Rh阴性孕妇血型D抗原同种免疫反应研究

目的研究Rh阴性孕妇血型D抗原同种免疫反应,为建立有区别的D亚型的输血规则提供参考。方法对RhD阴性孕妇进行基因分型,并检测是否有抗-D;总结分析孕妇妊娠次数以及胎儿红细胞的检出与产生抗-D之间的相关性。结果在122名RhD表型阴性的孕妇中,检出d/d基因型97例,Del型20例,弱D15型1例,RHD-CE(2—9)-D/d融合基因型3例,DⅥⅢ型1例。在97名RhD阴性(d/d)孕妇中,有38例产生抗-D;20名Del血型孕妇均未检出抗-D;3名RhD-CE(2—9)-D/d中有2例产生抗-D;1名弱D15型的孕妇未检出抗-D;1名DⅥⅢ型孕妇抗-D检出阳性。统计分析表明孕妇的妊娠次数以及外周血中检出胎儿红细胞的检出与抗-D的产生有相关性,这可以成为Rh阴性孕妇被D抗原免疫的证据。结论不同D亚型的个体可能会对D抗原有不同的免疫反应,我们认为在中国Del血型的受者可以接收D阳性血液。建立有区别的输血规则可以节约有限的Rh阴性血资源和提高输血安全。     

20例输注D~(el)型红细胞的RhD阴性受者的回顾性分析

目的探讨Del型红细胞是否诱导RhD阴性受者产生同种免疫反应。方法用吸收放散试验鉴定Del型血液,对临床输注Del型红细胞的患者进行追踪,采集输血后1周～1年的患者血液标本,RhD抗原初筛阴性的标本采用间接抗球蛋白试验(IAT)进行阴性确认。对患者血清采用凝聚胺法、间接抗球蛋白试验和微柱凝胶法筛查不规则抗体,然后对筛查阳性标本用谱细胞进行不规则抗体鉴定,并进行效价测定。结果采集到输注Del红细胞的RhD阴性受血者样本20例,男性10例,女性10例,女性均有孕产史。Del型受血者5例,男性2例,女性3例,输注Del红细胞后未发生输血反应,不规则抗体筛查阴性。真实RhD阴性受血者15例,男性8例,女性7例,8例男性和5例女性患者输注Del红细胞后未发生输血反应,不规则抗体筛查阴性;2例妊娠女性抗-D为阳性,未出现输血反应。结论 Del型个体未检出抗-D,产生抗-D的个体未检出Del型。Del红细胞导致RhD阴性受者产生同种免疫反应的概率可能很低。     

RhD阴性患者输注“亚洲型”DEL型红细胞短期内引起高效价抗-D的同种免疫研究

目的 通过对1例RhD阴性患者输注“亚洲型”DEL红细胞后诱发同种抗-D免疫的研究分析,探讨RhD阴性患者输注DEL型策略。方法 采用血清学方法进行ABO及Rh系统血型抗原检测、不规则抗体筛查及鉴定,高分辨熔解曲线和基因测序分析检测DEL型的RHD基因分型。结果 受血者为O型,RhD阴性。供血者为RHD 1227G>A突变的“亚洲型”DEL,R h表型为Ccee。受血者输血后抗体筛查阳性,并鉴定为抗D和抗C,抗D效价512,抗C效价128。结论 RhD阴性患者输注“亚洲型”DEL红细胞会引起抗-D同种免疫。因此对于RhD阴性献血者应加强DEL型的筛查,必要时可以增加RHD基因检测以保障输血安全。     

