62例血小板输注无效患者的交叉配型及HLA抗体分析

目的对广州地区血小板输注无效患者进行两种不同试验方法的交叉配型并检测人类白细胞抗原（HLA）抗体特异性。方法对62名临床不同疾病类型血小板输注〉2次的患者分别做流式细胞血小板交叉配型（Flow—XM）试验和经典补体依赖微量淋巴细胞毒交叉配型（NIH．CDC）试验,并进行群体反应性抗体（PRA）分析。结果62例血小板输注无效患者通过NIH．CDC试验,得出血小板配型阳性标本22例,阳性率为35．48％;Flow-XM试验比NIH—CDC试验阳性检出率高,为70．97％,两种配型结果的阳性率比较差异有统计学意义（P〈0．01）。分析配型阳性患者的HLA抗体特异性,除2例为阴性外,其余都证实存在针对供者的HLA抗体。结论Flow—XM试验对于检测HLA抗体具有较高的灵敏度,值得推荐。     

免疫性血小板输注无效患者交叉配合输注的研究

目的 通过比较不同血小板输注策略的输注疗效,探讨血小板抗体筛查及交叉配型对于免疫性血小板输注无效(platelet transfusion refractoriness, PTR)患者的应用价值。方法 采用固相凝集法对临床送检血小板抗体筛查的患者进行血小板抗体检测,提取患者的临床资料如性别、年龄、疾病种类、输血记录等,计算血小板抗体筛查阳性率并分析其分布特点。以血小板输注后增量(the post-transfusion platelet increment, PPI)和血小板校正后增值(the corrected count increment, CCI)为衡量指标,分析比较血小板抗体阳性以及不同输血策略(随机输注、交叉相合、交叉不相合)对患者血小板输注疗效的影响。结果 血小板抗体筛查阳性患者61例(23.55%),且多为血液系统疾病患者;其中,抗体筛查阳性组的PPI(U=18 851,P=0.0720)、CCI(U=14 585,P=0.0183)、血小板输注有效率(χ²=5.691,P=0.017)均低于抗体筛查阴性组。61例抗体阳性患者先后共输注122次血小...     

38840例住院患者血小板抗体筛查分析

目的 通过检测分析血小板抗体在住院患者中的分布特征,探究血小板抗体产生原因,为提高输血质量提供数据支持.方法 选取38840例患者,采用Capture-P固相检测体系检测血小板相关抗体,统计分析抗体阳性率.结果 38840例住院患者中血小板抗体阳性者3989例,阳性率为10.27％.男女性患者阳性率分别为8.7％和11...     

血小板抗体检测的方法比较

随着临床输血事业的发展,血小板(PLT)的使用日益广泛.但是,随着血小板的大量使用,血小板输注无效(refractoriness to platelet transfusion,RPT)已成为困扰临床医生的常见问题.目前对血小板抗体检测的普及程度还很低,为此本文对现有的血小板抗体检测方法进行比较分析,以便为广泛开展血小板抗体检测及血小板配型提供有价值的参数.     

血小板输注无效的影响因素及其预防对策

近年来,随着临床输血技术的不断提高,机采血小板输注大幅增加。然而,在同种免疫及某些非免疫因素作用下,会发生血小板输注无效(platelet transfusion refractorinessPTR),这常常困扰着临床输血工作者。本文就本院2011年1～6月半年内,凡可查询到的血小板输注前后外周血血小板值变化的183位患者输注445袋机采血小板后发生PTR的情况进行了初步分析。     

血液病患儿血小板抗体筛查及交叉配型输注效果分析

目的:分析血液病患儿血小板抗体筛查及交叉配型输注效果。方法:回顾性分析2016-01—2018-05在我院接受多次血小板输注治疗的血液病患儿78例,依据血小板抗体检测结果分为阳性组(n=28)和阴性组(n=50),阴性组直接输注ABO同型血小板,阳性组进行交叉配型,根据配型结果分为未配型组(n=13)和配型组(n=15),未配型组输注ABO同型血小板,配型组输注相同的血小板。所有患儿于输注血小板1h、24h后检测血小板计数,计算血小板增值指数(CCI),分析患者血小板抗体筛查结果、交叉配型及其血小板输注无效(PTR)和非溶血性输血反应(NHTR)情况。结果:阳性组患儿中HLA-I抗体阳性23例(82.14%),HLA-I+HPA抗体阳性4例(14.29%),HPA抗体阳性1例(3.57%)。其中配型组中有HLA-I抗体阳性者12例(80.00%),HLA-I+HPA抗体阳性2例(13.33%),HPA抗体阳性1例(6.67%);而未配型组中有HLA-I抗体阳性者11例(84.61%),HLA-I+HPA抗体阳性2例(15.39%)。阳性组患儿血小板输注后1h、24h的CCI值均低于阴性组(P<0.05),且阳性组PTR、NHTR发生率均明显高于阴性组患儿(P<0.05)。配型组血小板输注后1h、24h的CCI均高于同时点的未配型组,PTR和NHTR发生率均低于未配型组(P<0.05)。配型组血小板输注有效率也明显高于未配型组(P<0.01)。结论:多次血小板输注的血液病患儿,依据血小板交叉配型试验结果选择交叉配型相合的血小板进行输注,可提高血小板的输注效果。     

