes for thrombopoietin (3q27) and the thrombopoietin receptor (1q34), suggesting that polymorphisms in these genes do not contribute substantially to variation in platelet count between healthy individuals.

Adolescent↗

Influence of aspirin on platelet count and volume in humans.

Platelet volume and total platelet mass are parameters of platelet activity and have become a prognostic factor in coronary heart disease. We have studied the influence of aspirin on platelet count, volume and total platelet mass in vitro and in a randomized double-blind, placebo-controlled study in 20 healthy young male volunteers in vivo. Aspirin had no influence in vitro on either platelet count, volume or mass. Repeated blood sampling during a 7-day treatment with 250 mg aspirin daily showed an increased platelet count (7.3% on day 1, 3.0% on day 2, 6.8% on day 4 and 9.3% on day 7; p < 0.01) and total platelet mass (7.2, 5.0, 8.6 and 11.5% on days 1, 2, 4 and 7, respectively, p < 0.01). Aspirin treatment without repeated blood withdrawal had no effect. These data indicate that aspirin may affect the circulating platelet mass under certain conditions.

Adult↗

The intrapartum platelet count in patients with HELLP (hemolysis, elevated liver enzymes, and low platelets) syndrome: is it predictive of later hemorrhagic complications?

OBJECTIVE: We wished to determine in patients with HELLP syndrome (hemolysis, elevated liver enzymes, and low platelets) whether (1) there is an intrapartum threshold platelet count that is predictive of immediate or delayed hemorrhagic complications and (2) whether prophylactic platelet transfusion at delivery prevents these outcomes. STUDY DESIGN: In this retrospective, descriptive study, the peripartal courses of 132 patients with class 1 (< or = 50,000/microliters platelet nadir) and 160 patients with class 2 (> 50,000 but < or = 100,000/microliters platelet nadir) HELLP syndrome were reviewed with special attention to laboratory data, evidence of hemorrhage, and details of platelet transfusion therapy. RESULTS: A higher incidence of postpartum hemorrhagic complications (p < 0.001) occurred in class 1 versus class 2 HELLP pregnancies. The tendency to have postpartum incisional bleeding after abdominal or vaginal delivery was related to the degree of thrombocytopenia (p = 0.006). The antepartum threshold platelet count most predictive of subsequent postpartum hemorrhagic complications was < or = 40,000/microliters. The prophylactic administration of platelets does not appear to have either significantly decreased the incidence of postpartum hemorrhagic complications or significantly hastened normalization of the postpartum platelet count. CONCLUSIONS: Although bleeding in the gravid patient is related to more factors than platelet count alone, patients with HELLP syndrome in whom an intrapartum platelet count above 40,000/microliters maintained are unlikely to have clinically significant postpartum bleeding. Patients with intrapartum platelet counts < or = 40,000/microliters, however, are at significant risk for postpartum bleeding, but prophylactic platelet transfusion at delivery does not ensure a significantly lower incidence of postpartum hemorrhagic complications.

Adolescent↗

Reduced platelet counts in neonatal respiratory distress syndrome.

Platelet counts were studied in preterm infants with the respiratory distress syndrome (RDS), excluding patients with significant perinatal and postnatal hypoxia. Counts fell to a nadir on day 4 (p less than 0.000). Exploratory analysis indicated that severity of RDS (levels of FiO2 and mean airway pressure), airleak syndrome, grade of intraventricular hemorrhage (IVH) and low platelet count on day 1 were correlated with low platelet count on days 4 and 5. Further analysis by stepwise removal of each variable demonstrated statistically that FiO2 exerted a unique influence on the change in platelet count between day 1 and days 4/5 (p less than 0.002). It is concluded that the severity of RDS is strongly associated with the fall in platelet count in the first few postnatal days. The effect of IVH on platelet count may be secondary to the effect of the RDS.

Age Factors↗

Acquired von Willebrand disease in patients with high platelet counts.

For patients with high platelet counts an inverse relationship has been established between platelet counts and large vWf multimers in plasma. Platelets are currently thought to be primarily involved in the pathogenesis of this decrease in plasma large vWf multimers, presumably by binding large vWf multimers, resulting in their effective removal from the circulation and/or by degrading (platelet-bound) vWf. In patients with myeloproliferative disorders associated with high platelet counts, a decrease in large vWf multimers in plasma may potentially compromise primary hemostasis. Patients with reactive thrombocytosis exhibit a similar reduction in large vWf multimers in plasma, but their clinical course is usually not complicated by bleeding, probably as a consequence of increased circulating vWf levels due to the behavior of vWf as a reactive protein, which compensates for the relatively decreased levels of large vWf multimers in plasma.

Adult↗

Platelet count in deep saturation diving.

