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T S Kickler

Publications and source records attributed to T S Kickler.

At least 37 records · Page 2Linked to original sources
PubMed1996

The role of HLA antibodies in neonatal thrombocytopenia: a prospective study.

The role of HLA antibodies in neonatal alloimmune thrombocytopenia is controversial. We prospectively studied the sera of obstetric patients at delivery for HLA antibodies and correlated their presence with umbilical cord blood platelet counts. We studied 493 births at The Johns Hopkins Hospital comprising of 357 African American, 115 Caucasian, and 21 babies of other racial groups. One hundred and thirty nine mothers had HLA antibodies. Of these HLA alloimmunized mothers, only ten infants had platelet counts of 150,000/ microL or less. Three hundred and eight mothers with no detectable antibodies gave birth to 27 infants with platelet counts of 150,000/microL or less. Yates corrected Chi square analysis showed no significant relationship between maternal HLA alloimmunization and baby platelet count (p = 0.709). Only 8 of sixty cord sera from babies of HLA alloimmunized mothers were positive for HLA antibodies. The HLA cord blood antibody results were then correlated with the neonatal platelet counts. The Fisher's exact test showed no significant relationship between the presence of HLA antibodies in cord blood samples and neonatal platelet counts (p = 0.232). Although one third (31%) of mothers have HLA antibodies, neonatal thrombocytopenia is rarely associated with this finding. However, HLA antibodies can cross the placenta, and in these unusual cases, may be associated with a higher risk of neonatal thrombocytopenia.

Female↗
PubMed1996

Leukocyte reduction of red cells when transfusing patients with autoimmune hemolytic anemia: a strategy to decrease the incidence of confounding transfusion reactions.

Autoimmune hemolytic anemia (AIHA) presents a difficult challenge to clinicians and blood bankers alike. Autoantibodies in the serum significantly complicate serologic evaluation, and necessitate performing procedures such as adsorptions to eliminate the possibility of underlying alloantibodies. In many instances the blood that is issued may be phenotypically similar but remains crossmatch incompatible, generating a considerable degree of anxiety among the clinical staff who are responsible for transfusing the patient. We report a case of warm autoimmune hemolytic anemia (WAIHA) in which the transfusion of red cells was complicated by a febrile transfusion reaction. Evaluation of the reaction resulted in a significant delay in transfusion therapy. Subsequent administration of leukocyte-poor red cells resulted in uneventful transfusions with a good therapeutic response. Retrospective analysis of the pretransfusion sample demonstrated significant levels of anti-neutrophil antibodies. This case resulted in the establishment of our policy to administer all red cell transfusions to patients with autoantibodies (warm or cold) as leukocyte- poor red cells.

Journal Article↗
PubMed1995

Molecular genotyping of fetal platelet antigens with uncultured amniocytes.

OBJECTIVE: Amino acid substitutions in platelet membrane glycoproteins result in alloantigens implicated in neonatal alloimmune thrombocytopenia. We report the use of the reverse dot blot technique to genotype the five major fetal platelet alloantigens from amniotic fluid cells. STUDY DESIGN: We evaluated a patient with Bakb platelet antibodies who had a previous pregnancy complicated by fetal intracranial hemorrhage. The father was heterozygous Baka/Bakb, giving the pregnancy a 50% risk for platelet incompatibility between mother and fetus. Amniotic fluid was obtained at 16 weeks. Deoxyribonuleic acid was extracted from uncultured amniocytes and amplified with polymerase chain reaction. These products were hybridized to filters containing oligonucleotides specific for each of the 10 different platelet antigen alleles. Reactivity was detected with a chromogenic substrate. RESULTS: The reverse dot blot genotyping of uncultured amniocytes revealed the fetus to be Baka/Baka, thus not at risk for neonatal alloimmune thrombocytopenia. CONCLUSION: Precise knowledge of fetal platelet type by amniocentesis could obviate the need for fetal blood sampling and significantly alter prenatal management of neonatal alloimmune thrombocytopenia.

Adult↗
PubMed1995

Gene frequencies of the five major human platelet antigens in African American, white, and Korean populations.

