An acute hemolytic transfusion reaction caused by an anti-P1 that reacted at 37 degrees C

BACKGROUND: Hemolytic transfusion reactions (HTRs) due to anti-P1 have rarely been reported. There is only one report (from 1945) of an acute HTR due to anti-P1. CASE REPORT: A 74-year-old woman with anti-P1 was given blood that had been found to be compatible by the use of prewarmed serum and saline-suspended red cells (RBCs) and of an antiglobulin test with anti-IgG. The test mixtures were not centrifuged or inspected for agglutination after the 37 degrees C incubation phase. After transfusion of 50 mL of P1 + blood, the patient had an acute HTR (hemoglobinemia, hemoglobinuria, and increased blood pressure, temperature, and respiration). RESULTS: When studied by a reference laboratory, the anti-P1 was shown to be easily detectable (3+ agglutination) by a prewarming technique (saline or low-ionic-strength saline [LISS]), which included centrifugation at 37 degrees C, but only weak reactions were observed when centrifugation after 37 degrees C incubation was omitted. The indirect antiglobulin test was weakly positive (1+) with anti-IgG, but polyspecific anti-human globulin reacted 2+. The anti-P1 agglutinin was IgM, and its titer was 16 at 37 degrees C (prewarmed) and 256 at 23 degrees C; it caused hemolysis of RBCs at 37 degrees C under conditions known to enhance hemolysis. An indirect monocyte monolayer assay gave results of 11.2 and 22 percent in testing of P1 + RBCs incubated with the patient's serum alone and with patient's serum plus fresh normal serum (as a source of complement), respectively (normal < or = 3%). CONCLUSION: An acute HTR was caused by a hemolytic anti-P1 that reacted at 37 degrees C. This antibody was not detected by the hospital in a prewarmed crossmatch that omitted 1) the addition of LISS, 2) the reading for agglutination after the 37 degrees C incubation, and 3) the use of antiglobulin sera containing anti-complement.     

False-positive reactions of some monoclonal anti-c reagents

Cross-reactivities of monoclonal reagents for red blood cell (RBC) typing are seen very rarely, e.g. in some reagents for testing of ABO or Lewis antigens. We report on two patients in whom we observed weak reactions using a monoclonal anti-c reagent containing clone MS35, but no reactions with anti-c reagents containing other monoclonal or polyclonal antibodies. To resolve this discrepancy, we studied RHCE exon 1 (Ex1) and 2 from genomic DNA of one patient and performed quality controls of the false-positive reacting anti-c reagent. RHCE Ex1 showed no abnormalities as well as RH Ex2 examined with primers specific for Ex2 D/C. No amplification product of RH Ex2 was received with primers specific for c. A titration study with untreated, papain-treated and sialidase-treated adult and newborn RBCs showed anti-i reactivity of the false-positive reacting reagent. This reactivity could not be removed by absorption with c-negative newborn RBCs without reducing the anti-c titre in the same way, indicating a cross-reactivity. Some manufacturers give a remark on a possible cold agglutinin activity of their monoclonal anti-c reagents in the instruction leaflet, but in order to avoid such irritation, we recommend to remove this cross-reactivity by dilution during the manufacturing process.     

IMMUNOHEMATOLOGY: Novel antibody screening cells,MUT+Mur kodecytes,created by attaching peptides onto red blood cells

BACKGROUND: Antibody screening and identification panels are generally limited by the natural antigenic phenotypes present in their source donor population. However, the recent ability to attach peptides to the surface of cells has opened up the opportunity to create red blood cells (RBCs) with antigen profiles specifically designed for antibody screening and identification in a target population. STUDY DESIGN AND METHODS: Clinically significant antibodies to variant glycophorins (GPs) such as GP.Mur are more commonly seen in certain Asian populations. Using peptides representative of the MNS antigens MUT and Mur, RBC antibody screening cells were created using KODE cell surface engineering constructs. MUT‐, Mur‐, and MUT+Mur‐modified RBCs, known as kodecytes, were tested against monoclonal reagents and polyclonal sera with specificity for epitopes on GP.Mur‐positive RBCs. RESULTS: Kodecytes retained their normal expression of intrinsic blood group antigens while expressing the new epitopes attached by KODE technology. The MUT, Mur, and MUT+Mur kodecytes, although unreactive with the various monoclonal reagents, were appropriately reactive with polyclonal sera containing antibodies reactive with GP.Mur‐positive RBCs. CONCLUSIONS: This study used selected MUT and Mur peptides and KODE cell surface engineering technology to create MUT+Mur kodecytes suitable for the detection and identification of RBC antibodies in human serum or plasma. This technology has the potential to create a large range of specialized RBCs for antibody screening and identification.     

