ABSTRACT
【Objective】 To investigate the transfusion effect of 0.5-dose of apheresis platelet in pediatric patients. 【Methods】 A total of 195 children who underwent 0.5-dose platelet transfusion from August 2021 to June 2022 were enrolled, and the platelet count within 24 hours before and after platelet transfusion were recorded. They were grouped by gender, disease type, blood product transfusion history, platelet antibody results, platelet storage time and weight to analyze the effect of platelet transfusion. 【Results】 Among 195 cases, 77.4% (151/195) were effective and 22.6% (44/195) were ineffective after 0.5-dose of platelet transfusion. Platelet transfusion more than three times has significant impact on the effectiveness of platelet transfusion, with platelet transfusion efficiency decreased from 80.8% to 65.9% (P<0.05), and CCI decreased from 11.56±8.94 to 8.52±8.42 (P<0.05). The transfusion effect of the HLA and HPA antibodies positive group was significantly lower than the negative group, with CCI decreased from 11.39±8.87 to 7.82±8.59 (P=0.05). Linear regression analysis showed that the effect of platelet transfusion decreased with the increasing of platelet storage time (P<0.05), and the effect of platelet transfusion decreased in children weighing less than 20 kg compared with those weighing more than 20 kg, with the effective rate decreased from 84.1% to 63.5% (P<0.05). Different gender, disease type and the number of red blood cell transfusions had no significant effect on platelet transfusion. 【Conclusion】 The 0.5-dose platelet transfusion has good therapeutic effect in children below 20 kg. The results of HLA and HPA antibodies and the number of platelet transfusions greatly influence the effect of platelet transfusion in children, and the transfusion effect decreases with the increase of platelet storage time.
ABSTRACT
【Objective】 To analyze the factors affecting the efficacy of platelet transfusion and the specificity of HLA-Ⅰ antibodies in tumor patients. 【Methods】 Tumer patients applied for platelet transfusion from November 2019 to July 2021 in Sichuan Cancer Hospital province were screened for platelet antibodies. The transfusion efficacy was evaluated and its influencing factors were analyzed, Antibody positive patients were detected for HLA-Ⅰ antibodies. 【Results】 The positive rates of platelet antibody and HLA-Ⅰ antibody, as well as the platelet transfusion refractoriness rate were 35.2%(50/142), 32.4%(46/142) and 16.9%(24/142), respectively. The platelet refractoriness rate of platelet antibody positive patients was higher than that of platelet antibody negative patients [30.0%(15/50) vs 9.8%(9/92)] (χ2=9.428, P<0.05). Multivariate logistic results showed that gender (χ2=12.608, P<0.001, OR=3.800, 95%CI: 1.819-7.940) was an independent risk factor for platelet antibody production, and platelet antibody(χ2=8.648, P<0.05, OR=3.952, 95%CI: 1.581~9.878) was the independent risk factor for transfusion efficacy. The detection rates of strong positive, positive and weak positive HLA-I antibodies were 69.6% (32/46), 80.4% (37/46) and 97.8% (45/46), respectively. The antibodies with high detection rate were anti-B*15∶12, anti-B*57∶03, anti-B*57∶01, anti-B*13∶02, anti-B*49∶01, anti-B*50∶01, anti-A*25∶01, anti-B*15∶01, anti-B*15∶02 and anti-B*44∶02. There were 19 kinds of HLA-Ⅰ antibodies with MFI≥10 000, including anti-A*02∶01, anti-A*02∶03 and anti-A*02∶06. The CCI values were lower in HLA-Ⅰ antibody positive patients(n=46) than in HLA-I antibody negative patients(n=96) by Wilcoxon test (P<0.05). 【Conclusion】 Among tumor patients who received platelet transfusion, gender(female) was the risk factor for platelet antibody production. Platelet antibody positive patients were prone to platelet transfusion refractoriness. HLA-Ⅰ antibodies were the main platelet antibodies, suggesting that HLA-Ⅰ gene matching platelets could improve the efficacy of platelet transfusion.