BACKGROUND: The study of the immunogenetics of the human platelet antigens is important to the improvement of diagnosis and genetic counseling and to the development of screening programs for women at risk of having babies with neonatal alloimmune thrombocytopenia. Description of the immunogenetics of the human platelet antigens in some racial groups has been incomplete. STUDY DESIGN AND METHODS: A reverse dot blot technique employing polymerase chain reaction-amplified genomic DNA was applied in genotyping the five major human platelet antigens in the following populations: 100 African American and 100 white women admitted to the obstetric unit at Johns Hopkins Hospital (Baltimore, MD) and 100 inpatients at Yonsei University (Seoul, Korea). RESULTS: The gene frequencies of HPA-2b (Koa) and HPA-5b (Bra) in African Americans were twice those in whites (African Americans: 0.18 and 0.21, respectively; whites: 0.09 and 0.11, respectively). There is a very low gene frequency of the HPA-1b (PIA2) allele in Koreans (0.005). No significant differences were found in the gene frequencies of the human platelet antigens in whites in this series and those in published European studies. CONCLUSION: These studies indicate a higher potential risk for alloimmunization to HPA-2 (Ko) and HPA-5 (Br) antigens in African Americans than in whites. In addition, the low gene frequency of HPA-1b (PIA2) in African Americans and Koreans suggests that alloimmunization to HPA-1a (PIA1) would be very unusual in these populations. These data may provide the basis for planning neonatal alloimmune thrombocytopenia screening programs in certain ethnic populations.

Antigens, Human Platelet↗
PubMed1995

White cell reduction in platelet concentrates and packed red cells by filtration: a multicenter clinical trial. The Trap Study Group.

BACKGROUND: Most previous studies on white cell (WBC) reduction by filtration have been small-scale studies conducted under tightly controlled laboratory conditions. Their results would be the ideal, rather than what might be expected during routine operation. STUDY DESIGN AND METHODS: To obtain information on routine filtration of blood components, data have been collected from a large-scale, ongoing, multicenter clinical trial designed to determine the effectiveness of WBC reduction in or ultraviolet B radiation of platelet concentrates before transfusion in preventing platelet alloimmunization and platelet transfusion refractoriness. The WBC content of blood components both before and after filtration was determined by automated cell counters and a manual propidium iodide-staining method, respectively. Platelet and red cell losses during filtration were measured. RESULTS: The average platelet losses after filtration were 24 +/- 15 percent and 20 +/- 9 percent for apheresis platelets and pooled platelets, respectively. The frequencies at which filtered platelet concentrates contained high levels of residual WBCs (> 5 x 10(6)) were 7 percent and 5 percent for apheresis platelets and pooled platelets, respectively. Further analysis of the platelet filtration data showed that greater numbers of total initial WBCs in the pooled platelets were associated with increased percentages of filtration failure (> 5 x 10(6) residual WBCs). For packed red cells, the average losses during filtration were 23 +/- 4 percent and 15 +/- 3 percent for CPDA-1 units and Adsol units, respectively. The frequencies at which filtered red cells contained > 5 x 10(6) residual WBCs were 2.7 percent for one type of filter and 0.3 percent for another type of filter. CONCLUSION: There were significant losses of platelets during filtration, which could add to the costs of WBC reduction and lead to possible increases in donor exposures. Filtration failures still occurred, despite careful observation of the standard filtration procedures. The number of total WBCs in pooled platelets before filtration has been identified as an important factor in determining the success of WBC reduction.

Blood Platelets↗
PubMed1995

Studies using immobilized platelet glycoproteins for detection of platelet alloantibodies.

The utility of an enzyme linked immunoassay for the detection of platelet alloantibodies to isolated platelet glycoproteins employing commercially available monoclonal antibodies was studied. The authors established the usefulness of the assay for antibodies to platelet alloantigens (APAA) by testing its sensitivity and specificity under different conditions and using a variety of alloantibodies from patients with immune mediated thrombocytopenia. The authors were able to specifically detect a variety of antibodies to platelet alloantigens on different platelet glycoproteins. These antibodies had specificity for HPA-1a (PIA1), HPA-3 (Bak) and HPA-5 (Br). These results show that the APAA is easily adaptable for routine diagnostic testing in hospital laboratories using commercially available monoclonal antibodies and for the diagnosis of alloimmune mediated thrombocytopenic conditions.