A newly developed gel centrifugation test for quantification of RBC-bound IgG antibodies and their subclasses IgG1 and IgG3: comparison with flow cytometry

BACKGROUND: Novel gel centrifugation test (GCT) cards were evaluated with respect to their ability to estimate the quantity of IgG on RBCs and the determination of the IgG subclasses IgG1 and IgG3. STUDY DESIGN AND METHODS: In 65 patients with a positive DAT, the amount of IgG-gamma-, IgG1, and IgG3 on RBCs was examined by use of GCT cards and flow cytometry (FC) in parallel. The results were correlated with the presence or absence of hemolysis. In addition, D+ RBCs were studied after sensitization with anti-D sera from 22 alloimmunized pregnant women. RESULTS: The amount of IgG on the RBCs as determined by GCT dilution cards correlated with FC (r=0.70, p < 0.0001). IgG subclass results as determined by GCT IgG subclass cards were confirmed by FC in 14 cases with an anti-IgG-gamma-chain titer > or =300, whereas IgG subclass cards were not suitable in cases with anti-IgG-gamma-chain titers less than 300. In 44 patients with 2+ or 3+ DAT in the GCT and anti-IgG-gamma-chain titer < or =30, no hemolysis was observed, whereas hemolysis occurred in 13 of 14 patients with an anti-IgG-gamma-chain titer > or =300. GCT data obtained by IATs with anti-D sera were concordant with FC results. CONCLUSION: There is a correlation between the amount of RBC-bound IgG and immune hemolysis. The GCT cards that detect the anti-IgG-gamma-chain may be useful to predict hemolysis in patients with a 2+ or 3+ DAT in the GCT. The diagnostic value of GCT cards for IgG subclass testing should be investigated further.     

Tazobactam-induced haemolytic anaemia, possibly caused by non-immunological adsorption of IgG onto patient's red cells

A patient with pneumonia was treated with Tazocin (piperacillin/tazobactam). However, the expected haemoglobin (Hb) increment after transfusion was not achieved. Plasma bilirubin and lactate dehydrogenase were raised. The direct antiglobulin test (DAT) was positive (4+) for immunoglobulin G (IgG) only, but no RBC antibodies were demonstrable in the plasma or an eluate from the patient's RBCs. Drug-induced haemolysis was suspected. After discontinuing Tazocin administration, Hb and bilirubin levels returned to expected values. The patient's plasma gave a positive (3+) indirect antiglobulin reaction only with RBCs pretreated with tazobactam. However, random patient plasmas also gave weak (+/- to 1+) reactions, indicating non-immunological adsorption of IgG onto RBCs rather than specific anti-tazobactam antibodies. Subsequently, plasma samples with varying IgG levels (0.8-89.7 g L(-1)) were tested against RBCs pretreated with tazobactam. The amount of plasma IgG non-immunologically adsorbed onto the drug-coated RBCs was found to correlate directly with the plasma IgG level. The patient had a high plasma IgG level (41.6 g L(-1)) which explains why the antiglobulin test was stronger with the patient's plasma than with random plasma samples. Previous reports (Garratty & Arndt, (1998) British Journal of Haematology, 100, 777-783; Arndt & Garratty (2000) Transfusion, 40, 29S) suggested that non-immunological coating of RBCs with IgG may affect RBC survival; our results would support that suggestion. This is the first reported case of haemolytic anaemia associated with tazobactam.     