ABSTRACT
OBJECTIVE@#To investigate the causes of ineffectiveness of platelet transfusion with monoclonal antibody solid phase platelet antibody test (MASPAT) matching in patients with allogeneic hematopoietic stem cell transplantation and explore the strategies of platelet transfusion.@*METHODS@#A case of donor-specific HLA antibodies (DSA) induced by transfusion which ultimately resulted in transplantation failure and ineffective platelet transfusion with MASPAT matching was selected, and the causes of ineffective platelet transfusion and platelet transfusion strategy were retrospectively analyzed.@*RESULTS@#The 32-year-old female patient was diagnosed as acute myeloid leukemia (high risk) in another hospital with the main symptoms of fever and leukopenia, who should be admitted for hematopoietic stem cell transplantation after remission by chemotherapy. In the course of chemotherapy, DSA was generated due to platelet transfusion, and had HLA gene loci incompatible with the donor of the first transplant, leading to the failure of the first transplant. The patient received platelet transfusion for several times before and after transplantation, and the results showed that the effective rate of MASPAT matched platelet transfusion was only 35.3%. Further analysis showed that the reason for the ineffective platelet transfusion was due to the missed detection of antibodies by MASPAT method. During the second hematopoietic stem cell transplantation, the DSA-negative donor was selected, and the matching platelets but ineffective transfusion during the primary transplantation were avoided. Finally, the patient was successfully transplanted and discharged from hospital.@*CONCLUSIONS@#DSA can cause graft failure or render the graft ineffective. For the platelet transfusion of patients with DSA, the platelet transfusion strategy with matching type only using MASPAT method will miss the detection of antibodies, resulting in invalid platelet transfusion.
Subject(s)
Female , Humans , Adult , Platelet Transfusion , Antibodies, Monoclonal , Retrospective Studies , HLA Antigens , Hematopoietic Stem Cell TransplantationABSTRACT
【Objective】 To study the incidence and specificity of platelet antibody in blood donors in Suzhou, analyze the distribution characteristics of platelet antibody in blood donors in this area, and explore the significance of platelet antibody detection in blood donors to reduce the adverse reactions toplatelet transfusion in clinical. 【Methods】 Platelet antibody detection was performed in 2178 blood donors in this area by solid-phase immunosorbent assay. The antibody specificity of the positive samples was analyzed by commercial kit, and the anti-CD36 antibody positive samples were further identified by flow cytometry and gene sequencing. 【Results】 Twelve positive samples were detected by platelet antibody screening, with a positive rate of 0.55%(12/2 178), including 5 males (0.33%, 5/2 178)and 7 females(1.06%, 7/2 178). Among the positive samples, anti-HLA-Ⅰ antibody was identified in 2 cases, anti-CD36 antibody in 1 case, and the antibody specificity was not identified in the other 9 cases. In one case, the positive rate of anti-HLA-Ⅰ antibody PRA was 31.31%(31/ 99), which was mainly specific to anti-B15, anti-B35 and anti-B40. The positive rate of anti-HLA-Ⅰ antibody PRA in the other case was 45.45%(45/ 99), which was mainly specific to anti-A2, anti-A11, anti-A24, anti-A29, anti-A33, anti-A66, anti-B15 and anti-B35. The blood donor with anti-CD36 antibody was type I CD36 deficiency, and 329_330delAC mutation occurred in exon 5. 【Conclusion】 Through antibody screening and specificity identification, the positive rate of platelet antibody in females was significantly higher than that in males(P<0.05). In addition to the common anti-HLA-I antibodies, anti-CD36 antibody was also detected in type I CD36 deficient blood donor. Therefore, the detection of platelet antibodies in blood donors is of certain clinical significance to reduce the adverse reactions to blood transfusion caused by antibodies in platelet products.