Antibodies, Monoclonal↗
PubMed1995

A monoclonal antibody (SZ21) specific for platelet GPIIIa distinguishes P1A1 from P1A2.

Neonatal alloimmune thrombocytopenia (NATP) and post-transfusion purpura (PTP) are acquired bleeding disorders caused by alloimmune thrombocytopenia. In most cases, the thrombocytopenia is due to an alloantibody directed against the platelet glycoprotein IIb-IIIa (GPIIb-IIIa) complex. During the course of routine studies on the role of GPIIb-IIIa in inherited and acquired bleeding and thrombotic disorders, we unexpectedly identified an individual whose platelets reacted by non-reduced Western blot analysis with anti-GPIIIa polyclonal antisera, but did not react with a commercially available monoclonal antibody (SZ21) specific for GPIIIa. We screened all 14 GPIIIa exons for possible nucleotide changes which might alter amino acids and found variations in only exons 3 and 10. Nucleotide sequencing revealed that only the exon 3 alteration changed the predicted amino acid sequence. This variation was caused by homozygosity for the uncommon P1A2 allele of the GPIIIa gene. Platelets from two additional unrelated normal individuals known to be homozygous for P1A2 also lacked reactivity with SZ21 by Western blot. Using flow cytometry with intact platelets, we observed a markedly reduced binding of SZ21 to platelets with the P1A2 genotype. Scatchard analyses indicated that SZ21 bound to P1A1/A1 platelets with a Kd of approximately 8.26 x 10(-10) M, and to P1A2/A2 platelets with a Kd of approximately 5.58 x 10(-9) M. Thus, we have characterized a readily available monoclonal antibody able to distinguish between the two P1A alleles of the GPIIIa gene. Because incompatibility for this platelet polymorphism is the most common cause of neonatal alloimmune thrombocytopenia and posttransfusion purpura, and because platelet immunophenotyping reagents lack specificity and are not easily available, this monoclonal antibody could facilitate the management of patients with these disorders.

Antibodies, Monoclonal↗
PubMed1995

A quantitative assay for subclassing IgG alloantibodies implicated in hemolytic disease of the newborn.

Traditionally, IgG subclassing has been performed using qualitative assays. Quantitation of IgG subclasses may have prognostic value in evaluating alloimmunized pregnancies. A quantitative enzyme-linked immunosorbent assay (ELISA) was implemented for measuring IgG subclasses of red blood cell (RBC) antibodies (AB) isolated by adsorption/elution from the sera of alloimmunized pregnant women. The assay is a sandwich enzyme immunoassay using monoclonal antibodies specific for the relevant IgG subclasses and anti-human IgG peroxidase conjugate to quantitate the amount of bound IgG. The sensitivities of the assay for IgG1, 2, 3, and 4, respectively, were 4, 23, 4 and 2 micrograms/l. The results for each subclass for a given AB were expressed as a percentage of the total. In a series of pregnant mothers with ABs: E (4), Fya (2), Jka (1) and S (1), the mean percentage +/- 1SD of each subclass was: IgG1 61 +/- 34; IgG2 14 +/- 22; IgG3 18 +/- 28 and IgG4 4 +/- 17. IgG1 or IgG3 accounted for greater than 50% of the AB subclass distribution in 5 cases that resulted in hemolytic disease of the newborn (HDN). Although only a small number of samples was studied, changes in the concentrations of IgG1 or IgG3 during gestation suggest a correlation with the presence or absence of HDN. The ELISA may be used to quantitate the IgG subclasses of RBC ABs and may be valuable in predicting the severity of HDN in alloimmunized pregnancies.

Blood Group Antigens↗
PubMed1994

Expression and purification of functional recombinant epitopes for the platelet antigens, PlA1 and PlA2.