False-positive eluate reactivity due to the low-ionic wash solution used with commercial acid-elution kits

BACKGROUND: During the use of commercial red cell (RBC) acid-elution kits for adsorption and elution (adsorption/elution) studies with anti- D, unexpected reactive eluates (anti-D) were obtained from D- RBCs. Such results were not obtained with a parallel xylene method or, historically, with heat and ether methods. STUDY DESIGN AND METHODS: Single-donor and commercial polyclonal anti-D samples were incubated with D+ and D- RBCs. Acid eluates were prepared by the manufacturers' directions. Variations in the wash step of the eluate preparation included the use of commercial kit wash solution versus phosphate- buffered saline versus solutions of various ionic strengths. RESULTS: Anti-D was eluted from 20 of 22 samples of D- RBCs after incubation with commercial polyclonal anti-D (titer 512) and from 2 of 3 samples of D- RBCs incubated with single-donor anti-D (titer 256). With a low- titer (16) single-donor anti-D, 0 of 4 eluates from D- RBCs reacted. When phosphate-buffered saline was substituted for the commercial wash solution, 0 of 11 D- RBC eluates reacted, as compared with 9 of 11 D- RBCs that yielded positive 1+(-)2+ eluates with the commercial wash solution. If the recommended initial phosphate-buffered saline wash was omitted before the use of the commercial wash solution, the eluate reactivity was stronger (2+(-)3+). When low-ionic-strength (< 0.03 M) saline was substituted, anti-D was eluted from D- RBCs. All last washes were nonreactive. Antiglobulin tests on all adsorbing D- were negative. CONCLUSION: Commercial wash solutions used for acid elution are at low ionic strength and commonly yield superior eluates, but in the presence of high-titer antibodies, false-positive eluates can result. It is our belief that the low-ionic-strength wash solution caused aggregation of IgG and nonspecific attachment of IgG on RBCs. Aggregates will contain IgG serum antibodies in proportion to the titer of the antibody. It is this nonspecifically bound antibody that is eluted from antigen- negative RBCs.     

Serology of antibodies to second- and third-generation cephalosporins associated with immune hemolytic anemia and/or positive direct antiglobulin tests

BACKGROUND: First-generation cephalosporins rarely caused immune hemolytic anemia (IHA). Second- and third-generation cephalosporins, especially cefotetan and ceftriaxone, are increasingly associated with severe, sometimes fatal IHA. STUDY DESIGN AND METHODS: Samples from 53 patients with drug-induced IHA and/or positive direct antiglobulin test (DAT) were tested. Patients' sera were tested against drug-treated red cells (RBCs) and untreated or enzyme-treated RBCs, with and without the addition of drug solution. Eluates from patients' RBCs were tested against drug-treated and untreated RBCs. RESULTS: Forty-three patients had antibodies to cefotetan, 8 to ceftriaxone, 1 to cefoxitin, and 1 to cefotaxime. All patients had a positive DAT; only anticefoxitin and anti-cefotetan were demonstrable in RBC eluates. Sera containing anti-cefoxitin, anti-cefotaxime, and anti-cefotetan reacted with drug-treated RBCs (100%) and untreated or enzyme-treated RBCs in the presence of drug (98% or 100%, respectively). All of the ceftriaxone antibodies reacted with untreated or enzyme-treated RBCs in the presence of drug, but those tested did not react with ceftriaxone-treated RBCs. In addition to cefotetan-dependent antibodies, 19 (44%) and 14 (33%) of 43 sera contained drug-independent antibodies when tested with and without the presence of a polyethylene glycol potentiator, respectively. CONCLUSION: Cefotetan is by far the most common cause of drug-induced IHA. All cefotetan antibodies and the single examples of cefoxitin and cefotaxime antibodies reacted with drug-coated RBCs, and most, in contrast to the reactions of antibodies to first-generation cephalosporins (e.g., cephalothin), also reacted with RBCs (not treated with drug) in the presence of the drug. Ceftriaxone antibodies reacted only by the latter mechanism. Drug-independent antibodies (i.e., those reacting without any drug being present) were detected in 33 to 44 percent of patients' sera containing cefotetan antibodies, depending on the sensitivity of the method used.     

A new test useful in identifying red cells with a (delta-alpha) hybrid sialoglycoprotein

St(a+) and Dantu+ red cells (RBCs), treated with 0.1 percent ficin solutions, reacted strongly with Vicia graminea lectin (anti-NVg). No other RBC sample tested gave these results. This test (ficin-pretreated RBCs tested with anti-NVg [FT-NVg]) was used to screen the RBCs of 300 Oriental and 100 black donors. Three percent of the Oriental donor samples tested were FT-NVg+, whereas no FT-NVg + RBCs were found in the black donor samples tested. The FT-NVg test is easy to perform and gives selective reactions with antigens associated with (delta-alpha) hybrid sialoglycoproteins, such as Sta and Dantu. This test can be used for mass screening of RBCs in a search for St(a+) or Dantu+ RBCs. Another application of the FT-NVg test would be to screen RBCs suspected of having a low-incidence antigen before using scarce anti-Sta and -Dantu reagents.     