ABSTRACT
【Objective】 To compare three kinds of platelet antibody detection methods used to identify alloantibodies in patients with platelet transfusion refractory(PTR). 【Methods】 The 83 samples from PTR patients were analyzed base on three different methods, including solid phase ELISA, Luminex, and capture. The sensitivity, reproducibility, and consistency of different kits were evaluated. 【Results】 A total of 71 (62 positive and 9 negative) out of 83 samples showed consistent results by three methods. The consistency between Luminex and solid phase ELISA was 95.2% (Kapp value=0.829, P<0.05), between solid phase ELISA and capture method was 85.5% (Kapp value=0.512, P<0.05), and between Luminex and capture method was 90.3% (kappa value=0.636, P<0.05). Among the 12 samples with inconsistent results, 3 cases presented positive results by capture method alone and negative by other methods, which had incompatible cross-matching results with 6 random blood donors; 5 cases with HLA antibodies showed negative results by capture method alone and positive by both Luminex and solid phase ELISA; the other 4 cases were positive in both capture and Luminex, but negative in solid phase ELISA. 【Conclusion】 The consistency of three methods was 85.5%, and each has its limitations. The capture method is rapid, economic and registered domestically, which can be used for preliminary screening.Luminex has the optimal diagnostic performance, which can be used for high-throughput and HPA/HLA antibody analysis. The solid phase ELISA is convenient. The combination of them can detect platelet antibodies effectively.
ABSTRACT
【Objective】 To explore the influence of anti-HLA-Ⅰ with different mean fluorescence intensity (MFI) on the efficacy of HLA-A and -B gene matching platelet transfusion, so as to provide scientific data for clinical platelet gene matching transfusion strategy. 【Methods】 A total of 81 PTR patients had applied for HLA-Ⅰgene matched platelets from the platelet gene database established by our laboratory, and 28 (MFI <5 000) of them needed further avoiding of partial donor-specific antibodies and they were enrolled as the research subjects. According to the platelet MFI value of HLA-Ⅰ antibody-targeting antigen, they were divided into negative transfusion group (MFI <500) (group A) and positive transfusion groups (MFI≥500) ; the latter were further divided into group B (500≤MFI <1 000), group C (1 000≤MFI <3 000) and group D (MFI≥3 000) according to MFI value. Corrected count increment (CCI) in platelet count was used to compare the platelet transfusion effect in 4 groups. 【Results】 Among 28 platelet recipients with MFI <5 000, 19(67.86%) patients successfully received 72 effective transfusions. The first CCI (×109/L) in groups A, B, C and D were 10.27±7.46, 7.58±4.75 (P>0.05), 17.36±7.63 (P>0.05) and -0.77±2.30 (P<0.05), respectively. There was no statistical difference among group A, B and C. 【Conclusion】 The application of HLA-Ⅰ gene matching platelets in PTR patients can adjust the MFI threshold(<2 000) appropriately according to the patient′s condition without compromising the platelet transfusion effect.
ABSTRACT
【Objective】 To analyze the status of the platelet antibody screening and crossmatch in Chengdu in 2019, so as to further improve the corresponding detection strategy to improve the clinical transfusion efficacy. 【Methods】 The patients underwent platelet antibody crossmatch in Chengdu Blood Center in 2019 were selected as research objects Platelet antibody screening and crossmatch were performed by solid-phase agglutination technique, and the sample size, the incidence of platelet antibod, age, blood group, seasonal chracteristics, hospital levels, ratio of repeated crossmatch and the transfusion efficacy were analyzed. 【Results】 321 treatment doses of matched platelets after 259 occasions of crossmatch relative to 85 patients were provided. The positive rate of platelet antibody was 87.06%. 64.71% of the patients were over 40 years old, the proportion of ABO group in crossmatch samples was O>A>B>AB, and the crossmatch cases increased each quarter gradually. All samples were provided by tertiary hospitals. 52.94% of the patients needed crossmatch at least twice, and the efficacy rate of matched platelets transfusion was 63.64%. 【Conclusion】 The platelet transfusion efficacy could by improved by platelet antibody screening and crossmatch, so as to avoid the waste of platelets, which deserves active promotion in clinical.
