抗-CD47单抗对输血相容性检测的干扰及处理

目的 评估抗-CD47单克隆抗体对输血相容性检测的干扰，研究去干扰方法，并对受试者输注效果进行评价。方法 收集本院参与接受天境和信达公司生产的抗-CD47单克隆抗体药物治疗的8名临床试验受试者标本，使用缺乏IgG4的抗人球蛋白试剂Gamma-clone、ZZAP试剂和药物独特型中和试剂等进行血型检测、意外抗体筛查、直接抗人球蛋白试验和交叉配血试验。结果 使用抗-CD47单抗治疗后，8名受试者中有5名ABO血型检定受到干扰；所有受试者直接抗人球蛋白凝集强度呈现2+～4+,且所有受试者意外抗体筛查结果都呈3+～4+。采用缺乏IgG4的抗人球蛋白试剂Gamma-clone的试管法进行意外抗体筛查试验，结果均呈现阴性，交叉配血试验均相合。采用药物独特型中和抗体进行意外抗体筛选，试验结果显示均为阴性，交叉配血试验均相合。使用CD47单抗药物导致贫血的患者均输注2U悬浮红细胞，评价经抗干扰处理后配血相合患者的输血效果，结果显示患者输血有效。结论 受试者使用CD47单抗药物后会干扰输血相容性检测，使用缺乏IgG4的抗人球蛋白试剂和药物独特型中和抗体可去除抗-CD47干扰，受试者输血效果良好。     

抗-CD47单克隆抗体干扰输血相容性检测1例

目的 探讨抗-CD47单抗对输血相容性检测的影响及处理措施。方法 对1名具有抗-CD47单抗用药史的患者血液标本进行输血相容性检测，分析抗-CD47单抗效价，鉴定巯基试剂对抗-CD47单抗的洗脱效果，利用中和抗体及抗球蛋白筛选与患者相配合的血液成分。结果 患者血型鉴定为A型，RhD(+),不规则抗体筛查、DAT试验、交叉配血均为3+或4+。利用中和抗体或Gamma-clone抗球蛋白均可为患者筛选到主侧配血相合的红细胞。患者输注后无不良反应，贫血状况得到明显改善。结论 利用中和抗体或Gamma-clone抗球蛋白处理患者血液标本，可以消除抗-CD47单抗对输血相容性检测的影响，保障临床用血的安全性、及时性和有效性。     

抗-CD38单克隆抗体对输血前检测试验的干扰及其处理措施

目的探讨抗-CD38单克隆抗体对输血前检测实验的干扰及其处理措施。方法收集2名接受抗-CD38单克隆抗体治疗的多发性骨髓瘤患者血样,分别进行ABO和Rh血型抗原定型、直接抗人球蛋白实验、不规则抗体筛选和鉴定、交叉配血实验;将不规则抗体筛选用试剂红细胞、抗体鉴定谱细胞、交叉配血用的献血者红细胞、平行对照用的O型K(+)E(+)红细胞与0.2 mol/L的DTT按照红细胞:DTT为1∶4的比例进行混合后,放入37℃水浴箱水浴30 min;再用上述DTT处理后的红细胞分别对患者血浆进行不规则抗体筛选实验、抗体鉴定实验和交叉配血实验。结果 2名患者的ABO和Rh血型均为O,CCDee;直接抗人球蛋白实验分别为阴性和阳性;未经DTT处理的抗体筛选红细胞、抗体鉴定细胞和献血者红细胞与患者血浆反应均呈阳性;DTT处理后的抗体筛选红细胞、抗体鉴定细胞和献血者红细胞与患者血浆反应均呈阴性;平行对照用的O型K(+)E(+)红细胞经DTT处理后转变为O型K(-)E(+)。结论抗-CD38单克隆抗体治疗多发性骨髓瘤,可干扰患者的血型血清学实验,用0.2 mol/L的DTT灭活红细胞表面的CD38抗原后,可以较好的去除这种干扰。     

抗-cE及抗-Jk^b和抗-Mur混合抗体致疑难配血分析——附1例报告

目的探讨1例疑难交叉配血不合的原因。方法通过ABO血型鉴定、Rh分型、直接抗人球蛋白试验、不规则抗体筛查、抗体特异性鉴定及抗体效价测定对患者配血不合进行分析。结果对照谱细胞反应格局,患者血清中检出抗-cE、抗-Jk^b和抗-Mur。结论患者血清中不规则抗体是导致交叉配血不合的主要原因。     