132例血液病患者血小板输注效果的影响因素分析

目的 探讨影响血液病患者血小板输注疗效的相关因素.方法 对2014年1月至6月在我院输注血小板的132例血液病患者的临床资料进行回顾性分析.以输血后血小板增高指数(CCI值)来评价血小板输注效果,同时对影响血小板抗体阳性率的相关因素如病因、患者性别、输注次数、血型及年龄和影响输注效果的相关因素如抗体强度、血小板种类、保存时间、患者体重及并发症等进行统计学分析.结果 132例血液病患者,抗体总阳性率18.94％,输注1376次,有效率为59.23％;有效组抗体阳性率为8.33％,远低于无效组抗体阳性率31.67％(P＜0.05).不同疾病、输注次数和不同年龄阶段血小板抗体阳性率差异均有统计学意义(P＜0.05).ITP组抗体阳性率最高,随着输注次数的增多,血小板抗体阳性率增高,在三个年龄段中21～40岁年龄段的抗体阳性率最高.血小板抗体的产生与患者性别和血型无关.随着抗体强度和患者体重的增加,血小板输注有效率均降低;辐照机采血小板的输注有效率最高;在同种类型的血液病患者中,无并发症患者的血小板输注疗效明显优于有并发症的患者;血小板保存天数与输注有效率无明显相关性.结论 血液病患者的疾病、年龄、血小板输注次数、血小板抗体的产生、抗体强度、血液成分、体重和并发症对血小板输注均有一定的影响.     

联合分析血小板特异性抗体和瞬逝生物传感器技术检测血小板抗体的方法比较

目的 应用联合分析血小板特异性抗体(specificp platelet antibodies,SASPA)和瞬逝生物传感器技术(evanescent biosensor technology,EVA)分别检测血小板特异性同种抗体和自身抗体,对两者进行方法学比较.方法 收集94例存在HPA-1a、HPA-5b、HLA-I和自身抗体的患者血浆标本,制备血小板膜糖蛋白-血小板特异性抗体复合物,分别用SASPA和EVA方法对复合物进行检测,对SASPA检测所得平均荧光强度值(MFI)和EVA检测所得斜率值(Slope)作统计学分析和比较.结果 SASPA方法对HPA-1a、HPA-5b、HLA-I和自身抗体检测的敏感度分别为74.1％、86.9％、86.9％和40.9％,EVA方法的敏感度分别为88.9％、78.3％、60.9％和72.7％.人类血小板抗原(HPA)-1a抗体检测中,MFI与Slope表现为强相关性,相关性系数为r=O.95(P ＜0.05).结论 SASPA和EVA技术对传统的单克隆抗体特异性血小板抗原固定实验(MAIPA)作了继承和改良,SASPA方法可以用极少量的血小板同时检测多种血小板特异性抗体,提高了检测效率;EVA方法将血小板裂解后的实验时间缩短为10 min,极大地简化了操作过程,减少了检测时间.SASPA和EVA检测方法的建立对临床血小板免疫相关疾病的诊断和血小板输血领域的发展具有深远意义.     

免疫性血小板减少症儿童外周血miRNA-150及血小板膜糖蛋白特异性抗体的表达及临床意义

目的分析免疫性血小板减少症(imumune thrombocytopenia, ITP)儿童外周血中miRNA-150以及血小板膜糖蛋白(glycoprotein, Gp)特异性抗体表达情况和临床意义。方法选择2020年1月至2022年12月济宁医学院附属医院儿科收治的ITP患儿总共98例设为观察组, 另选择同时间济宁医学院附属医院儿科健康体检儿童100例设为对照组, 采用实时荧光定量反应检测并比较两组研究对象外周血miRNA-150以及Gp特异性抗体水平, 分析观察组不同疾病分期和不同疗效患儿外周血中miRNA-150水平及有关抗体表达情况。结果观察组患儿外周血miRNA-150水平高于对照组[(1.14±0.30)比(0.39±0.05), t=24.65, P<0.05]。观察组中新发ITP组的miRNA-150水平、抗GPIIb/Ⅲa抗体阳性率高于持续性ITP组以及慢性ITP组[(1.24±0.22)比(1.15±0.20)、(0.88±0.16), 75.00%比54.05%、61.90%, F/χ2值为14.35、11.38, P值均<0.05]。观察组中治疗后...     