ABSTRACT
【Objective】 To explore platelet antibody production and its influencing factors in common platelet transfusion diseases. 【Methods】 From January 2018 to June 2020, patients who applied for platelet transfusion in the First Affiliated Hospital of Chongqing Medical University were enrolled. The solid phase agglutination method was used to detect the platelet antibodies of the patients. The platelet antibody positive rate of common platelet transfusion diseases and the types of diseases with higher platelet antibody-positive rates were analyzed. 【Results】 The overall positive rate of platelet antibodies in transfusion candidates was 15.0%. The 21~40 years old age patients(21.5%), females(18%) had a significantly higher platelet antibody positive rate than other ages and males(12.1%) (P<0.05). Besides, patients with a history of blood transfusion and pregnancy also had a higher platelet antibody positive rate. Tumors, blood and hematopoietic organ diseases, and digestive system diseases have higher platelet antibody-positive rates, which were 24.0%, 18.3%, and 16.6%, respectively. The platelet antibody positive rate varies significantly in common platelet transfusion diseases. As to transfused patients stratified by diseases, the platelet antibody positive rate of myeloid leukemia(48.7%)was higher than that of other diseases(P<0.05). In comparison, the platelet antibody positive rate of non-transfusion patients with liver failure and miscellaneous diseases(46.7%)was higher than that of others (P<0.05). 【Conclusion】 The positive rate of platelet antibody is somewhat higher in female, 21~40 years old patients. History of blood transfusion is a key risk factor for the production of platelet antibodies.
ABSTRACT
【Objective】 To investigate the relationship between anti-platelet antibodies, therapeutic effect of intravenous immunoglobulin (IVIG) and Treg/Th17 cells imbalance in children with immune thrombocytopenia (ITP). 【Methods】 The changes and correlation of platelet count and Treg/Th17 ratio before and after IVIG treatment in 60 newly diagnosed ITP children with anti-platelet antibodies and 60 children with primary ITP without anti-platelet antibodies were analyzed. 【Results】 1) Compared with the control group, the efficacy of IVIG treatment was better in children with ITP in the case group(CR+ R cases: 50 vs 32) (P<0.01). 2) After IVIG treatment, platelet count(×109/L)(case: 4.5±2.9 vs 327.4±69.5, control: 4.1±3.2 vs 304.7±75.9), Treg cell level(%)(case: 2.15±1.08 vs 5.09±1.37, control: 2.41±0.92 vs 4.98±1.10), Treg/Th17 ratio(case: 1.10±0.19 vs 7.75±1.11, control: 1.27±0.21 vs 4.69±0.81)significantly increased while Th17 cell level(%) significantly decreased in the 2 groups of children(case: 2.07±1.31 vs 1.37±0.92, control: 2.13±1.18 vs 1.48±1.01); compared with the control group, there was no significant change in Treg, Th17 and Treg/Th17 ratio before and after treatment in the case group (P>0.05), but platelet count increased more significantly (P<0.05). 3) There were 3 positive cases in the control group and 12 negative cases in the case group after IVIG treatment, and IVIG treatment probably had no effect on the positive rate of anti-platelet antibodies in children with ITP (P>0.05). 4) The change in platelet count after IVIG treatment was significantly positively correlated with Treg levels (r=0.49 in the case group and r=0.441 in the control group) and negatively correlated with Th17 cell levels (r=-0.390 in the case group and r=-0.364 in the control group). 【Conclusion】 Anti-platelet antibodies can be used as a predictor of the efficacy of IVIG therapy in children with ITP, but they are not associated with changes in the Treg/Th17 ratio.
ABSTRACT
【Objective】 To investigate the positive rate of platelet antibody in pregnant women and analyze the related factors. 【Methods】 A total of 620 pregnant women who underwent antenatal examination from March 2017 to July 2018 were screened for platelet antibodies by solid phase agglutination method. The relationship between platelet antibody positive rate and blood transfusion history, reproductive history and ABO blood group were analyzed, and the profile of disease were alsodiscussed. 【Results】 55 out of 620 pregnant women presented positive platelet antibody, with a positive rate of 8.87%.The platelet antibody positive rate of pregnant women with ahistory of blood transfusion (14.13%) was higher than those never transfused before(6.65%), and the platelet antibody positive rate of pregnant women with a childbirth history (10.46%) was higher than those didn′t bear before(3.52%), showing statistically significant differences (P0.05). Pregnant women with positive platelet antibodies mainly suffered from diseases such as placenta pravia, scarred uterus, placental implantation and thrombocytopenia. 【Conclusion】 Blood transfusion history, reproductive history and disease type have certain effects on the positive rate of platelet antibody in pregnant women.Screening platelet antibody in pregnant women is of great significance to prevente and reduce miscarriage during pregnancy and the occurrence of neonatal alloimmune thrombocytopenic purpura(NAITP).