The platelet antigens, PlA1 and PlA2, are responsible for most cases of posttransfusion purpura (PTP) and neonatal alloimmune thrombocytopenia (NAIT) in the caucasian population and are determined by two allelic forms of the platelet glycoprotein GPIIIa gene. To study the interaction between these antigens and their respective antibodies, we inserted the sequence that encodes the signal peptide and the N-terminal 66 amino acids of the PlA1 form of GPIIIa into the expression vector pGEX1. To express the PlA2 antigen, nucleotide 196 of the PlA1 coding sequence was mutated to the PlA2 allelic form. When transformed and induced in Escherichia coli, the two constructs produce glutathione S-transferase (GST)/N-terminal GPIIIa fusion proteins, one containing leucine at position 33 (PlA1), the other proline (PlA2). These proteins are easily purified in milligram quantities using glutathione-Sepharose and react specifically with their respective antibodies by immunoblot and enzyme-linked immunosorbent assay. Antigenicity of the PlA1 fusion protein in reduced glutathione increases with time; moreover, the addition of oxidized glutathione accelerates this process, presumably because of formation of the native disulfide bonds. Neutralization assays indicate that the PlA1 fusion protein competes for all of the anti-PlA1 antibody in the serum of patients with PTP and NAIT that is capable of interacting with the surface of intact platelets. This study shows that the GST/N-terminal GPIIIa fusion proteins contain conformational epitopes that mimic those involved in alloimmunization, and that regions other than the amino terminal 66 amino acids of GPIIIa are not likely to contain or be required for the development of functional PlA1 epitopes. Furthermore, these recombinant proteins can be used for the affinity-purification of clinical anti-PlA1 antibodies and specific antibody identification by western blotting, making them useful in the diagnosis of patients alloimmunized to PlA1 alloantigens.

Alleles↗
PubMed1994

The use of polymerase chain reaction to determine fetal RhD status.

OBJECTIVE: Our purpose was (1) to establish the accuracy of a deoxyribonucleic acid amplification method in determination of RhD status in adult blood samples, including weak D variants (previously referred to as Du) and a D mosaic, and (2) to apply the method to determine fetal RhD status in alloimmunized pregnancies. STUDY DESIGN: Twenty-five adult blood samples, including five weak D variants and one D mosaic, were analyzed with a polymerase chain reaction to determine RhD type. The method was then applied to amniotic fluid samples obtained by amniocentesis from three RhD-negative women with known RhD sensitization. RESULTS: RhD type determined by polymerase chain reaction for all adult blood samples agreed with serologic typing results. All weak D variants and the D mosaic gave results consistent with RhD positivity. Fetal RhD status was determined in each of the three alloimmunized pregnancies, and obstetric management decisions were made on the basis of these results. CONCLUSIONS: This polymerase chain reaction method allows rapid and accurate determinations of fetal RhD status by amniocentesis. Fetal blood sampling or serial amniocenteses may be avoided when the fetus is RhD negative, and plans for surveillance and intervention can be confidently made if the fetus is RhD positive. However, before the widespread use of this assay, its sensitivity and specificity must be established. Because weak D variants and a D mosaic demonstrated RhD-positive status by polymerase chain reaction, the method described is applicable to these RhD variants.

Adult↗
PubMed1994

Variable degrees of suppression of hemoglobin S synthesis in subjects with hemoglobin SS disease on a long-term transfusion regimen.