RhD阴性患者输注RhD阳性同型红细胞血液抗-D检测及其效价测定

目的了解患者Rh D抗原阴性患者在抢救时,输注与患者血型相同的Rh D阳性的血液。患者抗-D产生情况及效价水平。方法 Rh D抗原阴性患者输注Rh D抗原阳性血液后,5、10、20 d连续追踪3次,。使用微柱凝胶抗人球蛋白试剂卡检测患者红细胞不规则抗体及效价[1]。结果 15例Rh D抗原阴性的患者输注同型Rh D抗原阳性红细胞后,共有4例患者血清红细胞不规则抗体筛查阳性[2]。1例女性患者5 d血清红细胞不规则抗体筛查阳性,10 d后又有2例女性患者和1例男性患者血清红细胞不规则抗体筛查阳性。5 d后阳性标本抗体效价为4,20 d后抗体效价结果为16,3例10 d抗体效价为4,20 d后效价为16。结论 Rh D阴性患者输注同型Rh D阳性血液后,机体被所缺失的抗原免疫刺激,产生免疫性抗-D。抗体产生的几率与抗体效价的增高和输注次数及抗原量的多少呈正相关系数上升[3]。     

Rh(D)阴性孕妇血液中胎儿微量Rh(D)红细胞与新生儿溶血病发病率的相关性研究

目的运用流式细胞术产前非侵入性检测Rh(D)血型不合导致新生儿溶血病,探讨该方法在临床的实用性。方法运用已建立的间接标记法(一抗为IgG抗-D,二抗为FITC-抗人IgG(γ),F(ab’)2)染色,采用流式细胞术检测Rh(D)阴性孕妇血液中胎儿微量红细胞。结果流式细胞术与血清学方法检测RhD血型不合导致新生儿溶血病差异不显著。追踪检测Rh(D)阴性孕妇体内胎儿微量Rh(D)红细胞所占比例数据显示,较早进行产前诊断能在一定程度上帮助预防、诊断HDN。结论建立了流式细胞术产前非侵入性检测Rh(D)血型不合导致新生儿溶血病的方法;孕妇在产前常规作流式细胞术检测,为Rh(D)血型不合导致新生儿溶血病提供了一种可靠的诊断依据。     

Rh(D)阴性无偿献血者Rh表型及Del检测结果与分析

目的明确Rh(D)阴性无偿献血者Rh表型及Del型的分布情况,充分掌握Rh(D)阴性献血者资料,为临床需求提供保障。方法采用盐水介质试管凝集法进行Rh(D)初筛,初筛阴性标本再用抗球蛋白法进行Rh(D)阴性确证试验。用生理盐水介质试管凝集法将Rh(D)确证实验阴性无偿献血者标本进行ABO血型和Rh表型检测。采用吸收放散试验对Rh(D)确证阴性无偿献血者标本进行Del型检测。结果筛检并确证1451份Rh(D)阴性无偿献血者,ABO血型分类结果包括A型449份、B型384份、AB型146份、O型472份。性别分类结果包括男性842份、女性609份。Rh(D)检测结果中40份弱D、395份Del阳性、1016份Del阴性。Rh表型中29份CcdEe型、439份Ccdee型、93份CCdee型、778份ccdee型、99份ccdEe型、5份ccdEE型、1份CcdEE型、2份CCdEE型、1份CCd--型、3份--dee型、1份--dEe型。结论本血站具有丰富的Rh(D)阴性无偿献血者,基本满足临床需求;对Rh(D)阴性献血者标本进一步开展Rh表型和Del检测,掌握Rh(D)阴性供血者信息资料,有助于保障Rh(D)阴性受血者的输血安全。     

RHCE(2-9)-D变异产生抗-D引起新生儿溶血病1例

目的 对1例有输血史多次分娩史的RhD阴性孕妇D抗原进行血清学和分子生物学检测,明确RhD表型和基因型。方法 微柱凝胶技术进行母亲和新生儿Rh血型初筛,母亲红细胞经典抗人球蛋白技术进行RhD阴性确认,吸收放散试验确定是否为Del。基因检测确定孕妇RHD基因型。结果 该孕妇结果为Ccdee, RhD确证试验为D阴性,吸收放散试验阴性,基因检测为RHCE(2-9)-D变异,产生抗-D,抗-E。新生儿为CCDee表型,游离抗体检出抗-D和抗-E,放散液检出抗-D。患儿诊断为新生儿溶血病,进行输血、光疗和换血治疗。结论 RHCE(2-9)-D变异型血清学可能表现为D阴性,确证试验阴性,有产生抗-D并引起新生儿溶血病的风险。     