Transfusion support with RBCs from an Mk homozygote in a case of autoimmune hemolytic anemia following diphtheria-pertussis-tetanus vaccination

BACKGROUND: Autoimmune hemolytic anemia (AIHA) in children, although unusual, is often associated with recent infection. Several reports have identified the diphtheria-pertussis-tetanus (DPT) vaccination as a possible trigger for AIHA. STUDY DESIGN AND METHODS: Life-threatening AIHA was diagnosed in a 6-week-old infant 5 days after receiving a DPT vaccination. The patient required daily transfusion and/or exchange transfusion for 3 weeks. RBCs from an Mk homozygote were found compatible with the patient's autoantibody. Transfusion of RBCs from an Mk homozygote and later RBCs from an individual (K.T.) with a variant glycophorin, Mi.VII, were required to sustain the patient's Hb level until autoantibody production ceased, as evidenced by a fall in antibody titer and the patient's Hct returning to normal. RESULTS: The DAT was positive (3+) with only anti-C3 on presentation. An IgM cold reactive autoantibody with probable anti-Pr specificity and high thermal amplitude (37 degrees C) was identified in the serum. The DAT was no longer positive after transfusion with compatible blood. CONCLUSION: This case represents life-threatening AIHA in an infant, temporally related to a DPT injection and responsive to a combination of immunosuppression and transfusion of rare compatible blood.     

Hemolytic anemia associated with injection of fluorescein

A 49-year-old woman presented with a hemoglobin level of 9.5 g per dL (95 g/L), reticulocyte count of 6.7 percent (0.067), and hemoglobinuria. The next day, the hemoglobin had dropped to 5.8 g per dL (58 g/L), and total bilirubin was 8.8 mg per dL (150 mumol/L). The serum reacted 2+ with all red cells (RBCs). The direct antiglobulin test (DAT) was 3+ with anti-IgG and 1+ with anti-C3, but eluates prepared by two different methods did not react with untreated RBCs. The eluate reacted 2+ with amoxicillin-coated RBCs; amoxicillin had been listed in the patient's record as a previous medication. The patient denied recent ingestion of amoxicillin. Further investigation documented the injection of a dye, fluorescein sodium (AK-FLUOR-25%), for a ophthalmologic fluorescein angiographic study 2 days before admission. RBCs coated with AK-FLUOR reacted with the eluate. Controls consisting of normal serum, an eluate prepared from DAT-negative RBCs, and a serum known to contain anti-penicillin did not react with AK- FLUOR-coated RBCs. Nine days later, the DAT was negative and the serum did not react with untreated RBCs. In the presence of AK-FLUOR (1-in- 125) or amoxicillin (1 mg/mL), the serum reacted 2+ in the antiglobulin test. Antibodies to AK-FLUOR and amoxicillin appeared to react by two mechanisms, which is similar to results in recent reports of other drugs associated with hemolytic anemia. AK-FLUOR has not previously been reported to be associated with hemolytic anemia.     

Acute intravascular hemolysis secondaryto out-of-group platelet transfusion

BACKGROUND: Acute intravascular hemolysis is rarely associated with platelet transfusion. Out-of-group single-donor platelets may cause hemolysis if the donor has high-titer ABO hemagglutinins. CASE REPORT: A 44-year-old woman, blood group A, was recently diagnosed with acute myeloid leukemia and was receiving chemotherapy. After the transfusion of apheresis platelets from a group O donor, back pain, hemoglobinuria, and hemoglobinemia developed, and her Hb dropped by 2.3 g per dL, despite the transfusion of 2 units of RBCs. RESULTS: Investigation revealed acute intravascular hemolysis with a positive DAT due to anti-A(1) on her RBCs. The donor's titer of anti-A(1) was greater than 16,000. CONCLUSION: Review of published cases raises the possibility that hemolytic reactions to out-of-group platelets may be more frequent since the use of apheresis platelets has increased.     