ABSTRACT
【Objective】 To analyze the testing results of platelet antibody in peripheral blood of 266 pregnant women, so as to explore the correlation of platelet antibody between adverse pregnancy and the number of pregnancies. 【Methods】 A total of 266 pregnant women, admitted to Shaanxi Provincial People′s Hospital, were selected for platelet antibody detection. They were divided into two groups according to history of adverse pregnancy, and the positive rate of platelet antibody between two groups was compared. They were divided into group A (1 pregnancy), group B (2 pregnancies) and group C (≥3 pregnancies) in parallel, and the positive rate of platelet antibody among three groups was compared 【Results】 The yield rate of platelet antibody in groups with and without adverse pregnancy was 31.81% vs 14.86%, showing statistical significance(P<0.05). The yield rate of platelet antibody of Group A, B and C was 9.09%, 21.62% and 23.65%, respectively, with The significant differences(P<0.05). The statistical analysis of inter-group χ2 test demonstrated a linear trend between the number of pregnancies and platelet antibodies yielding(χ2=7.056, P<0.05). 【Conclusion】 Platelet antibody was correlated with adverse pregnancy, and the yield rate of platelet antibody in the group with adverse pregnancy was higher than that in the group without adverse pregnancy. There was a linear trend between platelet antibodies yielding and the number of pregnancies. The yield rate of antibodies gets higher as the number of pregnancies increased. Taking platelet antibody test as a routine test during pregnancy is conducive to the early detection of clinical adverse pregnancy, as well as the early prevention, early detection and early intervention of fetal/neonatal homoimmune thrombocytopenic purpura and other diseases.
ABSTRACT
Objective To investigate the expression of human platelet antibodies(HPA)in children with immune thrombocytopenia(ITP)at different ages and their clinical significance in the treatment of ITP.Methods Two hundred eighty-eight cases of children who were newly diagnosed as primary ITP and detected with HPA from January 2013 to December 2015 in the First Affiliated Hospital of Zhengzhou University were selected.According to the HPA values,they were divided into <1:10 group,1:10 group and >1:10 group.According to their age,they were di-vided into <3-year-old group and ≥3-year-old group.Data were organized and the relationship among platelet antibody,age,gender and short-term efficiency of treatment was analyzed by using SPSS 21.0 statistical analysis soft-ware.Results In this study,there were 288 cases of children,with male to female ratio of about 1.3:1.0.As to the<3-year-old group and ≥ 3-year-old group,this difference of male to female ratio was statistically significant (respectively 1.93:1.00 and 1.02:1.00,χ2=6.629,P<0.05),and the difference of the HPA positive rate was statistically significant(respectively 72.5% and 59.5%,χ2=5.716,P<0.05);the HPA positive rate of boys and girls respectively was 65.9% and 63.7%,so their diffe-rence of the HPA positive rate was not statistically significant (χ2=0.143,P>0.05).Regarding the short-term efficiency,HPA in <1: 10 group,1: 10 group and >1:10 group was respectively 89.1%,89.1%,100.0%.Statistical analysis suggests:the short-term efficiency of <1:10 group and 1:10 group was basically the same(χ2=0.000,P>0.05);In comparison of <1:10 group with >1:10 group(χ2=4.268,P<0.05),and in comparison of 1:10 group with >1:10 group(χ2=4.411,P<0.05),their differences were statistically significant.Conclusions This study suggests that boys are more susceptible to ITP,espe-cially in the <3 age group.The total positive rate of HPA is higher in <3 years old group.The HPA has a certain guiding significance for the diagnosis of ITP.Compared with the other 2 groups,HPA>1:10 group may have higher short-term efficiency in clinical practice.