Platelet counts were measured in 10 divers during the course of 4 experimental deep dives (450 and 600 m) and different anticoagulants were tested. The use of sodium citrate as an anticoagulant was associated with artifactual thrombocytopenia, whereas ethylenediaminetetraacetic acid proved to be satisfactory. During one of the dives (450 m) the use of fluorocarbon to remove excess dissolved inert gas before decompression of the samples was systematically tested. Mean platelet count decreased from 272,600 +/- 29,400 mm-3 (mean +/- SD) on Day 7 (450 m) to 177,700 +/- 26,400 mm-3 on Day 12 (360 m). Platelet count had recovered to 209,900 +/- 20,700 mm-3 at the time of surfacing. Mean hematocrit (expressed in percent) increased from 44.7 +/- 2.2 predive to 59.4 +/- 2.0 on Day 7 (450 m) to 40.9 +/- 2.8 at the time of surfacing. These changes were statistically significant (P less than 0.05). Platelet counts on samples that had been degassed with fluorocarbon were not different from samples that had been decompressed without degassing.

Citrates↗

Higher levels of interleukin-6 in cystic fluids from patients with malignant versus benign ovarian tumors correlate with decreased hemoglobin levels and increased platelet counts.

BACKGROUND: Recently, high pretreatment platelet counts and low pretreatment hemoglobin levels were found to be negative prognostic factors in patients with ovarian cancer. Interleukin-6 (IL-6) is a multifunctional cytokine with a diversity of functions leading to the induction of C-reactive protein (CRP), increased platelet counts, and low hemoglobin levels. Different epithelial ovarian cancer cell lines are found to produce varying amounts of IL-6. In this study, a possible relationship between IL-6 levels in cystic fluids of benign and malignant ovarian tumors and pretreatment serum CRP, platelet counts, and hemoglobin levels was evaluated. METHODS: A bioassay and enzyme-linked immunosorbent assay (ELISA) were performed to determine the IL-6 levels in cystic fluids and serum from 42 patients with benign and malignant ovarian tumors. RESULTS: The median IL-6 level was higher in cystic fluids of malignant tumors (n = 21) when compared with cystic fluids of benign tumors (n = 21) (P < 0.01 for bioassay and ELISA). Serum IL-6 levels in patients with malignant tumors were not significantly higher compared with IL-6 levels in patients with benign tumors, whereas CRP levels were higher in patients with malignant tumors (P < 0.01). Cystic fluid IL-6 levels were related to serum CRP levels (r = 0.60, P < 0.01 [bioassay]; r = 0.41, P < 0.01 [ELISA]), and were related inversely to hemoglobin levels (r = -0.57, P < 0.01 [bioassay]; r = 0.54, P < 0.01 [ELISA]). CONCLUSIONS: IL-6 levels are higher in cystic fluids of malignant ovarian tumors compared with benign tumors. The relationship of cystic fluid IL-6 levels with CRP, platelet counts, and hemoglobin levels suggests a possible causative role of tumor-derived IL-6 in the appearance of general side effects of ovarian cancer, which recently have been recognized as prognostic factors.

Adenocarcinoma↗

Comparison of platelet counts in simultaneous venous and capillary blood samples using an automated platelet analyser.

Platelet counts (PC) obtained simultaneously from capillary blood (CB) are generally lower than those from venous blood (VB). We quantified this difference in 17 patients with low platelet counts (LPC) and 18 healthy volunteers with normal platelet counts (NPC). The reproducibility of the counts in these 2 groups of subjects was also evaluated. The mean venous platelet count (VPC) and the mean capillary platelet count (CPC) were 67 +/- 30 x 10(3)/ul (+/- SD) and 61 +/- 23 x 10(3)/ul (p = 0.012) in the LPC, and 264 +/- 44 x 10(3)/ul and 234 +/- 45 x 10(3)/ul (p = 0.00016) in the NPC respectively. The mean difference (d) in the PC between VB and CB were 9.4 +/- 13.1 x 10(3)/ul and 19.4 +/- 17.6 x 10(3)/ul in the LPC and NPC respectively. The coefficients of variation (CV) of double counts for VB and CB were 8.1 +/- 8.3% and 9.8 +/- 8.6% for LPC, and 2.3 +/- 1.6% and 2.5 +/- 2.2% for NPC respectively. In conclusion, VPC was frequently (82.9% or 29/35 cases) higher than CPC. In addition, in patients with LPC, the agreement between VPC and CPC was poor and the counts were less reproducible.

Automation↗

Correlation between bleeding times and platelet counts in women with preeclampsia undergoing cesarean section.

Platelet count and bleeding time and the correlation between these two variables in women with preeclampsia who received epidural or general anesthesia for cesarean section were evaluated. The study included 106 women with preeclampsia who were undergoing cesarean section and 94 healthy, term parturients receiving epidural anesthesia for labor analgesia or for cesarean section. Platelet counts were measured using an automated Coulter Counter, and bleeding times were measured using the modified Ivy bleeding time technique. Platelet count was significantly lower and bleeding time significantly prolonged in patients with preeclampsia compared with the control group (P less than 0.0001). In the preeclampsia group, eight patients (7.5%) had thrombocytopenia (platelet count less than 100,000/mm3), whereas in the control group, all women had a normal platelet count (greater than 150,000/mm3). All but one patient with thrombocytopenia had prolonged bleeding time. In addition, 34% of those women with severe preeclampsia and 13% with mild preeclampsia had prolonged bleeding time, although their platelet count was adequate. In the control group, 2% had abnormal bleeding time in the presence of a normal platelet count. There was good correlation between bleeding time and platelet count only when platelet count was lower than 100,000/mm3 (r = -0.76, P less than -0.02).

Adult↗