The objectives of this study were to quantify the amount of blood required to suppress synthesis of hemoglobin S (HbS) in patients with hemoglobin SS on a long-term transfusion regimen and to evaluate factors that might contribute to variations in transfusion-induced patterns of responsiveness. Eleven patients with hemoglobin SS (age range, 2 years 4 months to 19 years 9 months) who had had a cerebrovascular accident were monitored during a period of 1 1/2 to 4 years for HbS percentages, reticulocyte percentages, the amount of erythrocytes infused, and weight. From these data the amount of blood necessary to maintain the HbS concentration at less than 30% was expressed as units of packed erythrocytes administered per week per kilogram of body weight. Percentage of HbS were significantly lower in three subjects than in the other eight (6.1 +/- 0.6 vs 23.0 +/- 2.1; p = 0.0009) as were the reticulocyte percentages (2.9 +/- 0.3 vs 7.9 +/- 0.7; p = 0.0021). However, there were no significant differences between pretransfusion hematocrit (0.278 +/- 0.012 vs 0.281 +/- 0.01; p = 0.90) and units of erythrocytes given per week per kilogram (0.0147 +/- 0.0008 vs 0.0156 +/- 0.0009; p = 0.58). Factors explored to define the reason that HbS synthesis was more easily suppressed in some patients than in others included measurements of serum chemistry values and erythropoietin, identification of erythrocyte alloantibodies, and a survey for Howell-Jolly bodies. No significant differences were seen. Although the reasons for the marked variation in transfusion-induced depression of HbS synthesis are unclear, this study emphasizes the importance of determining the units of packed erythrocytes needed per week per kilogram and correlating this value with the pretransfusion HbS percentage. By doing so, one can select the minimal amount of blood necessary to achieve the desired HbS percentage and thereby decrease the risks of transfusion.

Adolescent↗
PubMed1993

Effect of extracorporeal membrane oxygenation on platelets in newborns.

OBJECTIVES: To examine the effect of extracorporeal membrane oxygenation (ECMO) therapy on platelet function and number and to determine the in vivo survival of transfused platelets during ECMO. DESIGN: Prospective study of all neonates treated with ECMO during a 6-month period. SETTING: Neonatal intensive care unit in a tertiary referral center. PATIENTS: Ten infants, < 1 wk of age, with respiratory failure requiring ECMO. INTERVENTIONS: None. MEASUREMENTS: Platelet counts were measured at timed intervals to determine the effect of ECMO on platelet counts. The change in platelet counts after transfusion was also measured. Whole blood platelet aggregation studies were performed to determine the effect of ECMO on platelet function. MAIN RESULTS: A mean decrease of 26% from the baseline platelet count was found 15 mins after the initiation of ECMO, with an additional mean decrease of 16% by the end of 1 hr (p < .05). Fifteen minutes after platelet transfusions, a 17% mean increase in platelet counts was measured (p < .05). One hour after platelet transfusion, the platelet count had decreased to pretransfusion values. Platelet aggregation studies 15 mins after starting ECMO showed a 46% mean decrease in the response to collagen from baseline (p > .05), and a significantly (p < .05) reduced response to ristocetin and to adenosine 5'-diphosphate. Platelet adenosine triphosphate release was also significantly reduced. Single-unit platelet transfusions failed to correct platelet aggregation abnormalities. Resolution of platelet aggregation abnormalities and normalization of platelet counts occurred 8 hrs off ECMO. CONCLUSIONS: The results of these studies show that qualitative and quantitative platelet changes occur while a neonate is receiving ECMO. The survival rate of transfused platelets is reduced, and the acquired platelet dysfunction is not reversed by the transfusion of platelets with normal function while the patient is receiving ECMO.

Adenosine Triphosphate↗
PubMed1993

Compatibility testing problems associated with bovine thrombin-treated plasma.

Plasma converted to serum by the addition of bovine thrombin prior to compatibility testing agglutinated all donor red cells. This finding prompted an investigation of bovine thrombin- associated incompatibility that showed that (1) thrombin derived from bovine plasma contains IgG antibodies directed against all human red cells, and (2) excess (> 50 units/mL) bovine thrombin used for conversion of plasma to serum may cause hemagglutination and erroneous serologic test results because of the presence of heteroagglutinins.

Journal Article↗
PubMed1993

Efficacy of immune globulin in preventing complications of bone marrow transplantation: a meta-analysis.