血小板相关抗体检测的应用价值

目的：探讨血小板相关抗体（PAIgG）检测的应用价值。方法：采用流式细胞术分别对42例特发性血小板减少性紫癜（ITP）患者、39例再生障碍性贫血患者及21例其他自身免疫性疾病患者的PAIgG进行检测,并分析其与血小板输注无效率的关系。结果：ITP患者组、再生障碍性贫血患者组及其他自身免疫性疾病患者组PAIgG阳性血小板百分率均明显高于正常对照组（P〈0.01）;PAIgG阳性的ITP患者及再生障碍性贫血患者中的血小板输注无效率均明显高于PAIgG阴性的患者（P〈0.01,P〈0.05）。结论：PAIgG用于ITP诊断敏感性高,特异性差,PAIgG的存在与血小板输注无效存在密切关系,是引起血小板输注无效的重要原因之一。     

Development of de novo HLA donor specific antibodies (HLA-DSA), HLA antibodies (HLA-Ab) and allograft rejection post blood transfusion in kidney transplant recipients

BackgroundBlood transfusion during the post-operative period of kidney transplantation is common as part of a life-saving procedure, especially in the event of acute blood loss. However, there have been conflicting opinions since the pre-cyclosporine era. The risk of sensitization post-transfusion remains the main limiting factor following transfusion in kidney transplant recipients. Thus, the objective of this study is to assess the development of de novo HLA-DSA, HLA-Ab and allograft rejection post blood transfusion.MethodologyThis is a retrospective cohort study recruiting all kidney transplant recipients in South Australia from January 2010 till December 2018. Following that, the incidence of blood transfusion within one week post-operatively were traced (transfusion group). The outcomes were compared with all other transplant recipients (non-transfusion group). Recipient’s demographic, donor characteristics and immunological risk profiles were obtained from the transplant unit database, while the biopsy report, history of blood transfusion, latest serum creatinine and follow-up status was gathered from the electronic medical system (OASIS). The HLA-DSA and HLA-Ab results were collected from the NOMS database. Finally, the survival data were merged with the Australia and New Zealand Dialysis and Transplant (ANZDATA) Registry for South Australia recipients graft survival.ResultsA total of 699 patients were eligible for analysis. The mean age was 50.64 ± 13.23 years old. There were more elderly (>65 years old) and females who needed transfusion. The majority had glomerulonephritis as the primary disease. There was no statistical difference in donor characteristics, cold ischemic time and immunological risk between the transfusion and non-transfusion group. There was no difference in the development of de novo HLA-DSA, HLA-Ab and rejection episodes between the group and the results were consistent in a model adjusted for all potential confounders. Median graft survival in days between the transfusion vs non-transfusion group was 1845 IQR (961,2430) and 1250 IQR (672,2013).ConclusionBlood transfusion under strong immunosuppressive cover within a one-week post-operative period is safe with no significant association with the development of de novo HLA-DSA, HLA-Ab or clinical rejection.     

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/μL or less. Three hundred and eight mothers with no detectable antibodies gave birth to 27 infants with platelet counts of 150,000/μL 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.     

Anti-IgG Antibodies and Antinuclear Antibodies in Allergic Patients

With an indirect immunofluorescencc technique 77 % of 96 patients with type I allergy and 40% of 20 patients with intrinsic bronchial asthma showed positive reactions for IgG ami-IgG antibodies in serum. They were present partly in an aggregated state not directly detectable before treatment with dithiothreitol. The aggregates could be removed by precipitation with polyethylene glycol. The IgG ami-IgG in hyposensitized patients were directed against both F(ab')² and Fe fragments of rabbit IgG. Thirty of the type I allergic patients were examined once during hyposensitization as well. Before treatment 87% and IgG anti-IgG (titres 9–72). After ≥ 13 months of treatment 100% were positive (titres 35–288), Fight patterns were also examined after hyposensitzation had been discontinued for at least 12 months. The titres of IgG ami-IgG had then reverted lo the levels obtained before hyposensitization. Of 116 controls matched for sex and age, 79r had IgG ami-IgG anybodies, is suggested that the production of IgG anti-IgG may be stimulated by the presence of immune complexes and that purity, amount and/or combination of allergens administered during hyposensitization may influence the production of anti-JgG antibodies. Neither IgK anti-IgG nor antinuclear antibodies seem to be of particular significance in allergic patients.     