Monoclonal antibodies in blood group serology

The state of the art of monoclonal antibody specifications and blending characteristics needed to produce high quality reagents are described for ABO, RhD and polyspecific anti-human globulin (AHG) reagents.Some interesting advances have been made. Thus some murine monoclonal anti-As (e.g. MH04) see Ax. Also, they may show feeble reactions with the traces of A on some group B cells (the B(A) phenomenon) and the quality control of these reagents is discussed. The use of anti-A,B reagents is now debatable as monoclonal anti-A/anti-B reagents are now more reliable than conventional reagents for detecting weak sub-groups of A and B.RhD typing by saline routine and rapid tests can now be performed with IgM monoclonal anti-D reagents (e.g. MAD-2), but the old problem of Du and D variants is discussed as IgM anti-D is not reliable for their detection.Monoclonal anti-C3c/C3d or just selected anti-C3d (BRIC-8) blended with rabbit anti-IgG make excellent potent clean AHG reagents. The essential false positive test of fresh serum incubated with CPD-A1 red cells from donor pack segment lines is essential for the adjustment and evaluation of AHG reagents.Selected monoclonal anti-M, anti-N, anti-Le^a and Le^b are established as excellent typing reagents and monoclonal antibodies have now been developed to e, E, K, k and other red cell antigens.     

Murine monoclonal anti-s and other anti-glycophorin B antibodies resulting from immunizations with a GPB.s peptide

BACKGROUND: The blood group antigens S and s are defined by amino acids Met or Thr at position 29, respectively, on glycophorin B (GPB). Commercial anti-s reagents are expensive to produce because of the scarcity of human anti-s serum. Our aim was to develop hybridoma cell lines that secrete reagent-grade anti-s monoclonal antibodies (MoAbs) to supplement the supply of human anti-s reagents.
STUDY DESIGN AND METHODS: Mice were immunized with the GPB^s peptide sequence TKSTISSQTNGE T GQLVHRF. Hybridomas were produced by fusing mouse splenocytes with mouse myeloma cells (X63.Ag8.653). Screening for antibody production was done on microtiter plates by hemagglutination. Characterization of the MoAbs was done by hemagglutination, immunoblotting, and epitope mapping.
RESULTS: Eight immunoglobulin G MoAbs were identified. Five antibodies are specific by hemagglutination for s and two MoAbs, when diluted, are anti-S–like, but additional analyses shows a broad range of reactivity for GPB. Typing red blood cells (RBCs) for s from 35 donors was concordant with molecular analyses as were tests on RBCs with a positive direct antiglobulin test (DAT) from 15 patients. The anti-s MoAbs are most reactive with peptides containing the ³¹QLVHRF³⁶ motif, with ²⁹Thr. By Pepscan analyses, the anti-S–like MoAbs reacted within the same regions as did anti-s, but independently of ²⁹Met. One antibody was defined serologically as anti-U; however, its epitope was identified as ²¹ISSQT²⁵, a sequence common for both GPA and GPB.
CONCLUSION: In addition to their value as typing reagents, these MoAbs can be used to phenotype RBCs with a positive DAT without pre-test chemical modification.     

Invalidation of antiglobulin tests by a high thermal amplitude cryoglobulin

A multiply transfused patient was referred for evaluation of a transfusion reaction. The direct and indirect antiglobulin tests (DAT, IAT) for alloantibody were negative. However, IgG-coated control cells failed to agglutinate in the negative reactions, casting doubt on their validity. At 4 degrees C, the patient's serum exhibited a large cryoprecipitate (2.9 mg/mL), made up predominantly of an IgG kappa paraprotein and having trace amounts of IgM and C3. Clear serum separated at 37 degrees C became cloudy within 10 minutes at room temperature (RT); within 4 hours, approximately 60 percent of the total precipitable cryoprotein had precipitated. Red cells (RBCs) incubated in fresh serum that had cooled to RT or RBCs obtained from RT or refrigerated samples contained cryoprecipitate that sedimented with the RBCs during washing with RT saline. On resuspension, enough IgG cryoglobulin redissolved to neutralize completely the commercial anti-IgG reagents. If the patient's samples were maintained at 37 degrees C, cryoprecipitate did not form, and RBCs washed four times at 37 degrees C gave valid DAT and IAT reactions. The removal of all cryoprecipitate from the patient's serum by centrifugation after overnight incubation at 4 degrees C also made possible valid antibody screening and compatibility tests.     