Rh血型系统Del型研究进展

Rh血型系统是人类红细胞血型系统中最为复杂且最具多态性的系统,在临床的重要性仅次于ABO血型系统,而且其分子机理又非常复杂.到目前为止已发现至少49种抗原,其中与临床输血安全密切相关的Rh血型抗原主要有D、C、C、E、e这5种.Rh(D)还存在一些变异体——部分D、弱D和Del等.1984年日本学者Okubo在Rh(D)阴性献血者中首次发现1种非常弱的Rh(D)抗原表达型,命名为Del表型.其非常弱的Rh(D)抗原无法用敏感的间接抗球蛋白试验检出,而只能通过吸收放散试验才可检出.由于人们在Del型是否归于Rh阴性这点上并没有形成共识,故早期Del型献血者血液一直被当作Rh(D)阴性血液而应用于临床输血.     

新疆地区RhD阴性孕妇血清IgG抗-D筛查与效价检测的分析

目的对我院Rh D阴性孕妇IgG抗-D筛查与效价检测的意义进行分析。方法采集1 397例Rh D阴性孕妇的血清,作血型抗体筛查与特异性鉴定,并对抗体阳性患者作效价检测,并追踪抗体阳性患者的新生儿状态。结果在1 397例Rh D阴性孕妇血清中,IgG抗-D阳性146例,阳性率为10.45%(146/1 397),观察146例孕妇IgG抗-D与Rh-HDN发病率的相关性。结论 Rh D阴性孕妇IgG抗-D筛查与效价测定,发现抗体效价16为Rh-HDN发生的临界值,对于Rh血型不合新生儿溶血病的预测、治疗具有重要价值。     

100例RhD初筛阴性孕妇D放散型表型筛查及基因型分析

目的对100例Rh D阴性孕妇进行D放散型(Del)表型筛查及基因型分析,同时进行抗体筛查和鉴定,以指导Del血型孕妇的产前监测。方法 2016年2月—2016年11月期间,收集100例多次妊娠且初筛为Rh D阴性的孕妇外周血标本,采用抗球蛋白凝胶卡法检测Rh D血型,抗球蛋白凝胶卡法进行直接抗球蛋白试验;采用Rh血型分型卡检测Rh CE抗原;抗球蛋白凝胶卡法进行不规则抗体筛查及抗体鉴定。采用吸收放散试验进行Del血型表型鉴定,同时采用序列特异性聚合酶链反应(PCR-SSP)进行RHD*01EL.01(RHD*1227A)等位基因检测,应用限制性片段长度多态性聚合酶链反应(PCR-RFLP)对Del表型标本RHD基因合子型进行进一步分析;对D变异型标本,进行D抗原表位检测(D-Screen)和RHD基因10个外显子的PCR扩增及直接测序分析。结果 100例标本中检出Del表型27例(27%),均没有产生抗-D,其中21例Rh CE抗原分型为Ccee(77.8%),4例为CCee(14.8%),2例为Cc Ee(7.4%);25例RHD基因型为RHD*01EL.01/01N.01(92.6%),2例RHD*01EL.01/RHD*01EL.01(7.4%)。此外还检出D变异型4例(4%),其中包括2例部分DⅥ3,1例弱D25和1例弱D15,均未产生抗-D。检出Rh D真阴性血型69例,其中产生抗-D 9例(12.7%),且有2例合并抗-C;产生抗-c E合并抗-Jkb1例。结论携带有RHD*01EL.01等位基因的"亚洲型"Del血型孕妇在Rh D阳性胎儿的免疫刺激下,很可能不会产生抗-D,属于完全性Del(complete Del)血型。     

Rhesus alloimmunization following intensive plasma exchange

A 17-year-old woman was admitted to the hospital for the treatment of rapidly progressive systemic lupus erythematosus. She failed to improve when treatment with cyclophosphamide and prednisone and, therefore, was treated with intensive plasma exchange. A total of 24 liters of plasma was exchanged during six separate procedures over an 8-day period. The patient, who was blood group B Rh negative (Cde/cde), was found to have an IgG anti-D antibody reacting at a titer of 16 by the indirect antiglobulin technique 6 weeks after the first plasma exchange procedure. The titer of this antibody subsequently rose to 512. This patient, who had neither been pregnant nor received any blood products other than the plasma used during the plasma exchange, was presumably immunized by Rh positive red cells or stroma present in the transfused plasma. It is estimated that the patient received approximately 10(10) Rh positive cells, or approximately one ml of packed red cells–a quantity sufficient to cause Rhesus alloimmunization.     