Deciphering allogeneic antibody response against native and denatured HLA epitopes in organ transplantation

Anti‐HLA donor‐specific antibodies are deleterious for organ transplant survival. Class I HLA donor‐specific antibodies are identified by using the Luminex single antigen beads (LSAB) assay, which also detects anti‐denatured HLA antibodies (anti‐dHLAs). Anti‐dHLAs are thought to be unable to recognize native HLA (nHLA) on the cell surface and therefore to be clinically irrelevant. Acid denaturation of nHLA on LSAB allows anti‐dHLAs to be discriminated from anti‐nHLAs. We previously defined a threshold for the ratio between mean fluorescence intensity against acid‐treated (D for denaturation) and nontreated (N) LSAB, D ≥ 1.2 N identifying the anti‐dHLAs. However, some anti‐dHLAs remained able to bind nHLA on lymphocytes in flow cytometry crossmatches, and some anti‐nHLAs conserved significant reactivity toward acid‐treated LSAB. After depleting serum anti‐nHLA reactivity with HLA‐typed cells, we analyzed the residual LSAB reactivity toward nontreated and acid‐treated LSABs, and then evaluated the ability of antibodies to recognize nHLA alleles individually. We observed that sera can contain mixtures of anti‐nHLAs and anti‐dHLAs, or anti‐nHLAs recognizing acid‐resistant epitopes, all possibly targeting the same allele(s). Therefore, the anti‐HLA antibody response can be highly complex and subtle, as is the accurate identification of pathogenic anti‐HLA antibodies in human serum.     

Anti-centromere antibodies (ACA) in systemic sclerosis patients and their relatives: a serological and HLA study

Autoantibody reactivity to centromere proteins CENP-A, CENP-B and CENP-C was examined in 58 patients with systemic sclerosis (SSc). 218 first degree relatives and 22 spouses, HLA class II typing for HLA-DRB1 and HLA-DQA1 was performed by restriction fragment length polymorphism (RFLP) analysis in 50 families, and HLA-DRB1, HLA-DQA1 and HLA-DQB1 typing was performed by olignucleolitde typing in 44 families. Eleven probands and two relatives had ACA. The two relatives with ACA also had SSc. One relative was an identical twin sister of a pro band with ACA and the other relative was a sister of a proband with ACA. All ACA-positive probands and relatives were female, and all recognized CENP-A, CENP-B and CENP-C. The presence of at least one HLA-DQB1 allele not coding for leueine at position 26 of the first domain appeared necessary, although not sufficient for the generation of ACA, Therefore within SSc families ACA is strongly associated with female gender and disease phenotype, and is at least in part genetically determined.     

Use of Monoclonal Antibodies for the Detection of HLA and DR Antigens on Spermatozoa of Different Species

ABSTRACT: There are conflicting reports about the presence of HLA and DR antigens on human sperm. The difference may be due to the complexity of anti-sera used by earlier researchers. These limitations have been overcome by using monoclonal antibodies in the present studies. The microcytotoxicity test was used to determine the presence of HLA and DR antigens on sperm of different species. It was observed that about 60% of human spermatozoa express HLA antigens and a slightly lower percentage (48%) express DR antigens. These antigens are highly cross-reactive with that of monkey and, to a lesser extent, with that of buffalo. Histocompatibility antigens expressed on epididymal spermatozoa of rat and mouse are also weakly reactive with monoclonal antibodies directed to human HLA and DR.     

Specific Antiphospholipid Antibodies as a Predictive Variable in Patients With Recurrent Pregnancy Loss

PROBLEM: Antiphospholipid antibodies (APLs) consist of very heterogenous autoantibodies. It has not been fully explored what kind of specificities are most relevant to recurrent pregnancy loss. Thus, we investigated the effects of specific APLs on recurrent aborters. METHOD: IgG and IgM antibodies against PE (treated with 1% acetic acid) and five negatively-charged phospholipids were measured by ELISA among 334 recurrent aborters without autoimmune disease. The relationships between APL specificities and subsequent pregnancy outcome were prospectively investigated in 38 recurrent aborters with positive APL who did not receive treatment with prednisolone and aspirin. Antibody levels exceeding the 99th percentile of 280 healthy women were considered positive. RESULTS: Positive IgG and/or IgM APLs were detected in 14%, IgG APLs in 12%, and IgG antibodies against PA, PG, PI, PS, CL and PE, respectively, in 9%, 7%, 7%, 7%, 8%, and 8%. In a prospective study of the 38 untreated patients, fetal loss recurred in 82% of the 33 IgG APL-positive patients, but in 40% of the five patients positive for only IgM APLs. The incidence of fetal loss in the next pregnancy of patients with IgG specific APL-positive against PE, PI, PS, or CI was even higher at 90% and over, and fetal loss recurred in all of 21 patients with two or more IgG APL-positive against PE, PI, PS, or CL. CONCLUSION: These results suggest the possibility that two or more IgG APL-positive value against treated PE, PI, PS, or CL, may be more accurate as a predictive variable than that of only one IgG APL-positive in patients with recurrent pregnancy loss.