Two cases of "hrB-like" autoantibodies appearing as alloantibodies

Two cases are described in which autoantibodies mimicked alloantibodies. The direct antiglobulin test (DAT) on the red cells (RBCs) from both patients was negative when routine manual hexadimethrine bromide (Polybrene) and enzyme-linked antiglobulin techniques were used. The RBCs also did not react on direct bromelin and direct Polybrene tests. However, an "hrB-like" antibody was eluted from the RBCs of both patients. The sera from these patients reacted with all e+ hrB+ RBCs but not with e+ hrB-, e-, or their own RBCs. The antibody in the serum of one patient was not adsorbed by R2R2 RBCs. Serologic tests initially suggested (by direct testing and adsorption studies) that the serum antibodies were alloantibodies rather than autoantibodies. RBCs taken from one patient, 8 months after her sample was first referred to our laboratory, reacted with a serum sample from her first admission. An RBC sample taken from the other patient, initially typed e+ and hrB- but 1 month later typed e+ and hrB+ by using the same anti-hrB sera, was used to test the earlier samples.     

单克隆试剂导致ABO定型异常结果的研究

目的 探讨单克隆试剂导致ABO血型鉴定的异常结果。方法 采用单克隆抗-A(B)试剂和人源抗-A(B)血清平行检测受检者红细胞，同时用反定型试验验证。对出现ABO血型异常结果者，再用吸收放散等试验确证。结果 在常规工作中，单克隆试剂导致ABO血型鉴定结果异常者5例。最后被确定为假阴性3例，假阳性2例。结论 对病人的输血前检查，必须增加ABO反定型试验。     

Detection of multiple passively acquired alloantibodies following infusions of IV Rh immune globulin

BACKGROUND: Passively acquired blood group alloantibodies are detected regularly after infusions of IV Rh immune globulin (RhIG) for the treatment of immune thrombocytopenic purpura (ITP) in D+ patients. STUDY DESIGN AND METHODS: Blood samples from 16 D+ patients with ITP were tested after treatment with IV RhIG for the presence of passively acquired alloantibodies. Similar studies were conducted for three D- patients after injections of IM RhIG for Rh immunoprophyl-axis. Four production lots of IV RhIG and 2 lots of IM RhIG were tested for the presence of alloantibodies. RESULTS: All 16 D+ patients with ITP developed a positive DAT, as well as positive antibody detection test results, after infusions of IV RhIG. All postinfusion plasma samples contained anti-D, as well as one or more additional antibodies, usually anti-C, -E, -G, -V, or -Fy(a). Eluates from patients' RBCs with positive DAT results contained multiple passively acquired alloantibodies. Multiple alloantibodies were detected in samples of different production lots of IV RhIG or IM RhIG. No acute transfusion reactions were observed in five D+ patients with ITP who had been treated with IV RhIG and had been given serologically incompatible D+ RBCs. After injections of IM RhIG, the only passively acquired alloantibody detected was anti-D. CONCLUSION: Plasma samples from D+ patients with ITP treated with IV RhIG regularly contained anti-D and multiple other passively acquired Rh, Duffy, or Kidd system alloantibodies. Postinfusion RBC samples all had positive DAT results with eluates containing anti-D and multiple other Rh, Duffy, or Kidd system antibodies. The consistent detection of multiple passively acquired alloantibodies after IV RhIG, in contrast to the detection of anti-D only after IM RhIG, reflects the immediate effect of the entire (bolus) dose of RhIG by the IV route, the dose for treating ITP that is approximately 10 times the dose for Rh immunoprophylaxis, and the expected serologic incompatibility with recipients' D+ RBCs.     