酸放散方法对弱D抗原的筛选研究

目的 目的确定用盐水抗D检测为D阴性的红细胞标本中是否存在Del型红细胞标本。方法 将待检红细胞样本与人血清抗D抗体混合吸收。采用酸放散方法将致敏在红细胞上的抗D抗体放散下来，用微柱凝胶抗人球蛋白试剂卡及试管法检测放散液中是否存在抗体。结果 51份由盐水抗D确定为D阴性的红细胞样本中有3人用传统试管法抗人球蛋白实验检测为弱凝集；经酸放散实验后，检测放散液，此3人放散液中均存在较强的抗D抗体，以此确定此3人红细胞抗原为Del型。结论 酸放散实验在临床输血中可做为确定红细胞上是否存在弱D抗原的敏感方法。     

Complement fixation by Rh blood group antibodies

Rh blood group antibodies normally do not fix complement. Rh positive intact red blood cells treated with papain do not lyse when incubated with corresponding antibody and complement. This study was done to determine if complement fixation occurs when antibodies were combined with Rh positive red blood cell ghosts untreated or treated with papain. Complement fixation was observed with IgG anti-D, anti-DC and anti-c when papain treated ghosts were used. No complement fixation or a smaller degree of it was observed in the case of untreated Rh positive red blood cell ghosts when incubated with similar anti-Rh antibodies. It is concluded that the papain treatment of Rh positive red blood cell ghosts, possibly by inducing aggregation of antigen sites, allowed complement binding by Rh antibodies.     

Hemotherapy in patients undergoing blood group incompatible bone marrow transplantation

The management of hemotherapy in 31 cases of ABO- or Rh-incompatible bone marrow transplantation is described. Our experience confirms that ABO or Rh incompatibility does not adversely affect engraftment, patient survival, or incidence of graft-versus-host disease. Eighteen recipients with ABO antibodies against the donors' red cells (major incompatibility) were managed by different combinations of plasma exchange, transfusion of incompatible donor type red cells, and removal of donor-type red cells from the bone marrow before transplant. The only serious complication was delayed hemolysis in seven of nine patients who received incompatible red cell transfusions before transplant. Thirteen patients received bone marrow containing ABO antibodies against their red cells (minor incompatibility). Five were managed by centrifuging the bone marrow to remove plasma and reduce the amount of antibody. This did not cause substantial loss of stem cell activity (60-100% of original marrow), and no patients had complications related to the marrow transfusion. In contrast, two of seven patients who received uncentrifuged bone experienced hemolysis. Two of four Rh positive recipients who received marrow from an Rh negative donor developed anti-D, possibly due to Rh positive blood components transfused after transplantation. None of eight Rh negative patients who received an Rh positive transplant has developed anti-D. Blood components should be selected to avoid transfusion of incompatible red cells and to avoid transfusion of a large amount of incompatible plasma. This may necessitate use of plasma components of a different ABO type than the red cell components.     

Differentiation of anti-D, -C, and -G: clinical relevance in alloimmunized pregnancies