ABSTRACT
Objective To investigate the expression and clinical significance of platelet autoantibodies in children with persistent/chronic immune thrombocytopenia (pITP/cITP).Methods Total of 34 children diagnosed with pITP/cITP(14 cases and 20 cases,respectively)in the Department of Hematology and Oncology,Shanghai Children's Hospital from December 2013 to August 2014 were enrolled as the study group,including 20 male and 14 female,the median age of 5 years old.The study also included 20 healthy children (the healthy control group) matched with gender and age,and 24 cases of newly diagnosed ITP (newly diagnosed ITP group) serving as the control groups.Platelet-associated immunoglobulin (PAIg) and platelet-specific autoantibodies on surface of platelets were mea-sured by flow cytometry or flow cytometric bead.Results Significant elevation of PAIgA,PAIgM,PAIgD and specific autoantibodies against glucoprotein(GP) Ⅲa,and GP Ⅱb were demonstrated in children with cITP,as well as specific autoantibodies against GP Ⅰ b,GP Ⅲ a,GP Ⅱ b,and granule membrane protein 140 (GMP140) in children with pITP,compared with the healthy control group(P ＜ 0.05);the levels of GPⅨ,GP Ⅲ a,GMP140 in cITP group and GP Ⅱ b in pITP showed significant declination,compared with the newly diagnosed ITP group(P ＜ 0.05);between piTP group and cITP group,autoantibodies GPⅨ,GP Ⅰ b,GP Ⅱ b,and GMP140 in the latter were much lower(P ＜ 0.05).Significant negative relation between PAIgM and platelet count was found in cITP group (P ＜ 0.05).Receiver operating characte-ristic (ROC) curve analysis showed that the area under ROC curve (AUC) of GP Ⅲ a autoantibodies was larger than that of other platelet-autoantibodies in pITP/ciTP diagnosis.Conclusions Platelet autoantibodies play a significant role in pITP/ciTP,especially platelet-specific autoantibodies,which show a declining tendency in the course and may be the main mechanism.The detection of GPⅢa specific autoantibody is more advantageous against other autoantibodies in the diagnosis of piTP/ciTP.
ABSTRACT
Objective To study the platelet antibody screening and crossing match by surface plasmon resonance(SPR) ,and to find a new way for platelet compatibility testing .Methods The corresponding universal platelet antigen was fixed on the SPR chip surface by the amino coupling method .Platelet antibody positive and negative control serum were analysed by SPR micro-ar-ray ,the stability ,sensitivity and specificity of this technique were discussed ,and compared with MAIPA assay .Finally we used the SPR technology to cross match for ten cases of the platelet antibody positive patients before infusion ,and to evaluate the effect of platelet infusion .Results For the SPR technology ,the stability ,sensitivity and specificity of platelet antibody detection were all better ,106 cases of the repeated platelet transfusion samples were tested by SPR assay and MAIPA method ,there was no signifi-cant difference between them(chi-square=0 .333 ,P>0 .05) ,the total consistency was 97 .2% .The 10 cases of platelet antibody positive patients were crossed match before platelet transfusion by SPR technology ,the good results of 8 cases of them were found by the clinical tracking evaluation ,1 h CCI>7 .5 ,24 h CCI>4 .5 .Conclusion SPR technology for screening platelet antibody mat-ches with MAIPA method in basic quality ,but SPR assay is simple ,rapid ,reliable and intuitive ,label free ,which can satisfy the re-quirements for clinical rapid detection of platelet antibody screening and crossing match .
ABSTRACT
Oxaliplatin is a platinum compound used in patients with gastrointestinal malignancies. It is known to evoke a drug-induced immune-mediated thrombocytopenia, which has not been reported in Korea. We describe a 53-year-old man who developed oxaliplatin-induced immune-mediated thrombocytopenia during chemotherapy for colon cancer. Oxaliplatin-dependent IgG platelet antibodies were detected in his serum on flow cytometry. He was treated with immunoglobulin and corticosteroids without any complications. Physicians should consider oxaliplatin-induced immune-mediated thrombocytopenia, when a sudden, isolated thrombocytopenia develops during chemotherapy with oxaliplatin.