临床患者血浆中β- 内酰胺类药物＆抗体致输血无效及对供者红细胞亲和力的研究

目的 分析临床患者血浆中β- 内酰胺类药物＆抗体对供者红细胞的亲和力,探究药源性溶血性贫血及输血无效的临床特点。方法 选择2021 年11 月～ 2022 年4 月期间临床送检的4 例有β- 内酰胺类药物用药史的临床患者,检测ABO 与Rh 血型,直接抗球蛋白试验（direct antiglobulin test ,DAT）、不规则抗体鉴定（identification of irregularantibodies,IAT）与β- 内酰胺类药物抗体及效价,对DAT 阳性的患者红细胞进行酸放散并检测放散液中β- 内酰胺类药物抗体。选择ABO 和Rh 同型的供者红细胞与患者血浆在37℃无菌条件下体外致敏,监测β- 内酰胺类药物＆抗体在0 ,24,48 和72 h 与供者红细胞的亲和力,并观察加入补体后的溶血程度。结果 4 例患者血浆中均存在β- 内酰胺类药物抗体,停药前输血无效,停药后输血效果良好。患者1:A 型,RhCCDee,DAT 阳性（3+W）,IAT 阴性,血浆中存在头孢哌酮药物抗体（效价:1∶64）、阿莫西林药物抗体（效价:1∶16）,放散液中存在头孢哌酮药物抗体。患者2:A 型,RhCcDee,DAT 阳性（4+W）,IAT 阴性,血浆中存在头孢哌酮药物抗体（效价:1∶128）,放散液中存在头孢哌酮药物抗体。患者3:A 型,RhCcDee,DAT 阳性（4+）,IAT 阳性,鉴定为抗 E 抗体,血浆中存在阿莫西林药物抗体（效价:1∶16）、亚胺培南药物抗体（效价:1∶64）、头孢哌酮药物抗体（效价:1∶128）,放散液中存在亚胺培南、头孢哌酮药物抗体。患者4:A 型,RhCCDee ,DAT 阳性（1+）,IAT 阳性,鉴定为抗-E.c 抗体,血浆中存在阿莫西林药物抗体（效价:1∶32）,放散液中未检测到药物抗体。4 例患者血浆与供者红细胞体外致敏24 h 后DAT均为阳性,48 和72 h 内逐渐增强,加入补体后均可引起红细胞溶解。结论 头孢哌酮、阿莫西林和亚胺培南三种β-内酰胺类药物＆抗体可迅速结合到供者红细胞表面并随时间延长而增强,有补体存在的条件下可在24 ～ 72 h 内破坏供者红细胞引起溶血,导致输血无效。     

A severe transfusion reaction associated with anti-EnaTS in a patient with an abnormal alpha-like red cell sialoglycoprotein

A 71-year-old woman (Ped) received 3 units of red cells (RBCs), compatible by the indirect antiglobulin test but strongly (4+) incompatible by direct agglutination at 37 degrees C. The next day, her plasma hemoglobin was 1252 mg percent and the direct antiglobulin test (DAT) was weakly positive (IgG and C3). Less than 5 percent of the transfused cells could be detected 48 hours posttransfusion. Her clinical condition deteriorated and renal failure developed. The patient died of pulmonary embolism. Her serum contained a strong (4+) IgM agglutinin and a weakly reactive (microscopically positive) IgG antibody, with anti-EnaTS specificity. EnaFS and EnaTS antigens were severely depressed or absent from the patient's RBCs; the ficin-resistant Ena antigen (EnaFR) appeared to be present. Pretransfusion RBC sialic acid level was 53 +/- 2 percent of normal. The patient's RBC membranes were shown to contain sialoglycoproteins beta and delta by sodium dodecyl sulphate polyacrylamide gel electrophoresis with periodic acid Schiff's base staining, with weak staining of components in the regions corresponding to alpha, alpha 2 and alpha delta. The nature of these components was not identified, but their presence suggested that the patient's RBCs expressed a previously undescribed sialoglycoprotein alpha variant.     

乳腺癌中不同克隆号HER-2抗体IHC与FISH检测结果比较

目的 3种不同克隆号人表皮生长因子受体2(HER-2)抗体的免疫组织化学(IHC)检测结果与荧光原位杂交(FISH)检测结果进行比较,为实验室选择合适的HER-2抗体提供依据。方法选择3种不同克隆号的HER-2抗体,对268例浸润性乳腺癌标本进行检测,并与HER-2基因的荧光原位杂交检测进行比较。结果 HER-2的3种抗体的结果分别为克隆号SP3IHC结果3+66例;克隆号EP3IHC结果3+53例;Polyclonal Her-2抗体IHC结果3+80例,差异有统计学意义(P0.05)。与FISH结果的一致性Kappa值分别为0.76、0.67、0.56。结论 HER-2的IHC检测以选择单克隆抗体优先,建议优先选择克隆号SP3抗体。