IgG-Jk~a联合IgG-cE、IgG-Fy~b抗体鉴定及输血策略分析

目的鉴定1例有复杂不规则抗体的标本,并探讨鉴定此类复杂不规则抗体的思路和方法及输血方案。方法对标本鉴定ABO、Rh、Duffy和Kidd等血型,采用盐水方法和间接抗球蛋白方法进行抗体筛选及鉴定抗体类型,采用盐水法、凝聚胺法和间接抗球蛋白方法进行交叉配血。结果患者血型血清学结果为O,CCDee,Jk(a±b+), Fy(a+b-);基因型结果为Jk(a-b+)和Fy(a+b-)。抗体鉴定结合交叉配血试验共检出血清中存在IgG-c、IgG-E、IgG-Fyb、IgG-Jk~a抗体,筛选以上4种抗原均为阴性的供血者与患者进行交叉配血相合后输注。结论对于复杂不规则抗体的鉴定,应当明确鉴定的思路,将多种实验技术和方法进行组合和分析,有效提高实验室解决疑难问题的能力,为临床提供相合血液,保障输血安全。     

抗-JK~a抗体致疑难配血病例分析

河南大学附属郑州颐和医院收治抗-JK~a抗体引起不规则抗体阳性及疑难配血患者1例。常规用微柱凝胶卡式法对患者进行ABO、Rh(D)血型鉴定、直接抗人球蛋白试验、不规则抗体筛查及鉴定、吸收放散试验以及Kidd血型分型,最后用多种方法交叉配血,筛选出相合血液制品进行临床输注。患者血型为O型Rh(D)阳性,直接抗人球蛋白试验结果阴性,不规则抗体鉴定出特异性抗JK~a抗体,Kidd分型结果Jk(a-b+)。鉴定不规则抗体特异性可以为患者筛选合适的供血者,确保临床输血安全。     

1 例骨髓增生异常综合征多次受血患者血清存在抗-M，抗-cE 和抗-Jkb 抗体的鉴定及配血输血策略

目的 通过对临床送检的1 例骨髓增生异常综合征（myelodysplastic syndrome, MDS）多次受血患者的抗筛阳性标本进行鉴定,探讨此类复杂不规则抗体的鉴定思路、方法及输血方案。方法 鉴定标本ABO,Rh,MNS 和Kidd 等血型,应用2 种谱细胞及筛选出的无偿献血者细胞配合吸收放散实验进行抗体筛查、复杂不规则抗体特异性鉴定,采用盐水法、凝聚胺法和间接抗球蛋白方法进行交叉配血。结果 患者血型血清学结果为O,CCDee,M-N+ 和Jk(a+b-),其血清中检出IgM-M,IgG-cE 和IgG-Jkb 抗体,筛选M,c,E 及Jkb 抗原均为阴性的供血者与患者进行交叉配血,配血相合后进行输注。结论 针对多次受血患者产生的复杂不规则抗体,应灵活运用灵敏度高的血清学方法,联合多种谱细胞,准确鉴定出抗体特异性,为临床提供相合的血液,有效保障输血的安全性。     

多次输血产生抗-S联合抗-Wr^a1例

目的分析1例多次输血患者产生抗-S合并抗-Wr^a的血清学及交叉配血特点。方法采用血型血清学方法对标本进行ABO、Rh、MNS、Diego血型鉴定试验,直接抗球蛋白试验,抗体筛查及抗体鉴定试验,抗体效价测定试验。结果经血型血清学试验确认患者血清中存在抗-S合并抗-Wr^a。选择S,Wr^a抗原阴性献血者血样与患者血样进行交叉配血,配血相合,患者输血后无不良输血反应发生。结论抗-S、抗-Wr^a为低频抗体,虽然随机交叉配血相合率高,但仍需做抗体鉴定,提高输血安全性。     