Subject(s)
Humans , Middle Aged , Adrenal Cortex Hormones , Antibodies , Blood Platelets , Colonic Neoplasms , Drug Therapy , Flow Cytometry , Immunoglobulin G , Immunoglobulins , Korea , Platinum , ThrombocytopeniaABSTRACT
BACKGROUND: Platelet antigen and antibody tests have been used in platelet immunological disorders, such as neonatal alloimmune thrombocytopenia (NAIT) and post-transfusion purpura (PTP). Mixed passive hemagglutination (MPHA) method has several advantages, including frozen preservation of platelets, ability to differentiate between anti-HLA and platelet-specific antibodies, and quick and easy interpretation without expensive equipment. In this study, we intended to develop the MPHA method using indicator cells of anti-Rh(D) sensitized group O, Rh+ RBCs. METHODS: We made indicator cells sensitized with anti-Rh(D) with various strengths (1:32 to 1:256) and determined the optimal strength. We determined the sensitivity of the MPHA and compared the results using flow cytometry. We observed the changes of the reaction according to the storage time of indicator cells. RESULTS: The optimal sensitization strengths of the indicator cells were 1:192 and 1:256. MPHA showed strong positive results with 1:8,192 diluted positive control, while the detection limit of flow cytometry was 1:128. Until the second week (mean 16 days), the indicator cells showed good results comparable to those of fresh ones. CONCLUSION: We developed the MPHA method using indicator cells of anti-Rh(D) sensitized group O, Rh+ RBCs. We produced the indicator cells in our own laboratory and obtained platelet panels with rare antigen typing using frozen-stored platelets. This technology will be used effectively for detection of platelet antigens and identification of platelet antibodies and also for platelet crossmatching.
Subject(s)
Antibodies , Blood Platelets , Flow Cytometry , Hemagglutination , Limit of Detection , Purpura , Thrombocytopenia, Neonatal AlloimmuneABSTRACT
Objectives To analyze the mechanisms and implications involved in the generation of platelet associated antibodies in cancer patients.Methods HLA IgG antibody from 116 cancer patients who had received repeated platelet transfusion (more than 3 times) was screened by ELISA and the GTI QUICKSCREEN kit. The positive samples from the initial screening were confirmed for HLA and HPA antibodies using the PAKPLUS reagent. Patients with antibodies were observed for symptoms and clinical effectiveness of platelet transfusions. Results A total of 44 cases of HLA IgG antibody were screened out of 116 samples, with an antibody rate of 37.93%. Among those 44 samples, 18 were confirmed to contain anti-HLA-Ⅰantibody, 8 with anti-HPA antibody, 10 with both anti-HPA and anti-HLA antibodies, and 8 were not identified. The occurrence of antibodies was positively correlated to the times of platelet transfusions and correlated to the improvement of clinical symptoms (P
ABSTRACT
BACKGROUND: Platelet antibody test has been used in the diagnosis and management of immunological platelet disorders and platelet crossmatch. Mixed passive hemagglutination (MPHA) test is a cost-effective, reproducible, easy to perform and convenient method. Anti-IgG coated indicator red cells used for MPHA test have not been made in Korea and those cells have been exclusively donated by Dr. Shibata in Japan. This study was designed to produce domestic indicator cells and to determine its acceptability by comparing to the results obtained with Dr. Shibata's indicator cells. We produced homemade indicator cells by coating human RhD-positive O RBC with human IgG anti-D (check cell) and then coated with rabbit anti-human IgG. METHODS: Sixty three sera from healthy male donors, 58 sera tested positive in platelet suspension immunofluorescence test (PSIFT), and 61 sera tested negative in PSIFT were evaluated by MPHA employing both homemade and Dr. Shibata's indicator cells. RESULTS: The concordance rate between PSIFT and homemade MPHA was 74%. Test results of MPHA with homemade indicator cell showed excellent correlation with Shibata's indicator cell (P=0.002). All of 63 sera from healthy volunteer male blood donors without history of transfusion were tested negative with homemade MPHA method. Test results of MPHA employing homemade indicator cells showed excellent correlation with those of PSIFT (P=2.67x10-8). CONCLUSIONS: Homemade indicator cells we developed in this study were able to replace Dr. Shibata's indicator cells for the MPHA test.