BACKGROUND: The differentiation of anti-D, -C, and -G specificities is seldom considered clinically important in pretransfusion testing. However, distinguishing these antibody specificities in alloimmunized pregnancies may be essential. The clinical prognosis as well as Rh immune globulin prophylaxis depends on the accurate identification of these antibodies. CASE REPORT: A pregnant woman, para 1 gravida 4, who had received Rh immune globulin at appropriate intervals during her previous pregnancies was reported to have anti-D (titer = 4) and anti-C (titer = 32). Differential adsorption and elution studies showed that the patient had anti-C and anti-G, but not anti-D. This case prompted retrospective examination of the sera from six other women with anti-D and anti-C who were referred to a high-risk pregnancy clinic. Of six pregnant women reported to have anti-D and anti-C; two had anti-D, -C, and -G; three had anti-D and -G, but not anti-C; and one had anti-C and -G, but not anti-D. This last is similar to the index case. CONCLUSION: Cases of pregnant women with anti-C and -G, but not anti-D, are not infrequent. Studies to differentiate anti-D, -C, and -G should be performed on alloimmunized pregnant women presumptively identified as having anti-D and anti-C when the medical history (Rh immune globulin prophylactic therapy) and/or titer values (e.g., anti-C titer higher than anti-D titer) suggest that anti-D may not actually be present. Rh immune globulin has not failed in these patients, and they should receive this therapy during pregnancy to prevent immunization to D.     

The routine use of Rh-negative reagent red cells for the identification of anti-D and the detection of non-D red cell antibodies

BACKGROUND: Before 1987, fewer than 50 patients per year at the authors' laboratory had a positive antibody detection test due to antepartum Rhesus immunoprophylaxis. However, after 1987, a marked increase was observed in the number of patients who had received Rh immune globulin (RhIG) during pregnancy as part of routine antepartum Rh immunoprophylaxis. In anticipation that an increased use of RhIG during pregnancy would increase the number of patients in whom anti-D was detected by this laboratory, a protocol was developed to abbreviate the process required to identify anti-D. Although this protocol was adopted primarily to address an anticipated increase in antenatal RhIG usage in women, it was also applied to alloimmunized Rh-negative males. STUDY DESIGN AND METHODS: When an Rh-negative patient (male or female) had a reactive screening test for unexpected antibodies and met certain other criteria, the patient's serum was tested with a three-vial set of Rh-negative reagent red cells (Rh-negative screening RBCs), instead of with panels of typed RBCs (panel RBCs), for the identification of anti- D or the detection of non-D antibodies. If the serum under test did not agglutinate or hemolyze Rh-negative screening RBCs, anti-D was identified and no further testing was performed. If the serum agglutinated or hemolyzed Rh-negative screening RBCs, conventional testing with panel RBCs was done to determine the antibody specificity. RESULTS: Rh-negative patients (n = 1174) who had reactive screening tests for unexpected antibodies were tested with Rh-negative screening RBCs; 1079 were found to have anti-D as a single antibody. Seven of these patients subsequently developed a non-D alloantibody, after transfusion or pregnancy, and one patient had anti-C that escaped detection at the time of initial testing with Rh-negative RBCs (a false- negative result). Ninety-two patients had anti-D in combination with a non-D antibody, and three patients had a non-D antibody but not anti-D. Use of the anti-D identification protocol actually reduced the laboratory workload by 176 College of American Pathologists workload units per month, in spite of a marked increase in the number of patients in whom anti-D was detected. No hemolytic transfusion reaction was attributed to the abbreviation of anti-D identification. CONCLUSION: The identification of anti-D may be abbreviated without jeopardizing patient safety. Such a protocol can reduce laboratory workload and might be particularly appealing to health care facilities that perform antibody detection testing on large numbers of Rh-negative pregnant women, especially if antepartum RhIG is administered routinely.     

D~(el)红细胞膜表面D抗原表位及抗原强度分析

目的分析Del红细胞膜表面D抗原表位并测定其抗原强度。方法采用16种针对D抗原不同表位的单克隆抗体,通过吸收放散方法检测74例Del个体红细胞膜表面D抗原表位,应用流式细胞术检测其相对抗原强度。结果 74例Del个体D抗原1~9表位检测均为阳性,D阴性、Del及D阳性红细胞相对D抗原阳性率分别为(0.97±0.18)%、(3.06±0.31)%和(99.65±0.71)%,3种表型红细胞D抗原平均荧光强度分别为13.89±2.45,24.18±3.38,223.32±13.05。结论 Del红细胞膜表面D抗原表位表达基本完整,其D抗原强度极低,仅含有微量的抗原分子数目。     