抗球蛋白试验在疑难交叉配血过程中的重要性分析

目的通过对患者输血前血样进行抗球蛋白试验检查,查找导致临床患者配血不合的原因,配合性输注,确保临床输血安全。方法通过不规则抗体筛选试验,检测患者血清中抗体性质。结果 61例交叉配血不合患者抗球蛋白试验结果显示,由温、冷性自身免疫性抗体及冷凝集素影响配血不合30例;ABO血型系统以外不规则抗体同种免疫性抗体31例,由Rh血型系统同种免疫性抗体导致配血不合占大多数,其中与抗-E抗体有关的患者17例,占由同种免疫性抗体引起配血不合的54.84%。结论患者体内产生的ABO血型系统以外不规则同种免疫性抗体或者温、冷性自身免疫性抗体及冷凝集素等几种因素的影响,是造成临床交叉配血不合的主要原因,Rh血型抗原的复杂性和多态性应引起临床的重视,Rh血型同型输注可降低输血不良反应的发生率。     

同种抗体合并模拟同种特异性自身抗体血清学特点分析及配血对策

目的通过对1例同种抗体合并模拟同种特异性自身抗体患者的抗体进行鉴定,了解Rh血型系统中模拟同种自身抗体的血清学特点,以便提供合适的配血策略。方法采用常规血清学方法对患者进行ABO血型、Rh分型鉴定;并完成直接抗人球蛋白试验、抗体鉴定及交叉配血试验;采用患者Rh抗体类似特异性对应抗原阴性的献血者细胞(B型RhccDEE)进行患者血浆吸收放散试验,对吸收后的血浆、红细胞酸放散液进行抗体鉴定。采用患者红细胞放散液稀释技术鉴定抗体特异性。采用盐水法、凝聚胺法、微柱凝胶广谱抗球蛋白卡(LISS/coombs)法、经典抗球蛋白法、酶法、酶-IAT法进行交叉配血试验。结果患者为B型RhccDEE,直抗阳性。血浆抗体鉴定为IgG类的抗-C、e(不排除抗-Ce可能性)、效价为256。对应抗原阴性的献血者细胞吸收血浆后放散液抗体鉴定为IgG类的抗-C、e(不排除抗-Ce可能性)。患者自身红细胞酸放散液8倍稀释后可鉴定为类似抗-C、e特异性(不排除抗-Ce可能性)。对应抗原阴性的献血者红细胞与患者进行主侧配血,经典抗球蛋白法、酶法结果相合;而凝聚胺法、微柱凝胶广谱抗球蛋白卡(LISS/coombs)法、酶-IAT法不合。结论模拟同种自身抗体的正确鉴定对选择适合的交叉配血方法起着至关重要的作用,而筛选出相应抗原阴性的血液输注,可最大程度保证输血的安全、及时、有效。     

红细胞血型系统IgG型不规则抗体固相凝集法检测试剂盒的研发

目的研发基于固相凝集技术的红细胞血型系统IgG型不规则抗体检测试剂盒并评估其性能。方法采用单克隆抗人-RBC抗体包被96孔微板,按100μL/孔加入红细胞悬液形成细胞单层,经过裂解后形成红细胞膜抗原分子单层,加入保护液冷冻干燥后制备成检测用的反应微板;用梯度倍比稀释的多克隆羊抗球蛋白试剂(IgG+C3d/4)与IgG抗-D致敏的O型红细胞反应,筛选构建最佳指示系统;用不同批次的试剂盒检测低效价的IgG抗-D和抗-E,评估试剂盒在保存期内的抗原稳定性;与微柱凝胶抗球蛋白卡、进口同类试剂盒检测不同效价的抗体以测试3者的灵敏度;与进口同类试剂盒对350例血样标本进行检测,以明确2者的检测效能是否一致。结果 20μg/mL的抗人-RBC抗体溶液能够很好的固定红细胞膜抗原分子单层;稀释16倍的多克隆羊抗球蛋白试剂与致敏红细胞构建的指示系统指示效果最佳;试剂盒中冻干的红细胞膜抗原在6个月的保存期内保持稳定;本研究的试剂盒检测灵敏度高于Capture-R Ready Screen试剂盒和抗球蛋白卡;试剂盒与Capture-R Ready Screen对血样标本检测的阳性一致性百分率为98.0%,阴性一致性百分率为99.66%,总一致性百分率为99.43%。结论成功研发了基于固相凝集技术的用于红细胞IgG型同种免疫性不规则抗体检测的试剂盒,该试剂盒具有灵敏度好、保存期长、性能稳定等优势,与进口同类试剂盒等效。     