To assess the efficacy of immune globulin in preventing CMV infection, interstitial pneumonia, GVHD and death after BMT, we reviewed and synthesized data from 12 published studies (with 1282 patients) in which immune globulin was used prophylactically in BMT patients, controls were included and clinical outcomes were assessed. Data synthesis indicates that immune globulin significantly reduces fatal CMV infection (odds ratio (OR) 0.47, 95% confidence interval (CI) 0.23-0.99), CMV pneumonia (OR 0.61, CI 0.42-0.89), non-CMV interstitial pneumonia (OR 0.57, CI 0.35-0.95) and total mortality (OR 0.74, CI 0.55-0.99). The reduction in acute GVHD was not quite significant (OR CI 0.45-1.02). Complications decrease with both hyperimmune and conventional immune globulin. For CMV-negative transplant recipients, immune globulin decreases symptomatic CMV infection (OR 0.55, CI 0.31-0.94) and interstitial pneumonia (OR 0.34, CI 0.15-0.77). For CMV-positive recipients, immune globulin prevents interstitial pneumonia (OR 0.45, CI 0.26-0.80) but not symptomatic CMV infection (CI 0.41-2.80). We conclude that immune globulin is efficacious in preventing major complications of BMT in both CMV-negative and CMV-positive recipients.

Adult↗
PubMed1992

Chorionic villus sampling for fetal Rh typing: clinical implications.

OBJECTIVE: The prenatal Rh typing of red blood cells obtained by chorionic villus sampling was performed with an immune rosette technique. STUDY DESIGN: Ten Rh-negative pregnant women undergoing chorionic villus sampling at 9 to 11 weeks' gestation were studied at a large referral hospital. RESULTS: Accurate Rh phenotyping may be done on red blood cells obtained from chorionic villi weighing 2 to 8 mg. The preparations were shown to be greater than 90% fetal in origin as demonstrated by radial immunodiffusion quantitation of fetal hemoglobin-containing cells. Of the 10 fetuses typed in the first trimester nine of the pregnancies were carried to term. In all cases typing of red blood cells confirmed the antenatal fetal red cell typing. CONCLUSIONS: This study shows that antenatal Rh phenotyping may be performed as early as 9 to 11 weeks' gestation. This technique may be used in the management of pregnancies complicated by Rh incompatibility.

Blood Grouping and Crossmatching↗
PubMed1992

Elevated platelet-associated IgG in PlA1-negative mothers following sensitization to the PlA1 antigen during pregnancy.

Between 1984 and 1990, we studied 25 infants with neonatal alloimmune thrombocytopenia caused by alloimmunization to the PlA1 alloantigen. We investigated whether mothers of these infants developed elevated platelet-associated immunoglobulin (PAIgG) in addition to anti-PlA1. Eight of the women were found to have PAIgG which persistent at least 7-10 days postdelivery. Eluates prepared from six of the women's platelets reacted with PlA1-positive and PlA1-negative donor platelets, and platelets from donors with Glanzmann's thrombasthenia. None of the women with elevated PAIgG had thrombocytopenia, eclampsia, or infections. Two women were found to have autoreactive antibodies present in plasma. These results indicate that elevated PAIgG may be found in women immunized to the PlA1 antigen. Some women may also have autoreactive antiplatelet antibodies in their plasma. These findings may lead to confusing serologic findings in evaluating the cause of neonatal alloimmune thrombocytopenia.

Antigens, Human Platelet↗
PubMed1992

IgM autoagglutinins in warm autoimmune hemolytic anemia: a poor prognostic feature.

The presence of both complete IgM autoagglutinins and IgG autoantibodies in warm autoimmune hemolytic anemia (AIHA) is an uncommon finding. Over a 6-year period, only 5 of 115 (4.1%) patients with AIHA had IgM and IgG autoantibodies. In 3 of the 5 cases, the complete IgM autoagglutinins reacted up to 30 degrees C and these patients responded well to corticosteroid or other therapies for warm AIHA. The 2 patients who had warm (37 degrees C) reactive IgM autoagglutinins, were refractory to corticosteroids, splenectomy and cytotoxic drugs, and died due to the complications of hemolytic anemia. The data in these 5 cases suggest that the thermal amplitude of the IgM antibody in these unusual AIHA cases may be predictive of refractoriness to therapy and poor clinical outcome.

Adult↗