抗-D抗体的定量测定与效价测定的比较性研究

目的Rh系统血型同种抗体是造成严重新生儿溶血疾病的最主要因素。在发达国家和地区,RhD同种抗体监测通常使用抗体定量方法,而在我国通常采用效价测定方法。本研究针对效价测定法与抗体定量法的相关性进行比较,研究效价测定法是否可以真实反映抗体效力。方法本研究中,对17份效价大于64的抗-D血浆样本的效价与抗体定量法进行比较研究,效价法采用ORTHOVISION自动血型检测仪进行自动凝胶柱法检测,抗体定量采用自动连续流抗体定量仪(WHS-D0120)进行检测。结果通过研究,本研究团队发现,抗体效价法与抗体定量法并不是总一致,其相关性为r=0.92(P<0.001),通过对n≥3的效价组(256组、512组、2048组)的比较研究发现,256组与512组间差异有统计学意义(P<0.001),256组与2048组间及512组与2048组间差异无统计学意义(P>0.05),通过单核细胞单层实验,发现效价为256的四份抗体血浆吞噬率组存在较大差异。结论对RhD阴性的患者及孕妇血浆的抗-D评估及监测,抗体定量法较效价法更为精确。     

Immunospecific red cell binding of iodine 125-labeled immunoglobulin G erythrocyte autoantibodies

The primary interaction of autoantibodies with red cells has been studied by using labeled autoantibodies. Immunoglobulin G red cell autoantibodies obtained from IgG antiglobulin-positive normal blood donors were labeled with radioactive iodine and compared with alloanti-D with respect to their properties and binding behavior. Iodine 125-labeled IgG autoantibody migrated as a single homogeneous peak with the same relative mobility as human IgG on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The isoelectric focusing pattern of labeled autoantibodies varied from donor to donor but was similar to that of alloanti-D, consisting of multiple IgG populations with isoelectric points in the neutral to alkaline range. 125I-autoantibody bound to all human red cells of common Rh phenotypes. Evidence for immunospecific antibody binding of the labeled autoantibody was based on variation in equilibrium binding to nonhuman and human red cells of common and rare phenotypes, enhanced binding after red cell protease modification, antiglobulin reactivity of cell-bound IgG comparable to that of cell-bound anti-D, and saturation binding in autoantibody excess. Scatchard analysis of two 125I-autoantibody preparations yielded site numbers of 41,500 and 53,300 with equilibrium constants of 3.7 and 2.1 X 10(8) L X mol-1. Dog, rabbit, rhesus monkey, and baboon red cells were antigen(s) negative by quantitative adsorption studies adsorbing less than 3% of the labeled autoantibody. Reduced ability of rare human D--red blood cells to adsorb the autoantibody and identification of donor autoantibodies that bind to Rh null red blood cells indicated that eluates contained multiple antibody populations of complex specificities in contrast to anti-D, which consists of a monospecific antibody population. Another difference is that less than 70% of the autoantibody IgG was adsorbed by maximum binding red blood cells as compared with greater than 85% for alloanti-D. Evidence obtained indicates that the red blood cell nonadsorbable IgG, some 30%, consists of other IgG populations, one of which represents IgG autoanti-idiotypes to the autoantibody.     

Rh 新生儿溶血病的血清抗体分析

目的对20例Rh新生儿溶血病血清抗体回顾性分析。方法采用直接抗人球蛋白试验、游离抗体试验、放散试验检测ABO以外的抗体,采用微柱凝胶间接抗人球蛋白试验进行抗体鉴定。结果20例Rh溶血病患儿中由抗D引起的溶血病有11例,由抗-D和Rh其他系统抗体联合引起的有4例,共15例,占75%;由抗-E引起有3例,由抗-E和抗-c联合引起1例,占20%;由抗-C引起1例,占5%。20例患儿母亲都曾有生产或流产或输血史。结论为预防新生儿Rh溶血病的发生,尤其是对曾有过生产史、流产史或输血史的孕妇作产前夫妇Rh血型和孕妇Rh免疫性血清抗体筛查极有必要。