Subject(s)
Humans , Male , Blood Donors , Blood Platelets , Diagnosis , Fluorescent Antibody Technique , Healthy Volunteers , Hemagglutination , Immunoglobulin G , Japan , Korea , Tissue DonorsABSTRACT
Hemodialysis requires anticoagulants to prevent fibrin deposition and thrombus formation in the extracorporeal circuit. Unfractionated heparin (UFH) has been used as a conventional anticoagulant for a long time. But recently, many side effects of heparin have been documented: hemorrhage, thrombocytopenia with or without thrombosis, osteoporosis, skin necrosis, alopecia, and hypersensitivity reactions. In the past decade, low molecular weight heparins (LMWH) have been developed. Compared with UFH, these compounds have a longer plasma half life, less variability in the anticoagulant response to fixed doses, and a more favorable antithrombotic to hemorrhagic ratio. Thus, rationales for using LMWH as an alternative to UFH would be a reduced risk of bleeding complications and simplified routines for heparinization due to a longer half-life of the anticoagulant activity. To evaluate the dfficacy and safety of LMWH as an anticoagulant in hemodialysis treatment, we conducted a prospective crossover study with paired comparison of two different heparins in 18 end-stage renal disease patients undergoing hemodialysis. During the first two months of observation, patients received a single bolus of LMWH (Fragmin(R)) 2,552+/-221 aXa IU/one dialysis session. Then patients were switched to UFH dose regimen comprised of a saline prime, no initial bolus and a continuous infusion of 3,174+/-420 IU/one dialysis session for further two months. All hemodialysis sessions were completed uneventfully. The coagulation values of an anti-factor Xa-specific clotting method (Heptest(R)) from citrated whole blood samples taken 15 minutes after starting hemodialysis were 0.47+/-0.21 U/ml with LMWH and 0.12+/-0.03 U/ml with UFH (p). The prolongation of the Heptest clotting times with LMWH and UFH was 2.86 for LMWH and 2.55 for UFH using the shole blood assay. The mean frequency of clot deposition in dialyzer was similar (1.1 vs 0.87) as well as mean venous compression time at the end of dialysis (5.96 vs 6.23 minutes). The hematologic and biochemical parameters such as hemoglobin, platelet count, triglyceride level, total cholesterol and HDL-cholesterol level did not show any differences between the two heparins. We conclude that a single dose of LMWH is effective and safe in repeated use for hemodialysis and prevents clot formation to a similar degree as UFH.
Subject(s)
Humans , Alopecia , Anticoagulants , Cholesterol , Cross-Over Studies , Dialysis , Fibrin , Half-Life , Hemorrhage , Heparin , Heparin, Low-Molecular-Weight , Hypersensitivity , Kidney Failure, Chronic , Matched-Pair Analysis , Necrosis , Osteoporosis , Plasma , Platelet Count , Prospective Studies , Renal Dialysis , Skin , Thrombocytopenia , Thrombosis , TriglyceridesABSTRACT
In thirty patients with acute leukemia and severe aplastic anemia receiving random single donor platelet transfusions, the development of refractoriness by consecutive platelet transfusions with cytapheresis and its relationship to the appearance of anti-platelet antibodies were investigated. The median number of platelet transfusions inducing refractoriness was 13 times, and 20% of the patients remained unrefractory despite of the repeated multiple platelet transfusions up to 20 to 25 times. The results of anti-platelet antibody tasts by the enzyme-linked immunosorbent assay(ELISA) and immunofluorescent techniques(IFT) showed no statistically significant relationship with the refractoriness (p greater than 0.1). Although there was significant correlation between the results of ELISA and IFT, both tests were insufficient to find out refractoriness even with the use of pooled platelets from multiple donors as target cells. This study shows that 13 single donor platelet transfusions result in refractoriness, that both ELISA and IFT are insufficient to detect refractoriness despite of their significant correlation, and that other methods than these are needed in order to detect alloimmunization.