Capture-R法与微柱凝胶法在不规则抗体筛查中的应用

目的探讨Capture-R法与微柱凝胶法在不规则抗体筛查中的临床应用价值。方法采用微柱凝胶法与Capture-R法对600份临床患者标本做不规则抗体筛查平行试验,对抗体筛查阳性标本进行抗体鉴定。分析比较两种方法在不规则抗体筛查试验中的灵敏度和特异性。结果微柱凝胶法和Capture-R法阳性检出率分别为3.17%（19/600）和2.83%（17/600）,两种方法灵敏度比较差异无统计学意义（P〉0.05）;特异性分别为89.47%（17/19）和100.00%（17/17）,两种方法特异性比较差异无统计学意义（P〉0.05）。结论 Capture-R法在不规则抗体筛查试验中的灵敏度和特异性均达到微柱凝胶法检测水平,可有效检出有临床意义的红细胞抗体,提高临床输血安全。     

血型不规则抗体检测的方法学评价

目的寻找简便、快速、敏感的用于红细胞不规则抗体筛选的试验方法。方法收集6例临床疑难病例样本和1份IgG型试剂,采用盐水法、木瓜酶法、凝聚胺法、抗人球蛋白法、微柱凝胶法平行检测患者血清中不规则抗体的效价,比较5种方法的敏感性、特异性。结果盐水法仅检出IgM类不规则抗体;木瓜酶法、凝聚胺法、抗人球蛋白法、微柱凝胶法在检测IgG类抗体效价时差异不大,木瓜酶法的敏感性略低,微柱凝胶法对弱阳性IgG抗体检测的敏感性最高,但4种方法间差异无统计学意义（P〉0.05）,相关性较好（r〉0.99）。结论盐水法不能检出IgG抗体;木瓜酶法检测IgG抗体具有一定的局限性;凝聚胺法简便、快速,但为手工操作,不易标准化;抗人球蛋白法需要反复洗涤红细胞,费时,不便于开展大规模筛查;微柱凝胶法操作简便迅速、敏感性高、重复性好、易于标准化,适于临床输血前和孕妇产前大批量抗体筛选,是检测IgG抗体最为敏感的方法。     

Activation of human platelets by antibodies to thymocytes and beta 2-microglobulin. III. Effect of cell absorptions on the platelet aggregating and lymphocytotoxic activities of HATG and SA beta 2mG

Horse anti-human thymocyte globulin (HATG) and sheep anti-human beta-2-microglobulin globulin (SA beta 2-mG), both known to have immunosuppressive potency, were quantitatively absorbed by a number of human cells, then, tested for platelet aggregating and lymphocytotoxic reactivities. Peripheral blood lymphocytes, lymphoblastoid T- and B-cell lines, platelets and spermatozoa could remove varying amounts of both reactivities from HATG and SA beta 2mG. Daudi cells were effective in reducing the lymphocyte and some platelet reactivity of HATG but did not affect those of SA beta 2mG. Erythrocytes had no absorption capacity for either antibody. The qualitative and quantitative differences in absorbing with various cells on the platelet aggregating and lymphotoxic activities of HATG and SA beta 2mG indicate the high specificity and direct action of the antibodies on platelets and lymphocytes.     

Pre-transfusion testing problems caused by anti-lymphocyte globulin and their solution

Anti-lymphocyte globulin (ALG) is an antibody to human lymphocytes used to decrease T-cells in renal transplant patients. We recently encountered serologic problems in testing blood from patients treated with ALG. Thirty-nine patients undergoing acute kidney rejection developed positive direct and indirect antiglobulin tests following the administration of equine ALG. Sera from these patients reacted with all red cells (RBCs) tested using both polyspecific and monospecific anti-IgG anti-human sera. Eluates prepared from the patients' RBCs showed similar reactivity. The ALG panagglutinin did not react by manual hexadimethrine bromide (Polybrene) technique. The ALG panagglutinin could be neutralized by anti-human globulin. In our hands, these techniques were useful in distinguishing ALG panagglutinin from co-existing alloantibodies.     

Column agglutination technology: the antiglobulin test

A new system for typing and screening blood, based on the sieving effect of glass bead microparticles, has been developed. The test is performed in a microcolumn in which the red cell agglutinates are trapped in the glass bead matrix during centrifugation, and unagglutinated cells form a pellet at the bottom of the column. Anti- human globulin reagents were incorporated in the diluent and the new test system, column agglutination technology, was compared to conventional tube tests and low-ionic-strength method. Sera and plasmas (228 samples) were screened for red cell antibodies with two anti-human globulin reagents: one containing only anti-IgG and the other containing both anti-IgG and anti-C3b, -C3d. After initial testing, there was 94-percent agreement between column agglutination technology and tube tests, and after repeat testing, there was 97-percent agreement. The column agglutination technology anti-human globulin test eliminates the need to wash red cells, which decreases the overall test time. The test is easy to perform, and the results are more objective than those with tube and microplate methods.     

血型不规则抗体检测的方法学评价

目的寻找简便、快速、敏感的用于红细胞不规则抗体筛选的试验方法。方法收集6例临床疑难病例样本和1份IgG型试剂,采用盐水法、木瓜酶法、凝聚胺法、抗人球蛋白法、微柱凝胶法平行检测患者血清中不规则抗体的效价,比较5种方法的敏感性、特异性。结果盐水法仅检出IgM类不规则抗体;木瓜酶法、凝聚胺法、抗人球蛋白法、微柱凝胶法在检测IgG类抗体效价时差异不大,木瓜酶法的敏感性略低,微柱凝胶法对弱阳性IgG抗体检测的敏感性最高,但4种方法间差异无统计学意义(P>0.05),相关性较好(r>0.99)。结论盐水法不能检出IgG抗体;木瓜酶法检测IgG抗体具有一定的局限性;凝聚胺法简便、快速,但为手工操作,不易标准化;抗人球蛋白法需要反复洗涤红细胞,费时,不便于开展大规模筛查;微柱凝胶法操作简便迅速、敏感性高、重复性好、易于标准化,适于临床输血前和孕妇产前大批量抗体筛选,是检测IgG抗体最为敏感的方法。     

Comparative sensitivity of solid phase versus PEG enhancement assays for detection and identification of RBC antibodies.

Blood banks require a sensitive, specific, and efficient method to detect clinically significant RBC antibodies. Solid phase antibody screening methods are popular due to high sensitivity and automation. However, the high degree of reactivity detects "false positive" antibodies of questionable clinical significance leading to additional testing. We studied positive rates of Capture-R vs. PEG methods and categorized RBC antibodies identified by initial test results of 33,564 consecutive samples by Capture-R method. Capture-R was positive in 1,084/33,564 (3.2%) of samples. Using PEG as our "gold standard", PEG confirmed true positivity (i.e., > or = 1 cell reacting) in 710 Capture-R positive samples (65.5%); 374 Capture-R positive samples (34.5%) did not react in PEG (i.e., false positives). Of the 710 samples with true positivity, only 510 showed clinically significant alloantibodies. Using PEG as our "gold standard", only 2/3 of reactions by Capture-R were considered true positives. Because of ease and automation, Capture-R is popular as a screening test, but a more specific method may be helpful in order to identify truly significant alloantibodies.     

Serologic evidence that factor IX inhibitor in the plasma of hemophilia B patients detects factor IX on normal red cells

BACKGROUND: Patients with hemophilia B lack factor IX (F IX). These patients may become alloimmunized after the transfusion of F IX concentrates and may develop F IX inhibitors, which have been characterized as polyclonal IgG4 alloantibodies. Two cases in which F IX inhibitors caused difficulty in compatibility testing and antibody identification were encountered. It was hypothesized that, because F IX is present in normal plasma, it might be adsorbed by red cells in vivo and then be detected during antibody screening tests with serum containing F IX inhibitors. CASE REPORT: Sera from two African American half-brothers with hemophilia B were incompatible with all common and rare red cell phenotypes tested in the anti-human globulin test, but did not react with each other's red cells. The brothers' red cell antibodies were neutralized with both normal plasma and a commercially available F IX concentrate, which indicated that the red cell incompatibility was most probably caused by their F IX inhibitors. Red cells from an unrelated patient with hemophilia B and a very low titer of F IX inhibitor were tested against the half-brothers' sera and did not react. The compatible red cells from one of the half-brothers and the unrelated patient with hemophilia B adsorbed F IX from normal plasma or F IX concentrate after 37 degrees C incubation; this rendered them incompatible with the plasma containing F IX inhibitor from the other half-brother. CONCLUSION: F IX appears to be present on normal red cells and may be detected during compatibility and antibody identification procedures when serum or plasma containing F IX inhibitors is tested.